Joint Annual Fall Meeting 2002 of GBM and DPGT

Joint Annual Fall Meeting of GBM and DGPT Halle (Saale), September 7-10, 2002

Abstracts Complete list of abstracts Symposia Cellular signalling in animals and plants Complex Molecular Analysis in Clinical Medicine (SG Mol. Medizin) Cytochrome P450 –Multivalent Catalysts for Biotechnology and Pharmacology From nascent chains to protein function From protein transport to organelle development Mol. Mechanisms of development and differentiation of skeletal and cardiac muscles Mol. Structure, function and regulation of xenobiotic metabolising enzymes Molecular medicine and functional genomics New Approaches in Cardiovascular Disease New aspects in GPCR-signalling New molecular approaches to prevent infections Oxidative Stress and Redoxs-Regulation (SG Biochemische Pharmakologie u. Tox.) Pharmacogenomics (Paul-Martini-Stiftung Symposium) Plenary Lectures Proteins involved in apoptosis and transport Proteomics (SG Mol. Zellbiologie, SG Bioanalytik) Transfection with proteins and antisense-RNA _Other (free) topic_ Authors Complete list of authors

Contact GBM e.V. Mörfelder Landstraße 125 60598 Frankfurt am Main phone: 069 / 660567-0 fax: 069 / 660567-22 mail: [email protected] web: www.gbm-online.de DGPT e.V. www.dgpt-online.de Kongressbüro GBM/DGPT Halle 2002 Medizinische Fakultät, Institut für Physiologische Chemie Hollystr.1 D-06097 Halle (Saale) phone: +49-345-557-3840 fax: +49-345-557-3877 mail: [email protected]

Abstracts (complete list) Susana Gariba de Garcia, Gerald Münch, Gabriele Oehme "Carbonyl stress” by methylglyoxal causes mitochondrial dysfunction, ATP depletion and NMDA receptor overstimulation in neurons [poster] Kathrin Fuchs, Diana Kühn, Johannes Schmitt, Joachim Nöller Membrane affinity and pore forming properties of proteins/peptides - Easy and fast by using solid supported lipid bilayers. [poster] Lars-Oliver Klotz, Helmut Sies, Peter Schröder (-)-Epicatechin Is Taken up by Cells and Protects against Peroxynitrite-Induced Oxidation and Nitration. [poster] Cornelia Körting, Alexander Froschauer, Reinhold Hanel, Jean-Nicolas Volff, Anne-Marie Veith DMRT Genes and Sex Determination in the Fish Xiphophorous [poster] Christian Monnerjahn, Ina Schüttmann, Manfred Konrad A designed variant of Herpes virus thymidine kinase selectively phosphorylates the anticancer and antiviral prodrug ganciclovir [oral presentation] Carsten Skarke, Helmut Schmidt, Jürgen Liefhold, Gerd Geisslinger, Jörn Lötsch A mathematical model to predict plasma concentrations of morphine and its active metabolite morphine-6-glucuronide in healthy young persons [poster] Thomas Tuschl, Sayda M. Elbashir, Jens Harborth, Kim Bechert, Mariana Lagos-Quintana, Javier Martinez, Abdullah Yalcin, Agnieszka Patkaniowska, Jutta Meyer, Klaus Weber A new world of tiny RNAs - siRNAs and miRNAs [oral presentation] Nils Hanekop, Susanne Wied, Stefan Petry, Norbert Tennagels, Wendelin Frick, Günter Müller A Receptor for Glycolipid Headgroups Mediates Insulin-mimetic Signaling in Rat Adipocytes [poster] Albert Zlotnik, Anja Muller, Bernhard Homey A role for Chemokine Receptors in Cancer Metastasis? [oral presentation] Sabine Groesch, Irmgard Tegeder, Karin Schilling, Ellen Niederberger, Gerd Geisslinger Activation of c-Jun-N-terminal-kinase by R- and S-flurbiprofen results in cell cycle arrest in human colon carcinoma cells [poster] Kotb Abdelmohsen, Lars-Oliver Klotz, Helmut Sies Activation of EGF receptor-dependent signaling pathways by alkylating and redox-cycling quinones [poster] Hannemann Frank, Bernhardt Rita, Zearo Silvia Activities of CYP11B isoforms from different species. [poster] Svetlana Tsareva, Florian Corvinus, Bernd Wiederanders, Roland Kaufmann, Edith Pfitzner, Richard Moriggl, Karlheinz Friedrich Activities of oncogenic STAT3 in colorectal cancer cells [poster] Andreas Simm, Sherif Daoud, Christian Casselmann, Rolf-Edgar Silber, Reinhard Schinzel Advanced Glycation Endproducts induced cell signalling in cardiac fibroblasts [poster] Markus Kellner, Patrick Baer, Martin Wehling, Ralf Loesel Aldosterone action in humans -new members of the aldosterone signaling pathway- [poster] Carolin Oberle, Astrid Matzke, Silvia Meixner, Ulrich Massing, Harald F. Krug Alkylphosphocholine Analogues induce Apoptosis in Tumour Cells of the Immune System [poster] Sabine Werner, Stefan Kuklinski, Brigitte Schmitz Altered O-GlcNAc level after neuronal differentiation of PC12 cells [poster]

Yvonni Chovolou, Wim Wätjen, Andreas Kampkötter, Regine Kahl ALTERED SENSITIVITY OF TNF-ALPHA OVEREXPRESSING RAT HEPATOMA CELLS AGAINST EXOGENOUS TNF-ALPHA AND OXIDANTS [poster] Jens Wulfänger, Tanja Blosz, Alexander Navarrete Santos, Jürgen Langner, Dagmar Riemann Aminopeptidase N/CD13 is associated with Lubrol rafts [poster] Christoph Hutter, Silke Bergmann, Stefan Burdach, Martin S. Staege Analysis of EWS/Fli-1 induced gene expression in HEK293 cells using DNA-microarrays [poster] Harald F. Krug, Claudia Ball, Silvia Diabate Analysis of oxidative stress due to fly ash and ultrafine particles in macrophages and epithelial cells of the lung [poster] Albert Sickmann, Marion Herrmann, Joachim Klose, Helmut E. Meyer, Heike Schaefer Analysis of Posttranslational Modifications of alpha-A-Crystallin during Aging of the Eye Lens [poster] Klaus Heeg Anti-infective intervention via Toll-like receptors [oral presentation] Kurt Engeland Apoptosis induction and cell cycle regulation at the G2/M-checkpoint by p63, p73 and the tumor suppressor p53 [poster] Daniela Maennel Appropriate TNF production - a decisive factor in sepsis [oral presentation] Michael Spoerner, Guido Steiner, Thomas Prisner, Alfred Wittinghofer, Hans-Robert Kalbitzer Are GTP-analogs really good analogs? [poster] R.T. Sauer, R. Burton, J. Kenniston, S. Joshi, D. Wah, I. Levchenko, R. Grant, J. Flynn, S. Neher, T. Baker ATP-dependent protein degradation and unfolding by ClpXP [oral presentation] Ulrike D. Epple, Henning Barth, Michael Thumm Aut10p, Mai1p and Aut5p shed new lights on the molecular mechanisms of autophagy in S.cerevisiae [oral presentation] Anna Stepczynska, Jakob Troppmair, Thomas Voegel, Monika Poppe, Klaus Schulze-Osthoff, Michael Schwarz Bcl-2 antagonist of cell death (Bad) retains its proapoptotic activity despite proteolytic cleavage by caspase-3 [poster] Otto-Erich Brodde, Rainer Büscher, Kirsten Leineweber, Heike Bruck Beta-adrenoceptor polymorphisms: does altered in vitro function predict in vivo effects? [oral presentation] Ludwig Nißler Binding of flavonoid and anthraquinone derivatives to nucleotide binding domains of ABC transporters. [oral presentation] Claudia Immisch, Alexander Navarrete Santos, Gerald Böhm Binding of RNA to the hyperthermophilic histone-like protein TmHU [poster] Josef H. Wissler, Enno Logemann Biomolecular switch from oxidative stress in inflammation to tissue morphogenesis in healing: Fenton-type OH* radical reactions and isoguanosine in metalloregulated RNA bioaptamers of S100-proteins [poster] Christina Justenhoven, Hiltrud Brauch, Thomas Brüning, Hans-Peter Fischer, Ute Hamann, Volker Harth, Yon Ko, Beate Pesch Breast Cancer and Predictive Factors: Association with Polymorphisms [poster] Ljudmila V. Borissenko, Qinghua Fang, Jens Fey, Thomas Dierks, Kurt von Figura C(alpha)-Formylglycine: a novel protein modification in pro- and eukaryotes [oral presentation] christopher stroh, ajoy samraj, uwe cassens, klaus schulze-osthoff, marek los

Caspase inhibition strongly improves the recovery of cryopreserved hematopoietic and other cells [oral presentation] Harald F. Krug, Silvia Meixner Caspase-10 is the Key Initiator-Caspase involved in TBT-mediated Apoptosis [oral presentation] Romanita Skach, Yon Ko, Jürgen Hescheler, Agapios Sachinidis Catechins with a galloyl group in the 3-position inhibit the PDGF-BB-induced intracellular signal transduction pathway and proliferation of vascular smooth muscle cells [poster] Boudewijn Burgering, Rene Medema, Geert Kops cell cycle and death control: long live Forkheads [oral presentation] Reiner Lammers, Hans-Ulrich Häring, Katja Kapp Cellular transformation mediated by Src kinase: the role of PTPalpha [poster] Matthias Gautschi, Sören Just, Sabine Rospert Chaperons at the Ribosomal Tunnel Exit - First Contacts of a Newly Synthesized Polypeptide [oral presentation] Volker Florian, Peter Knauth, Thomas Schlüter, Susan Schreckenberger Characterisation of Protein Domains Responsible for Recognition of P-Selectin by Sorting Nexin 17 [poster] Joachim Kappler, Volkmar Gieselmann, Frank Dietz, Sebastian Franken Characterization of a new member of the Hepatoma-derived growth factor protein family [poster] Daniel de Graaf, Philip Denner, Luiza Bengtsson, Henning Otto Characterization of Inner Nuclear Membrane Proteins [oral presentation] Anke Hannemann, Marina Bigl, Frank Gaunitz, Klaus Eschrich Characterization of the human platelet-type 6-phosphofructo-1-kinase gene promoter [poster] Matthias Bros, Ute Gödtel-Armbrust, Ulrich Förstermann, Jean-Paul Boissel CHARACTERIZATION OF THE MULTIPLE PROMOTERS OF THE HUMAN NEURONAL NITRIC OXIDE SYNTHASE GENE [oral presentation] XiaoLi Lu, Zhibao Mi, Jeff Mai, Paul Robbins Characterization, Optimization and Application of Peptide-Mediated Protein Transduction [oral presentation] Klaus Heese, Yasuo Nagai, Tohru Sawada Characterizing the signaling pathway of rat p18 amyloid beta (Aß) responsive protein (p18AßrP). [poster] Ivo Zemp Circularly permuted proteins with alternative folding possibilities [poster] F. Westermann, J. S. Wei, M. Ringner, L. H. Saal, F. Berthold, M. Schwab, C Peterson, P Meltzer, J. Khan Classification and diagnostic prediction of pediatric cancers using gene expression profiling and artificial neural networks [oral presentation] Regina Hühn, Beate Liebig, Martin S. Staege, Stefan Burdach Cleavage of EWS/FLI-1 fusion transcripts by Hammerhead Ribozymes [poster] Barth Holger, Klaus Aktories, Christian Wilde Clostridium botulinum C2 toxin as a protein transport system [poster] Klaus Aktories, Christian Wilde Clostridium botulinum C3 exoezyme like ADP-ribosyltransferases [poster] Peter Spangenberg, Nadine Zidek, Marion Grau, Isabel Göhring, Roy Eylenstein, Sven Güttich, Wolfgang Lösche, Stan Heptinstall Comparison of the effects of the P2Y12 ADP-receptor antagonist (AR-C69931) and the GPIIb/IIIa antagonist GR 144053F on platelet-leukocyte conjugate formation. [poster] Stefan Henrich, Robert Huber, Albert Ries, Karlheinz Mann, Klaus Kühn, Rupert Timpl, Gleb P. Bourenkov, Hans D. Bartunik, Wolfram Bode, Manuel E. Than

Crystal structure of the noncollagenous (NC1) domain of human placenta collagen IV: Stabilization via a novel type of covalent Met-Lys cross-link [poster] Ilme Schlichting Crystallographic studies on the reaction mechanism of P450 [oral presentation] Marcel Schmitt, Georg Gellert, Bettina Kirberg, Jost Ludwig, Hella Lichtenberg-Fraté CyGene: Eine neue hefebasierte Methode zur Bestimmung des cyto- und genotoxischen Potentials von Umweltgiften im Wasserbereich. CyGene: A New Yeast Based Method for the Detection of Cyto- and Genotox [poster] Rita Bernhardt Cytochrome P450 dependent steroid hydroxylases and their application in biotechnology and medicine [oral presentation] Birgit Lauterbach, Eduardo Barbosa, Eva Kaergel, Horst Honeck, Jürgen Theuer, Hermann Haller, Friedrich C. Luft, Maik Gollasch, Wolf-Hagen Schunck Cytochrome P450-dependent eicosapentaenoic acid epoxygenation produces novel vasoactive metabolites [oral presentation] Horst Honeck, Dominik N. Müller, Jürgen Theuer, Friedrich C. Luft, Wolf-H. Schunck, Eva Kärgel Cytochrome P450-Dependent Renal Metabolism of Arachidonic Acid in a Rat Model of Angiotensin II induced Hypertension and End-Organ Damage [poster] Eva Borst, Martin Messerle CYTOMEGALOVIRUS-A NEW VECTOR FOR GENE TRANSFER [poster] Elke Deuerling, Günter Kramer, Thomas Rauch, Wolfgang Rist, Sonja Vorderwülbecke, Nenad Ban, Bernd Bukau Cytosolic proteins at birth: linking translation and chaperone assisted protein folding [oral presentation] Juergen Kuhlmann, Alexander Wolf Development of an in vitro system for detection and quantitation of urothelial genotoxicity of tobacco smoke-specific constituents [poster] Rainer Schäfer, Georg Reiser Different regulation of human lung cancer cell proliferation by nucleotides mediated through P2Y receptors [oral presentation] Günther Richter, Karin Hanewinkel, Stefan Burdach Differential analysis of interleukin 2 (IL-2) versus IL-15 induced immediate-early genes: A contribution for the elucidation of the AICD paradoxon? [oral presentation] Claudia Loske, Gerald Münch, Björn Kuhla, Sladjana Dukic-Stefanovic Differential effects of "advanced glycation endproducts" and ß-amyloid peptide on glucose utilization and ATP levels in the neuronal cell line SH-SY5Y [poster] Polin Mahjour, Katja Mühlberg, Andreas Hagendorff, Stefan Dhein, Dietrich Pfeiffer, Aida Salameh Differential regulation of gap junction protein expression in cardiomyocytes [poster] Jürgen Voigt, Stefan Stevanovic, Otfried Marquardt, Ronald Frank Differential subcellular distribution, structure and functions of the 14-3-3 proteins of the unicellular green alga Chlamydomonas reinhardtii [oral presentation] Hans J. Schramm Dimerization inhibitors of HIV-1 protease [poster] Cord-Michael Becker, Bertram Wiedenmann Educational aspects of molecular medicine in basic science and clinical training [oral presentation] M Tharwat Ghoneim, S. H. El-Reweni, A. I. El-Mallah ***, S. H. Hammadi, M. M. Mikhail, A. M. Baraka, S. Kato *, O. El-Sharaki ** Effect of Lacidipine on induced liver fibrosis / cirrhosis. [poster] Stefan Wölfl, Annette Geyer, Larissa Odyvanova, Torsten Kroll, Peter Hortschansky, Manuela Schwalbe, Anke Naumann, Klaus Höffken, Joachim Clement Effect of BMP-2 on the migratory and invasive properties of breast cancer cells [oral presentation]

Meike Strasdat, Slobodan Kovacevic, Martin Paul, Anne Weinstrauch, Robert Real, Hans-Dieter Orzechowski Effect of MAP kinase inhibitors on injury-induced neointima formation in the rat carotid artery [poster] Matthias Eckhardt, Simon Fewou, Volkmar Gieselmann Effect of N-glycan removal on catalytic activity of cerebroside sulfotransferase [poster] Maria Laura Massa, Simone Baltrusch, Sigurd Lenzen, Markus Tiedge Effect of PFK-2 overexpression in insulin-producing cells upon glucokinase activity and glucose metabolism [poster] Armin Kabat, Peter Rösen, Antje Olbrich, Stefan Dhein Effects of vitamin E on endothelial function in diabetes mellitus and hyperglycemia [oral presentation] Lars Boenicke, Regina Pauls, Chu Kang, Holger Kalthoff Efficient Dosage and Time dependent Protein Transduction of Pancreatic Carcinoma Cells using purified VP22-EGFP Fusion Protein [oral presentation] Matthias Kappler, Matthias Kotzsch, Frank Bartel, Susanne Füssel, Chritine Lautenschläger, Peter Würl, Matthias Bache, Hannelore Schmidt, Helge Taubert, Axel Meye Elevated expression level of survivin protein in soft-tissue sarcomas [poster] Klaus Golka, Seidel Thilo, Roetzel Claudia, Geller Frank, Bolt Hermann M, Dietrich Holger, Foth Heidi, Thier Ricarda Enzym polymorphisms and risk factors in bladder cancer patients in Lutherstadt Wittenberg [poster] Thomas Schneider, Gerhard Braus, Gabriele Heinrich, Markus Hartmann, Andrea Pfeil Evolution of Feedback-inhibited (&beta/&alpha)8 Barrel Isoenzymes by Gene Duplication and A Single Mutation [poster] Marietta Kaszkin, Sabine Beck, Gerard Lambeau, Josef Pfeilschifter Exogenous phospholipases A2 mediate gene expression in rat mesangial cells via PPARalpha activation [poster] Gia Anh Bao PHAN, Detlef PIETROWSKI, Christoph KECK EXPRESSION OF A NEW SPLICE VARIANT OF IGFBP-7/MAC25 TUMOR SUPPRESSOR GENE IN HUMAN GRANULOSA CELLS LACKING THE IGFBP MOTIF (GCGCCXXC) [poster] Franziska Bleichert*, J.P. Warnke**, Klaus Eschrich*, Renate Keßler* Expression of ubiquitous 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase in intracranial neoplasms and non-tumorous brain tissue [poster] Ulla Grauschopf Expression, Purification and Characterization of the Inner Membrane Redox Catalyst DsbB from E. coli [oral presentation] Helmut Hanenberg, Detlev Schindler Fanconi anemia: basic molecular biology and clinical implications [oral presentation] Monika Walter, Robert Seckler, Benjamin Heinz Folding Kinetics of Pectate Lyase from Bacillus subtilis [poster] Martin S. Staege, Christoph Hutter, Silke Bergmann, Sabine Körber, Stefan Burdach From genes to antigens: Potential target structures for autologous and allogeneic cytotoxic T cells identified by oligonucleotide arrays [oral presentation] Hans-Dieter Volk, Susanna Prösch From the molecular mechanism of human cytomegalovirus (re)activation to new therapeutical concepts Susanna Prösch and Hans-Dieter Volk Institutes of Virology and Medical Immunology, Humboldt Unive [oral presentation] Reinhild Tenderich, Brigitte Schmitz Function of the PEST sequence of NCAM 140 [poster] André Förster, Jana Gerber, Thomas Juretzek, Gerold Barth, Stephan Mauersberger

FUNCTIONAL EXPRESSION OF HETEROLOGOUS CYTOCHROME P450 IN YARROWIA LIPOLYTICA AND ITS USE IN BIOTRANSFORMATION OF STEROIDS [oral presentation] Christoph Thiele Functional lipidomics [oral presentation] Anne-Claude Gavin, Paola Grandi, Markus Boesche, Roland Krause, Christina Leutwein, Bernhard Kuster, Giulio Superti-Furga Functional organization of the yeast proteome by systemic analysis of protein complexes [oral presentation] Ulrich M. Zanger, Kathrin Klein, Tanja Richter, Thomas E. Mürdter, Ute Hofmann, Thomas Lang, Michel Eichelbaum, Matthias Schwab FUNCTIONAL SIGNIFICANCE OF GENETIC POLYMORPHISMS OF HUMAN CYP2B6 [oral presentation] Wolfgang Hilt, Iris Velten, Martin Ligr, Harish Karnam, Julia Ilyina Functions of the proteasome in cell cycle and apoptosis [oral presentation] Thomas Hartung G-CSF in infection prophylaxis [oral presentation] Stefan Offermanns, Nina Wettschureck, Alexandra Zywietz G-Protein mediated signalling: Studies in conditional Galpha knockouts [oral presentation] Tobias Kanacher, Miguel Pineda, Anita Schultz, Joachim Schultz, Jürgen Linder GAF-domains as novel cAMP-sensor autoactivate a bacterial adenylyl cyclase [poster] Johanna Mansfeld, Eva Petermann, Renate Ulbrich-Hofmann Generation of catalytically active neutral protease from Bacillus stearothermophilus does not require the presence of the propeptide [poster] Monika Stoll Genetic screening approaches for the identification of cardiovascular drug targets [oral presentation] Daniela Besong Genexpression of the human membrane-associated Progesterone Receptor (hmPR) in HepG2 cells [poster] Klaus-Peter Koller Genome wide search for disease genes as new drug targets [oral presentation] Lorenz Trümper, Frank Griesinger, Christa Fonatsch, Astrid Heutelbeck, Detlef Haase, Ernst Hallier, Thomas Schulz GLUTATHIONE S-TRANSFERASE GSTP1 POLYMORPHISMS IN DE NOVO AND THERAPY-INDUCED HEMATOLOGIC MALIGNANCIES [poster] Mike Beck, Kieran Brickley, Miriam Smith, Seema Sharma, Helen Wilkinson, F. Anne Stephenson GRIF-1 - a novel GABAA receptor associated protein [poster] Frank Bartel, Peter Würl, Axel Meye, Matthias Kappler, Matthias Bache, Hannelore Schmidt, Manfred Schönfelder, Helge Taubert Growth reduction of a xenotransplanted human Soft Tissue Sarcoma by MDM2 Antisense Therapy [oral presentation] Wim Wätjen, Yvonni Chovolou, Andreas Kampkötter, Quynh-Hoa Tran-Thi, Regine Kahl, Petra Niering H2O2-INDUCED OXIDATIVE STRESS IN H4IIE CELLS: INFLUENCE OF THE FLAVONOID KAEMPFEROL [poster] Werner Kroll High throughput gene expression profiling to screen for drug effects and toxicity [oral presentation] Heike Schaefer, Yvonne Wagner, Albert Sickmann, Helmut E. Meyer* High throughput mass spectrometry for proteomics [oral presentation] Christian Bamann, Hans-Robert Kalbitzer, Eike Brunner, Astrid Jung High-temperature structure of TmCsp [poster]

Pavel I. Nedvetsky, Alexander Y. Kots, Helmut Müller, Ferid Murad, Harald H.H.W. Schmidt Hsp90 effects on the regulation of soluble guanylyl cyclase [poster] Mario Jakob, Peter Hanner, Ralf Bernd Klösgen Hunting for proteins associated with the delta pH / Tat-pathway in chloroplasts [poster] Bernd Kammerer, Rainer Kahlich, Florence Pojer, Shuming Li, Christoph Gleiter, Lutz Heide Identification of aminocoumarin antibiotics from different Streptomyces species via ESI-MS and LC-MS/MS coupling [poster] Johanna Mansfeld, Renate Ulbrich-Hofmann, Peter Dürrschmidt Identification of an intermediate in the unfolding of neutral protease from Bacillus stearothermophilus by fluorescence spectroscopy [poster] Albert Sickmann, Stefan Lehr, Armin Herkner, Dirk Müller-Wieland, Helmut E. Meyer, Jörg Reinders Identification of the Major Tyrosin Phosphorylation Sites of Human Gab-1 [poster] Kristin Ehrbar, Wolf-Dietrich Hardt Identification of the secretion and translocation domain of Salmonella typhimurium effector protein SopE [poster] Dan Roden Implementing pharmacogenomics to improve drug therapy: where do we stand in 2002 [oral presentation] Joao Pedro Marques, Ingrid Dudeck, Martin Schattat, Ralf Bernd Kloesgen Import and plastid sorting of chimaeric GFP-polypeptides [poster] Paola Pocar, Robert Augustin, Bernd Fischer Induction of apoptotic cell death in bovine cumulus-oocyte complexes after treatment with Aroclor-1254 during in vitro maturation. [poster] Jean-Paul Boissel, Dorothea Ohly, Matthias Bross, Ute Gödtel-Armbrust, Ulrich Förstermann INDUCTION OF THE HUMAN NEURONAL NITRIC OXIDE SYNTHASE EXPRESSION BY EPIDERMAL GROWTH FACTOR IS MEDIATED THROUGH THE JAK/STAT SIGNAL TRANSDUCTION PATHWAY [poster] Kurt von Figura Inherited disorders caused by faulty protein modification in the secretory route [oral presentation] Karin Doods, Karl-Werner Scharmm, Antonius Kettrup INHIBITION OF EROD-ACTIVITY IN RAT HEPATOMA CELLS [poster] V. Krügel Inhibition of LiCl-induced induction of glutamine synthetase in HepG2 cells by flavonoids and anthraquinone derivatives is mediated by CK I [poster] Martin Haßler, Edgar Brändle, Joachim Greven, Jan Henrik Schlattjan Interaction of the organic base cimetidine with the renal basolateral p-aminohippurate (PAH) transport system [poster] Christopher Böhm, Antonia Munck, Wolfgang Hampe Interactionpartners of SorLA [poster] Jörg Bär, Jürgen Kirchberger, Anke Edelmann Interactions between the two types of subunits forming an enzymatically active oligomeric phosphofructokinase-1 from Saccharomyces cerevisiae [poster] Lusine¹ Danielyan, Simone¹ Harsch, Christina¹ Schneider, Gayane² Buniatian, Heiner² Wiesinger, Selim³ Kuçi, Christoph¹ Gleiter Inverse Dynamics of Glial Fibrillary Acidic Protein (GFAP) Expression and Synthesis of Erythropoietin (EPO), EPO-Receptor (EPOR) and Proliferating Cell Nuclear Antigen (PCNA) in Astrocytes [poster] Hubert Zipper, Katrin Lämmle, Christiane Buta, Herwig Brunner, Jürgen Bernhagen, Frank Vitzthum Investigations on the binding of SYBR Green I to double-stranded (ds)DNA [poster] Harald, F. Krug, Tanja Detzel, Siegfried Strack

Involvement of different signalling pathways in cytoskeletal reorganisation during tributyltin (TBT) and H2O2 induced apoptosis [poster] Ina Wittko, Sigrid Saaler-Reinhardt Involvement of La protein in alternative translation processes in neural progenitor cells [poster] Uener Kolukisaoglu, Burkhard Schulz, Bianca Hust, Ulf-Ingo Fluegge, Ralf Bernd Kloesgen, Michael Gutensohn Isolation of an Arabidopsis T-DNA mutant for the chloroplast protein import receptor atToc33 [poster] Peter Gierschik, Monika Gaugler, Ute Dreißigacker, Meike Müller, Klaudia Giehl K-Ras-dependent signal tranduction in human pancreatic carcinoma cells [poster] Kai Simons Lipid rafts in membrane trafficking and cell polarity [oral presentation] Zuebeyde Pamukci, Andreas Koschinski, Eugen Domann, Trinad Chakraborty, Ayub Darji, Dierk Brockmeier, Florian Dreyer, Holger Repp, Jan Birringer Listeriolysin of Listeria monocytogenes forms Ca2+-permeable pores leading to intracellular Ca2+ oscillations in host cells [oral presentation] Melanie Wagner, Reinhard Reents, David Owen, Herbert Waldmann, Alfred Wittinghofer, Jürgen Kuhlmann Localisation of fluorescently labelled Ras protein in the living cell [poster] Arina Kostanian, Heinz Bönisch, Manfred Göthert, Michael Brüss Loss of function of the naturally occurring Arg219Leu variant of the human 5-HT1A receptor [oral presentation] Ralf Baumann, Hans-Uwe Simon MACROPHAGE MIGRATION INHIBITORY FACTOR (MIF) IS A SURVIVAL FACTOR FOR NEUTROPHILS [poster] Andreas Humeny MALDI-TOF-MS Based Analysis of Disease-related Gene Variants [oral presentation] F. Peter Guengerich Mechanisms of Oxidations of Drugs and Carcinogens Catalyzed by Human P450s [oral presentation] Horst Honeck, Wolf-Hagen Schunck, Eduardo Barbosa Sicard Metabolism of eicosapentaenoic acid by CYP4A and CYP2C enzymes [poster] Susanne Ammon, Peter Mayer, Volker Höllt Microarray analysis fo genes expressed in the frontal cortex of rats chronically treated with morphine [poster] Michael Höcker, Anna Walduck, Christian Wunder, Stefan Jüttner, Bertram Wiedenmann, Michael Naumann, Thomas F. Meyer Microarray analysis of mucosal gene expression profiles during gastric H. pylori infection [oral presentation] Nils Wiedemann, Kaye Truscott, Peter Rehling, Chris Meisinger, Nikolaus Pfanner Mitochondrial translocases for precursor proteins [oral presentation] Ellen Niederberger, Achim Schmidtko, Jeffrey D. Rothstein, Gerd Geisslinger, Irmgard Tegeder Modulation of spinal nociceptive processing through the glutamate transporter GLT-1 [poster] Hannah Schröder-Borm, Regine Willumeit, Jörg Andrä Molecular basis for membrane selectivity of a potent peptide antibiotic derived from NK-lysin [oral presentation] Peter Rehling Molecular Dissection of the Protein Import Complexes of the Inner Membrane of Yeast Mitochondria [oral presentation] Franz-Josef Neumann Molecular mechanisms of restenosis [oral presentation]

Jürgen Radons, Reinhold Altmann, Alexander Botzki, Stefan Dove, Werner Falk Molecular modeling as a tool to identify putative interacting sites in the IL-1 receptor complex [poster] Heinrich Sticht Molecular modelling of HIV regulatory proteins as targets of viral infectivity [oral presentation] Leopold Flohé Molecular tools to design trypanocidal drugs [poster] Ekkehardt Stehfest, Abdel Torky, Heidi Foth MRP expression and modulation of/by glutathione in human lung cells [poster] Martin Augsten, Ignacio Rubio, Karlheinz Friedrich Multivalent Scavengers of Oncogenic Ras: A novel approach to tracking and therapeutical inhibition of Ras activity [poster] Michael Rudnicki, Patrick Seale, Atsushi Asakura, Anna Polesskaya, Anthony Scime Myogenic Specification of Adult Stem Cells in Skeletal Muscle [oral presentation] Wiebke Scherf, Günther Richter, Jana Bergmann, Stefan Burdach, Gesine Hansen Nasal DNA vaccination with an Interleukin-10 cDNA vector in a murine asthma modell [poster] Marc Parmentier, Masato Kotani, Valérie Wittamer, David Communi, Isabelle Migeotte, Jalal Vakili, Michel Detheux, Stéphane Brézillon, Emmanuel Le Poul, Jean-Denis Franssen Natural ligand identification and functional characterization of orphan G protein-coupled receptors [oral presentation] Andreas Busch NHE-1 inhibitors: From acute cardioprotection to heart failure [oral presentation] Jochen Frenzel, Jan Richter, Klaus Eschrich Nitrogen oxide-induced cell death in rabbit cultured Müller cells is modulated by glucose [poster] Andreas Plückthun Novel proteins from combinatorial and evolutionary approaches [oral presentation] Mathias Gautel Novel signalling pathways in striated muscles [oral presentation] Jörg Kahle, Marc Bäuerle, Matthias Baake, Detlef Doenecke, Werner Albig Nuclear import of histones [poster] Martin Göttlicher Nuclear receptors and their role in the regulation of xenobiotic metabolism [oral presentation] Michel Bouvier Oligomeric assembly of G protein-coupled receptors: Role in receptor trafficking and function [oral presentation] Navarrete Santos Alexander, Immisch Claudia, Reinke Claudia, Schaupp Michael, Böhm Gerald On the use of a histone-like protein for drug delivery using doxorubicin as a model system for tumor therapy [poster] Frank Bier On-chip synthesis of extended, long-chain DNA by PCR Xenia Marschan, Dennie Andresen, Markus von Nickisch-Rosenegk, Frank F. Bier [poster] Susanne Brakmann, Sylvia Löbermann, Petra Nieckchen Optimal enzymes for single molecule sequencing [poster] Gudrun Horn, Daniel Weinfurtner, Roland Hofweber, Robert Macek, Robert Gail, Till Maurer, Hans Robert Kalbitzer Optimization of intein based isotope labeling methods for NMR investigations of proteins [poster] Karsten Hemmrich, Christoph V. Suschek, Guido Lerzynski, Victoria Kolb-Bachofen OPTIMIZED ANTISENSE OLIGONUCLEOTIDE-MEDIATED INHIBITION OF THE INDUCIBLE NITRIC OXIDE SYNTHASE CO-SUPPRESSES NO-MEDIATED STRESS RESPONSE IN ENDOTHELIUM [poster]

Lars-Oliver Klotz Oxidant-induced signaling: from intra- to intercellular communication [oral presentation] Harald F. Krug, Silvia Diabaté, Katja Völkel Oxidative stress in rat alveolar epithelial cells and co-cultures upon treatment with fly ash [oral presentation] Jacques POUYSSEGUR, Veronique VOLMAT, Gilles PAGES, Philippe LENORMAND p42/p44 MAP kinase (Erk) - Control of spatio-temporal activity [oral presentation] Nishith Gupta, Katrin Lehmann, Otmar Asperger P450non System from Acinetobacter sp. EB104: A Unique Bacterial Alkane Hydroxylase [oral presentation] Kent Söe, Hella Hartmann, Holger Stephan, Bernhard Schlott, Tinna Stevnsner*, Frank Grosse p53 is involved in the repair of human topoisomerase I cleavage complexes [poster] Agapios Sachinidis, Jürgen Hescheler PDGF-BB induces selective differentiation of mouse embryonic stem cells to cardiac cells [oral presentation] Robert Seckler, Monika Walter Pectate Lyase from Bacillus subtilis (BsPel): Mutations in the asparagine ladder cause a temperature-sensitive folding phenotype [poster] Monika Walter Pectate Lyase from Bacillus subtilis (BsPel): Mutations in the asparagine ladder cause a temperature-sensitive folding phenotype [poster] Julian Arbuckle Pharmacogenetic based clinical trials to improve drug development [oral presentation] Dietmar Pfeifer, Martina Kaufmann, Bernd Fiebich, Olivia Spleiss Pharmacogenetic profiling with Biochips: an innovative concept to improve the efficiency of clinical trials [poster] Ivar Roots, Julia Kirchheiner, Jürgen Brockmöller, Christian Meisel, Kersti Oselin, Thomas Gerloff Pharmacogenetic studies on drug-metabolizing cytochromes P450 and possibilities for individual drug therapy [oral presentation] Heyo K. Kroemer, Werner Siegmund, Ingolf Cascorbi Pharmacogenetics in the Cardiovascular Field [oral presentation] Susanne Haufe, Ines Schäffner, Renate Ulbrich-Hofmann Phospholipase D from cabbage: Calcium ions essential for activity destabilize the protein conformation [poster] Suisheng Zhang, Carsten Köhler, Frank Grosse PHYSICAL INTERACTION BETWEEN NUCLEAR DNA HELICASE II AND WRN HELICASE STIMULATES WRN´S EXONUCLEASE ACTIVITY [poster] Sabine Körber, Sandra Barth, Ulrich Schmidt, Christiane Jäger, Constanze Günther, Christoph Parthier, Gerald Böhm Polyomavirus-like particles as a system for gene therapy and drug delivery [poster] Walter-Vesely Seb. Meister Polyvinylnucleobases: Synthesis, characterization and applications in nanotechnology [poster] Wim Wätjen, Yvonni Chovolou, Petra Niering, Andreas Kampkötter, Gudrun Aussel, Quynh-Hoa Tran-Thi, Regine Kahl PRO- AND ANTIAPOPTOTIC EFFECTS OF QUERCETIN AND FISETIN IN H4IIE-CELLS: IMPLICATION OF OXIDATIVE STRESS [oral presentation] Calin-Aurel Dragan, Rita Bernhardt, Matthias Bureik Production of steroid hormones by fission yeast cells expressing human mitochondrial cytochromes P450 [poster] Sladjana Dukic-Stefanovic, Jovana Gasic-Milenkovic, Winnie Deuther-Conrad, Gerald Münch

Proinflammatory signal transduction pathways in N11 mouse microglial cell line activated by "Advanced Glycation Endproducts" [poster] Sandra Loeppen, Daniela Schneider, Frank Gaunitz, Rolf Gebhardt, Albrecht Buchmann, Michael Schwarz Promotion of hepatocarcinogenesis is mediated by overexpression of glutamine synthetase in beta-catenine-mutated mouse liver tumors [poster] Johannes Herrmann, Tom Lutz, Stephan Meier, Marc Preuss, Gregor Szyrach, Frank Baumann Protein insertion into the inner membrane of mitochondria [oral presentation] M. Cristina Cardoso, Wolfgang Derer, Hariharan P. Easwaran, Heinrich Leonhardt Protein therapy: Applications in tissue regeneration and stem cell research [oral presentation] Tom Rapoport Protein Transport In and Out of the ER [oral presentation] Dietrich Keppler, Yunhai Cui, Jörg König, Anne T. Nies Proteins mediating vectorial transport across plasma membrane domains [oral presentation] Ulrike Sattler, Ingrid Koziollek-Drechsler, Danuta Dormann, Christina Zechel Putative role of the nuclear factor NCNF in neuronal differentiation [poster] Thomas Langer Quality control of membrane proteins in mitochondria [oral presentation] Bernd Moritz, Gudrun Franke, Martin Siemann, Matthias Reuss, Helmut E. Meyer Quantitative Proteomics with Escherichia coli [poster] Kerstin Schnurr, Dagmar Kupper, Antje Banning, Cordula Müller, Matilde Maiorino, Gaby Böl, Regina Brigelius-Flohé Redox events in IL-1 signaling [oral presentation] Arne Holmgren Redox regulation by thioredoxin and glutaredoxin systems [oral presentation] Margarita Fuhrmann, Kurt Racké, Folker Wenzel Reduced NO synthesis during halothane exposure in stimulated rat alveolar macrophages [poster] Marcel Eugster, Ulla Grauschopf Refolding and Characterization of the N-terminal extracellular fragment of the Pituitary Adenylate-Cyclase Activating Peptide Receptor [poster] Frank Fürst, Robert Seckler Refolding from Fragments: The case of the proteolytically processed Pea Lectin [poster] Georg Reiser, Mikhail Strokin Regulation of docosahexaenoic and arachidonic acid release in rat astrocytes [poster] Thomas Braun Regulation of muscle cell development and regeneration by growth factors and bHLH proteins [poster] Bernd Epe Relevance of endogenous oxidative DNA damage: lessons from repair-deficient mice [oral presentation] Jürgen Alves, Dorothee von Witzendorff, Minh-Cam Ha-Thi, Susanne von Pall de Tolna, Gabriele Grabowski Restriction endonuclease EcoRI - fusions for a change of specificity [poster] Frank Czubayko, Achim Aigner Ribozyme-PEI-complexes as ready-to-use reagents for efficient down-regulation of gene expression [oral presentation] Josef H. Wissler RNA as a cytokine [ribokine]: Metalloregulated extracellular eRNA and bioaptamers, the uncovered missing links for cellular communication and signal transduction in an anticipated RNA world of today. [poster]

Jovana Gasic-Milenkovic, Sladjana Dukic-Stefanovic, Winnie Deuther-Conrad, Gerald Münch ß-amyloid peptide potentiates inflammatory responses induced by lipopolysaccharide, interferon-gamma and "advanced glycation endproducts" in a mouse microglia cell line [poster] Anton Vichalkovski, Karin Kirschner, Kurt Baltensperger, Hartmut Porzig SDF-1 and thrombin signaling in human hematopoietic progenitor cells is regulated by cytokines [oral presentation] Ernst Petzinger, Kerstin Lischka, Dieter Starke Secretion of intact oligonucleotides by mrp2 into bile [oral presentation] Antonios Kyriakopoulos, Barbara Hoppe, Alexandra Graebert, Marcus Kühbacher, Dietrich Behne Selenoproteins in the perinuclear structures of rat kidney Dedt. Molecular Trace Element research in the Life Sciences Hahn- Meitner-Istitut, Glienicker Str.100, 14109 Berlin [poster] Maria Ludewig*, Carsten Schwarz+, Otmar Asperger*, Svante Pääbo+, Ulrich Hahn* Sequence Analysis of an Acinetobacter Plasmid Carrying the Genes of a P450-Dependent Alkane Monooxygenase [oral presentation] Alexandra Koch, Annalisa Mancini, Regina Wilms, Teruko Tamura SH2-domain containing inositol 5-phosphatase (SHIP)-1 is a potential regulator of cytoskeletal rearrangements in NGF induced neurite outgrowth [poster] Thorsten Nürnberger, Dierk Scheel Signal perception and transduction in the plant immune response [oral presentation] Julia Lintzel, Inga Franke, Wolfgang Hampe Signal transduction by SorLA, a member of the LDL-receptor family [poster] Manfred Frasch, Hsiu-Hsiang Lee, Patrick Lo, Ingolf Reim, Hideyuki Nagaso SIGNALING AND TRANSCRIPTIONAL CONTROL OF VISCERAL MUSCLE AND HEART DEVELOPMENT IN DROSOPHILA [oral presentation] Ulrich Laufs Significance of cholesterol-independent effects of statins [oral presentation] Claudia von Montfort, Lars-Oliver Klotz Singlet Molecular Oxygen Reversibly Inhibits Tyrosine Phosphatases [poster] Ralph-Peter Elsner Solution structure of the Ras binding domain of AF6 [poster] Alexander Freiberg, Jan Lengefeld, Matthias Machner, Wolfgang Pfeil, Dirk Heinz, Robert Seckler Stability and Folding of Internalin B - A Biophysical Characterization [poster] Matthias Gaestel Stress-signalling downstream to p38 MAPK: Essential role of MAPKAP kinase 2 (MK2) [oral presentation] Stephan Ort, Manfred Konrad Structural and functional analysis of deoxyribonucleoside kinases dCK and dGK: design of novel mutant enzymes for therapeutic applications. [poster] Ingo Paarmann, Manfred Konrad Structural requirements for calmodulin binding to membrane-associated guaylate kinases [poster] Colin Robinson Structure and mechanism of the twin-arginine translocase [oral presentation] Oliver Ohlenschläger, Jens Wöhnert, Enrico Bucci, Ramadurai Ramachandran, Roland Zell, Matthias Görlach Structure of an RNA Involved in Enteroviral Replication [poster] Christoph Kluck, Holger Patzelt, Bernd Bukau, Matthias Mayer Structure-function analysis of HscC, the E. coli member of a novel subfamily of specialized Hsp70 chaperones [poster] Michael Arand

Structure-function relationships of epoxide hydrolases [oral presentation] Christian Lücke, André Richardt*, Heinz Rüterjans, Vicky Katsemi Study on the function and kinetics of diisopropylfluorophosphatase [poster] Cordula Enenkel Targeting mechanism of proteasomes to nuclear / ER membranes and into the nucleus of yeast [oral presentation] Joao Pedro Marques, Ingrid Dudeck, Martin Schattat, Ralf Bernd Klösgen Targeting of chimaeric GFP-polypeptide fusions within chloroplasts [poster] Heinrich Jung, Maike Rochon, Corinna Weber-Sparenberg Targeting of proteins to the flagellar type III secretion system of Escherichia coli [poster] Peter Heimann, Harald Jockusch, Christiane Wiegand Temperature-sensitive TMV Coat Proteins as Pathogens in Animal Cells [poster] Volkmar Gieselmann, Uwe Rauch, Karl Schilling, Penka Pesheva, Sebastian Franken, Stephan L. Baader, Joachim Kappler Tenascins are associated with lipid rafts isolated from mouse brain [poster] heiko funke-kaiser, julia lemmer, christian von langsdorff, alexander thomas, slobodan d. Kovacevic, meike strasdat, frank zollmann, martin paul, hans-dieter orzechowski The cardiac transcription factor Nkx2-5 regulates the alternative promoters of the endothelin-converting enzyme-1 gene in H9c2 cardiomyoblasts [oral presentation] Gabi Bauer-Manz, Andreas Kuhn, Lambertus van den Berg, Justyna Serek The E.coli membrane insertase YidC: A Chaperone for Protein Folding into the Membrane [poster] Leopold Flohé The element of the moon moonlighting for fertility [poster] Heide Schmid, Thai-Hoa Nguyen-Xuan, Minh-Huy Tran, Gerold Schwarz, Hubert Kalbacher, Heinrich Wiesinger The endosomal/lysosomal distribution of cathepsins (cat) B, L and S and N-acetyl-ß-D-glucosaminidase (NAG) is differentially modulated in murine cell lines of microglial and monocytic origin [poster] Astrid Schön, Olaf Gimple, Christian Heubeck The evolution of RNase P in eukaryotic organelles [poster] Detlef PIETROWSKI, Gia Anh Bao PHAN, Paulina WITT, Christoph KECK THE EXPRESSION OF ANGIOGENIC GROWTH FACTORS IN HUMAN GRANULOSA CELLS ARE CONTROLLED BY HUMAN CHORION GONADOTROPIN. [poster] Christina Deschermeier, Georg Mayr, Werner Fanick, Marcus Nalaskowski The human inositol phosphate multikinase (IPMK) is targeted to the nucleus of mammalian cells by a basic cluster in its C-terminus [poster] Armin Huber, Reinhard Paulsen, Claudia Franz The INAD signaling complex of Drosophila photoreceptor: Recombinant Expression and Assembly in a Cell Culture System [poster] Anne Navarrete Santos, Sarah Tonack, Michaela Kirstein, Bernd Fischer The insulin receptor and the insulin-dependent glucose transporter isoform 4 are expressed in rabbit preimplantation embryos [oral presentation] Jens Wulfaenger, Alexander Navarrete Santos, Tanja Blosz, Juergen Langner, Dagmar Riemann The intracellular N-terminus of aminopeptidase N/CD13 affects association with caveolae [poster] Jeroen Buters The knockout approach to understand the function of xenobiotic metabolising enzymes [oral presentation] Ralf Bernd Klösgen, Bo Hou The mechanism of deltapH/TAT-dependent protein translocation across the thylakoid membrane [oral presentation]

Hassan Dihazi, Klaus Eschrich The Mechanism of regulation of yeast 6-phosphofructo-2-kinase by glucose and hypertonic stress [poster] Brigitte Schmitz, Julia Kellersmann The O-GlcNAc modification of proteins interferes with signaling by PKA and CDK5 dependent phosphorylation in neurons [poster] Andreas Strub, Karin Röttgers, Wolfgang Voos The peptide-binding domain determines the interaction of mitochondrial Hsp70s with the essential partner protein Tim44 [poster] Doris Kloor1, Angelika Lüdtke1, Stanka Stoeva2, Hartmut Osswald1 The ratio of the cofactor NAD +/NADH of S-adenosylhomocysteine hydrolase controls the adenosine binding sites. [oral presentation] Jörg Tatzelt, Konstanze F. Winklhofer The role of co- and posttranslational modifications in the propagation of scrapie-prions [poster] Seigo Izumo The role of Csx/Nkx 2.5 homeobox gene in cardiac development and congenital heart disease. [oral presentation] Garret A. Fitzgerald The role of cyclooxygenases in the cardiovascular system [oral presentation] Simone Diestel, Claudia Müller, Marie-Therese Fergen, Brigitte Schmitz The role of NCAM phosphorylation on NCAM mediated signal transduction pathways [poster] Martina Schultheis, Brigitte Schmitz The role of serine-phosphorylation of the cell adhesion molecule L1 in basal neurite outgrowth [poster] Stefanie Barbirz The tailspike from E.Coli H bacteriophage HK620: Another Parallel beta-Helix Protein? [poster] Peter Rehling, Peter Kovermann, Richard Wagner, Nikolaus Pfanner, Kaye, N. Truscott, Katrin Brandner The TIM22 translocase mediates insertion of carrier precursors into the mitochondrial inner membrane via a two-step process [poster] Steffen Liedtke The Use of Multidimensional Capillary LC in Proteomics [oral presentation] Simone Fulda, Klaus-Michael Debatin Therapeutic modulation of death receptor- or drug-mediated apoptosis [oral presentation] Tanja Richter, Kathrin Klein, Thomas E. Mürdter, Michel Eichelbaum, Matthias Schwab, Ulrich M. Zanger ThioTEPA and Clopidogrel are specific mechanism-based inhibitors of human CYP2B6 [poster] Thomas Dandekar Tools of bioinformatics: Comparative genome analysis and pathway reconstruction [oral presentation] Franz Oesch Toxicology from experimental systems to man: drug metabolising enzymes as key determinants [oral presentation] S. Breidert, R. Jacob, A. Ngezahajo, H.-A. Kolb, H. Naim Trafficking pathways of connexin49-GFP in living mammalian cells [poster] Markus von Nickisch-Rosenegk, Jenny Steffen, Frank F. Bier Transcription of Reporter Genes with Immobilized Templates [poster] Michael Metzlaff Transgene- and virus-mediated gene silencing - What plants can teach us! [oral presentation] Dirk H. Ostareck

Translational activation from DICE-mediated mRNA silencing by the Src tyrosine kinase [oral presentation] Sabine Molik, Ralf Bernd Klösgen Transport and assembly of the Rieske Fe/S protein: What happens in the stromal space? [poster] Joerg W. Bartsch, Dirk Wildeboer, Uwe Schlomann Tumor necrosis factor alpha signalling in reactive glia cells [poster] Albert Sickmann, Helmut E. Meyer, Yvonne Wagner Two-dimensional chromatography in protein analysis [poster] Andrè Reis Understanding the genetic basis of common human diseases and its potential for new concepts in drug therapy [oral presentation] Gunter Fischer Unfolding the Chemistry of Protein Folding [oral presentation] Annette Deul, Susanne Stevens, Uta Sander, Christian Kirchhoff, Joern Jungmann, Thomas Hesterkamp Use of cytochrome P450 isoenzyme assays in the hit-to-lead process of drug discovery [poster] Stefan Welte, Günter Müller, Matthias Dreyer, Stefan Petry, Norbert Tennagels, Robert Schwenk Use of PTP1B mutants for selective inhibitor screening [poster] Andreas Bichet, Frank Hannemann, Rita Bernhardt Using the yeast two-hybrid system as in vivo screening-system for interactions between adrenodoxin and its redoxpartners [poster] Paul Pfluger, Marc Birringer, Regina Brigelius-Flohé, Ralph Rühl, Nico Landes vitamin E [oral presentation] Peter Kühl Who discovered the Michaelis-Menten hyperbola? [poster] Kerstin Reichert, Matthias Rödel, Ralph Menzel Xenobiotically induced cytochrome P450 gene expression in Caenorhabditis elegans [poster] Ross Dalbey YidC, a novel evolutionarily conserved protein, mediates membrane protein assembly in bacteria [oral presentation]

Symposium: Cellular signalling in animals and plants Martin Augsten, Ignacio Rubio, Karlheinz Friedrich Multivalent Scavengers of Oncogenic Ras: A novel approach to tracking and therapeutical inhibition of Ras activity Joerg W. Bartsch, Dirk Wildeboer, Uwe Schlomann Tumor necrosis factor alpha signalling in reactive glia cells Christopher Böhm, Antonia Munck, Wolfgang Hampe Interactionpartners of SorLA Jean-Paul Boissel, Dorothea Ohly, Matthias Bross, Ute Gödtel-Armbrust, Ulrich Förstermann INDUCTION OF THE HUMAN NEURONAL NITRIC OXIDE SYNTHASE EXPRESSION BY EPIDERMAL GROWTH FACTOR IS MEDIATED THROUGH THE JAK/STAT SIGNAL TRANSDUCTION PATHWAY Boudewijn Burgering, Rene Medema, Geert Kops cell cycle and death control: long live Forkheads Simone Diestel, Claudia Müller, Marie-Therese Fergen, Brigitte Schmitz The role of NCAM phosphorylation on NCAM mediated signal transduction pathways Kurt Engeland Apoptosis induction and cell cycle regulation at the G2/M-checkpoint by p63, p73 and the tumor suppressor p53 Hassan Dihazi, Klaus Eschrich The Mechanism of regulation of yeast 6-phosphofructo-2-kinase by glucose and hypertonic stress Joachim Kappler, Volkmar Gieselmann, Frank Dietz, Sebastian Franken Characterization of a new member of the Hepatoma-derived growth factor protein family Armin Huber, Reinhard Paulsen, Claudia Franz The INAD signaling complex of Drosophila photoreceptor: Recombinant Expression and Assembly in a Cell Culture System Matthias Gaestel Stress-signalling downstream to p38 MAPK: Essential role of MAPKAP kinase 2 (MK2) M Tharwat Ghoneim, S. H. El-Reweni, A. I. El-Mallah ***, S. H. Hammadi, M. M. Mikhail, A. M. Baraka, S. Kato *, O. El-Sharaki ** Effect of Lacidipine on induced liver fibrosis / cirrhosis. Julia Lintzel, Inga Franke, Wolfgang Hampe Signal transduction by SorLA, a member of the LDL-receptor family Volkmar Gieselmann, Uwe Rauch, Karl Schilling, Penka Pesheva, Sebastian Franken, Stephan L. Baader, Joachim Kappler Tenascins are associated with lipid rafts isolated from mouse brain Marietta Kaszkin, Sabine Beck, Gerard Lambeau, Josef Pfeilschifter Exogenous phospholipases A2 mediate gene expression in rat mesangial cells via PPARalpha activation Brigitte Schmitz, Julia Kellersmann The O-GlcNAc modification of proteins interferes with signaling by PKA and CDK5 dependent phosphorylation in neurons Alexandra Koch, Annalisa Mancini, Regina Wilms, Teruko Tamura SH2-domain containing inositol 5-phosphatase (SHIP)-1 is a potential regulator of cytoskeletal rearrangements in NGF induced neurite outgrowth Christina Deschermeier, Georg Mayr, Werner Fanick, Marcus Nalaskowski

The human inositol phosphate multikinase (IPMK) is targeted to the nucleus of mammalian cells by a basic cluster in its C-terminus Thorsten Nürnberger, Dierk Scheel Signal perception and transduction in the plant immune response Dirk H. Ostareck Translational activation from DICE-mediated mRNA silencing by the Src tyrosine kinase Jacques POUYSSEGUR, Veronique VOLMAT, Gilles PAGES, Philippe LENORMAND p42/p44 MAP kinase (Erk) - Control of spatio-temporal activity Jürgen Radons, Reinhold Altmann, Alexander Botzki, Stefan Dove, Werner Falk Molecular modeling as a tool to identify putative interacting sites in the IL-1 receptor complex Romanita Skach, Yon Ko, Jürgen Hescheler, Agapios Sachinidis Catechins with a galloyl group in the 3-position inhibit the PDGF-BB-induced intracellular signal transduction pathway and proliferation of vascular smooth muscle cells Martina Schultheis, Brigitte Schmitz The role of serine-phosphorylation of the cell adhesion molecule L1 in basal neurite outgrowth Georg Reiser, Mikhail Strokin Regulation of docosahexaenoic and arachidonic acid release in rat astrocytes Svetlana Tsareva, Florian Corvinus, Bernd Wiederanders, Roland Kaufmann, Edith Pfitzner, Richard Moriggl, Karlheinz Friedrich Activities of oncogenic STAT3 in colorectal cancer cells Jürgen Voigt, Stefan Stevanovic, Otfried Marquardt, Ronald Frank Differential subcellular distribution, structure and functions of the 14-3-3 proteins of the unicellular green alga Chlamydomonas reinhardtii Peter Heimann, Harald Jockusch, Christiane Wiegand Temperature-sensitive TMV Coat Proteins as Pathogens in Animal Cells Josef H. Wissler RNA as a cytokine [ribokine]: Metalloregulated extracellular eRNA and bioaptamers, the uncovered missing links for cellular communication and signal transduction in an anticipated RNA world of today.

Symposium: Complex Molecular Analysis in Clinical Medicine (SG Mol. Medizin) Cord-Michael Becker, Bertram Wiedenmann Educational aspects of molecular medicine in basic science and clinical training Thomas Dandekar Tools of bioinformatics: Comparative genome analysis and pathway reconstruction Michael Höcker, Anna Walduck, Christian Wunder, Stefan Jüttner, Bertram Wiedenmann, Michael Naumann, Thomas F. Meyer Microarray analysis of mucosal gene expression profiles during gastric H. pylori infection Andreas Humeny MALDI-TOF-MS Based Analysis of Disease-related Gene Variants Heinrich Sticht Molecular modelling of HIV regulatory proteins as targets of viral infectivity Christoph Thiele Functional lipidomics

Symposium: Cytochrome P450 –Multivalent Catalysts for Biotechnology and Pharmacology Nishith Gupta, Katrin Lehmann, Otmar Asperger P450non System from Acinetobacter sp. EB104: A Unique Bacterial Alkane Hydroxylase Maria Ludewig*, Carsten Schwarz+, Otmar Asperger*, Svante Pääbo+, Ulrich Hahn* Sequence Analysis of an Acinetobacter Plasmid Carrying the Genes of a P450-Dependent Alkane Monooxygenase Horst Honeck, Wolf-Hagen Schunck, Eduardo Barbosa Sicard Metabolism of eicosapentaenoic acid by CYP4A and CYP2C enzymes Rita Bernhardt Cytochrome P450 dependent steroid hydroxylases and their application in biotechnology and medicine Andreas Bichet, Frank Hannemann, Rita Bernhardt Using the yeast two-hybrid system as in vivo screening-system for interactions between adrenodoxin and its redoxpartners Calin-Aurel Dragan, Rita Bernhardt, Matthias Bureik Production of steroid hormones by fission yeast cells expressing human mitochondrial cytochromes P450 F. Peter Guengerich Mechanisms of Oxidations of Drugs and Carcinogens Catalyzed by Human P450s Annette Deul, Susanne Stevens, Uta Sander, Christian Kirchhoff, Joern Jungmann, Thomas Hesterkamp Use of cytochrome P450 isoenzyme assays in the hit-to-lead process of drug discovery Horst Honeck, Dominik N. Müller, Jürgen Theuer, Friedrich C. Luft, Wolf-H. Schunck, Eva Kärgel Cytochrome P450-Dependent Renal Metabolism of Arachidonic Acid in a Rat Model of Angiotensin II induced Hypertension and End-Organ Damage André Förster, Jana Gerber, Thomas Juretzek, Gerold Barth, Stephan Mauersberger FUNCTIONAL EXPRESSION OF HETEROLOGOUS CYTOCHROME P450 IN YARROWIA LIPOLYTICA AND ITS USE IN BIOTRANSFORMATION OF STEROIDS Kerstin Reichert, Matthias Rödel, Ralph Menzel Xenobiotically induced cytochrome P450 gene expression in Caenorhabditis elegans Dietmar Pfeifer, Martina Kaufmann, Bernd Fiebich, Olivia Spleiss Pharmacogenetic profiling with Biochips: an innovative concept to improve the efficiency of clinical trials Tanja Richter, Kathrin Klein, Thomas E. Mürdter, Michel Eichelbaum, Matthias Schwab, Ulrich M. Zanger ThioTEPA and Clopidogrel are specific mechanism-based inhibitors of human CYP2B6 Ivar Roots, Julia Kirchheiner, Jürgen Brockmöller, Christian Meisel, Kersti Oselin, Thomas Gerloff Pharmacogenetic studies on drug-metabolizing cytochromes P450 and possibilities for individual drug therapy Ilme Schlichting Crystallographic studies on the reaction mechanism of P450 Birgit Lauterbach, Eduardo Barbosa, Eva Kaergel, Horst Honeck, Jürgen Theuer, Hermann Haller, Friedrich C. Luft, Maik Gollasch, Wolf-Hagen Schunck Cytochrome P450-dependent eicosapentaenoic acid epoxygenation produces novel vasoactive metabolites

Hannemann Frank, Bernhardt Rita, Zearo Silvia Activities of CYP11B isoforms from different species. Ulrich M. Zanger, Kathrin Klein, Tanja Richter, Thomas E. Mürdter, Ute Hofmann, Thomas Lang, Michel Eichelbaum, Matthias Schwab FUNCTIONAL SIGNIFICANCE OF GENETIC POLYMORPHISMS OF HUMAN CYP2B6

Symposium: From nascent chains to protein function Elke Deuerling, Günter Kramer, Thomas Rauch, Wolfgang Rist, Sonja Vorderwülbecke, Nenad Ban, Bernd Bukau Cytosolic proteins at birth: linking translation and chaperone assisted protein folding Marcel Eugster, Ulla Grauschopf Refolding and Characterization of the N-terminal extracellular fragment of the Pituitary Adenylate-Cyclase Activating Peptide Receptor Frank Fürst, Robert Seckler Refolding from Fragments: The case of the proteolytically processed Pea Lectin Ulla Grauschopf Expression, Purification and Characterization of the Inner Membrane Redox Catalyst DsbB from E. coli Susanne Haufe, Ines Schäffner, Renate Ulbrich-Hofmann Phospholipase D from cabbage: Calcium ions essential for activity destabilize the protein conformation Christian Bamann, Hans-Robert Kalbitzer, Eike Brunner, Astrid Jung High-temperature structure of TmCsp Christian Lücke, André Richardt*, Heinz Rüterjans, Vicky Katsemi Study on the function and kinetics of diisopropylfluorophosphatase Thomas Langer Quality control of membrane proteins in mitochondria Christoph Kluck, Holger Patzelt, Bernd Bukau, Matthias Mayer Structure-function analysis of HscC, the E. coli member of a novel subfamily of specialized Hsp70 chaperones Stefan Offermanns, Nina Wettschureck, Alexandra Zywietz G-Protein mediated signalling: Studies in conditional Galpha knockouts Andreas Plückthun Novel proteins from combinatorial and evolutionary approaches Matthias Gautschi, Sören Just, Sabine Rospert Chaperons at the Ribosomal Tunnel Exit - First Contacts of a Newly Synthesized Polypeptide R.T. Sauer, R. Burton, J. Kenniston, S. Joshi, D. Wah, I. Levchenko, R. Grant, J. Flynn, S. Neher, T. Baker ATP-dependent protein degradation and unfolding by ClpXP Andreas Strub, Karin Röttgers, Wolfgang Voos The peptide-binding domain determines the interaction of mitochondrial Hsp70s with the essential partner protein Tim44 Jörg Tatzelt, Konstanze F. Winklhofer The role of co- and posttranslational modifications in the propagation of scrapie-prions Ivo Zemp Circularly permuted proteins with alternative folding possibilities

Symposium: From protein transport to organelle development Ross Dalbey YidC, a novel evolutionarily conserved protein, mediates membrane protein assembly in bacteria Uener Kolukisaoglu, Burkhard Schulz, Bianca Hust, Ulf-Ingo Fluegge, Ralf Bernd Kloesgen, Michael Gutensohn Isolation of an Arabidopsis T-DNA mutant for the chloroplast protein import receptor atToc33 Johannes Herrmann, Tom Lutz, Stephan Meier, Marc Preuss, Gregor Szyrach, Frank Baumann Protein insertion into the inner membrane of mitochondria Ralf Bernd Klösgen, Bo Hou The mechanism of deltapH/TAT-dependent protein translocation across the thylakoid membrane Mario Jakob, Peter Hanner, Ralf Bernd Klösgen Hunting for proteins associated with the delta pH / Tat-pathway in chloroplasts Joao Pedro Marques, Ingrid Dudeck, Martin Schattat, Ralf Bernd Kloesgen Import and plastid sorting of chimaeric GFP-polypeptides Joao Pedro Marques, Ingrid Dudeck, Martin Schattat, Ralf Bernd Klösgen Targeting of chimaeric GFP-polypeptide fusions within chloroplasts Sabine Molik, Ralf Bernd Klösgen Transport and assembly of the Rieske Fe/S protein: What happens in the stromal space? Nils Wiedemann, Kaye Truscott, Peter Rehling, Chris Meisinger, Nikolaus Pfanner Mitochondrial translocases for precursor proteins Tom Rapoport Protein Transport In and Out of the ER Colin Robinson Structure and mechanism of the twin-arginine translocase Heinrich Jung, Maike Rochon, Corinna Weber-Sparenberg Targeting of proteins to the flagellar type III secretion system of Escherichia coli

Symposium: Mol. Mechanisms of development and differentiation of skeletal and cardiac muscles Thomas Braun Regulation of muscle cell development and regeneration by growth factors and bHLH proteins Manfred Frasch, Hsiu-Hsiang Lee, Patrick Lo, Ingolf Reim, Hideyuki Nagaso SIGNALING AND TRANSCRIPTIONAL CONTROL OF VISCERAL MUSCLE AND HEART DEVELOPMENT IN DROSOPHILA heiko funke-kaiser, julia lemmer, christian von langsdorff, alexander thomas, slobodan d. Kovacevic, meike strasdat, frank zollmann, martin paul, hans-dieter orzechowski The cardiac transcription factor Nkx2-5 regulates the alternative promoters of the endothelin-converting enzyme-1 gene in H9c2 cardiomyoblasts Mathias Gautel Novel signalling pathways in striated muscles Seigo Izumo The role of Csx/Nkx 2.5 homeobox gene in cardiac development and congenital heart disease. Michael Rudnicki, Patrick Seale, Atsushi Asakura, Anna Polesskaya, Anthony Scime Myogenic Specification of Adult Stem Cells in Skeletal Muscle Agapios Sachinidis, Jürgen Hescheler PDGF-BB induces selective differentiation of mouse embryonic stem cells to cardiac cells

Symposium: Mol. Structure, function and regulation of xenobiotic metabolising enzymes Michael Arand Structure-function relationships of epoxide hydrolases Matthias Bros, Ute Gödtel-Armbrust, Ulrich Förstermann, Jean-Paul Boissel CHARACTERIZATION OF THE MULTIPLE PROMOTERS OF THE HUMAN NEURONAL NITRIC OXIDE SYNTHASE GENE Jeroen Buters The knockout approach to understand the function of xenobiotic metabolising enzymes Karin Doods, Karl-Werner Scharmm, Antonius Kettrup INHIBITION OF EROD-ACTIVITY IN RAT HEPATOMA CELLS Kristin Ehrbar, Wolf-Dietrich Hardt Identification of the secretion and translocation domain of Salmonella typhimurium effector protein SopE Martin Göttlicher Nuclear receptors and their role in the regulation of xenobiotic metabolism Doris Kloor1, Angelika Lüdtke1, Stanka Stoeva2, Hartmut Osswald1 The ratio of the cofactor NAD +/NADH of S-adenosylhomocysteine hydrolase controls the adenosine binding sites. Paul Pfluger, Marc Birringer, Regina Brigelius-Flohé, Ralph Rühl, Nico Landes vitamin E

Symposium: Molecular medicine and functional genomics Eva Borst, Martin Messerle CYTOMEGALOVIRUS-A NEW VECTOR FOR GENE TRANSFER Lusine¹ Danielyan, Simone¹ Harsch, Christina¹ Schneider, Gayane² Buniatian, Heiner² Wiesinger, Selim³ Kuçi, Christoph¹ Gleiter Inverse Dynamics of Glial Fibrillary Acidic Protein (GFAP) Expression and Synthesis of Erythropoietin (EPO), EPO-Receptor (EPOR) and Proliferating Cell Nuclear Antigen (PCNA) in Astrocytes Sabine Groesch, Irmgard Tegeder, Karin Schilling, Ellen Niederberger, Gerd Geisslinger Activation of c-Jun-N-terminal-kinase by R- and S-flurbiprofen results in cell cycle arrest in human colon carcinoma cells Helmut Hanenberg, Detlev Schindler Fanconi anemia: basic molecular biology and clinical implications Regina Hühn, Beate Liebig, Martin S. Staege, Stefan Burdach Cleavage of EWS/FLI-1 fusion transcripts by Hammerhead Ribozymes Christoph Hutter, Silke Bergmann, Stefan Burdach, Martin S. Staege Analysis of EWS/Fli-1 induced gene expression in HEK293 cells using DNA-microarrays Christina Justenhoven, Hiltrud Brauch, Thomas Brüning, Hans-Peter Fischer, Ute Hamann, Volker Harth, Yon Ko, Beate Pesch Breast Cancer and Predictive Factors: Association with Polymorphisms Markus Kellner, Patrick Baer, Martin Wehling, Ralf Loesel Aldosterone action in humans -new members of the aldosterone signaling pathwaySabine Körber, Sandra Barth, Ulrich Schmidt, Christiane Jäger, Constanze Günther, Christoph Parthier, Gerald Böhm Polyomavirus-like particles as a system for gene therapy and drug delivery Tobias Kanacher, Miguel Pineda, Anita Schultz, Joachim Schultz, Jürgen Linder GAF-domains as novel cAMP-sensor autoactivate a bacterial adenylyl cyclase Maria Laura Massa, Simone Baltrusch, Sigurd Lenzen, Markus Tiedge Effect of PFK-2 overexpression in insulin-producing cells upon glucokinase activity and glucose metabolism Christian Monnerjahn, Ina Schüttmann, Manfred Konrad A designed variant of Herpes virus thymidine kinase selectively phosphorylates the anticancer and antiviral prodrug ganciclovir Stephan Ort, Manfred Konrad Structural and functional analysis of deoxyribonucleoside kinases dCK and dGK: design of novel mutant enzymes for therapeutic applications. Günther Richter, Karin Hanewinkel, Stefan Burdach Differential analysis of interleukin 2 (IL-2) versus IL-15 induced immediate-early genes: A contribution for the elucidation of the AICD paradoxon? Wiebke Scherf, Günther Richter, Jana Bergmann, Stefan Burdach, Gesine Hansen Nasal DNA vaccination with an Interleukin-10 cDNA vector in a murine asthma modell Hans J. Schramm Dimerization inhibitors of HIV-1 protease Stefan Welte, Günter Müller, Matthias Dreyer, Stefan Petry, Norbert Tennagels, Robert Schwenk Use of PTP1B mutants for selective inhibitor screening

Martin S. Staege, Christoph Hutter, Silke Bergmann, Sabine Körber, Stefan Burdach From genes to antigens: Potential target structures for autologous and allogeneic cytotoxic T cells identified by oligonucleotide arrays F. Westermann, J. S. Wei, M. Ringner, L. H. Saal, F. Berthold, M. Schwab, C Peterson, P Meltzer, J. Khan Classification and diagnostic prediction of pediatric cancers using gene expression profiling and artificial neural networks Stefan Wölfl, Annette Geyer, Larissa Odyvanova, Torsten Kroll, Peter Hortschansky, Manuela Schwalbe, Anke Naumann, Klaus Höffken, Joachim Clement Effect of BMP-2 on the migratory and invasive properties of breast cancer cells Albert Zlotnik, Anja Muller, Bernhard Homey A role for Chemokine Receptors in Cancer Metastasis?

Symposium: New Approaches in Cardiovascular Disease Andreas Busch NHE-1 inhibitors: From acute cardioprotection to heart failure Heyo K. Kroemer, Werner Siegmund, Ingolf Cascorbi Pharmacogenetics in the Cardiovascular Field Ulrich Laufs Significance of cholesterol-independent effects of statins Franz-Josef Neumann Molecular mechanisms of restenosis Meike Strasdat, Slobodan Kovacevic, Martin Paul, Anne Weinstrauch, Robert Real, Hans-Dieter Orzechowski Effect of MAP kinase inhibitors on injury-induced neointima formation in the rat carotid artery Andreas Simm, Sherif Daoud, Christian Casselmann, Rolf-Edgar Silber, Reinhard Schinzel Advanced Glycation Endproducts induced cell signalling in cardiac fibroblasts Peter Spangenberg, Nadine Zidek, Marion Grau, Isabel Göhring, Roy Eylenstein, Sven Güttich, Wolfgang Lösche, Stan Heptinstall Comparison of the effects of the P2Y12 ADP-receptor antagonist (AR-C69931) and the GPIIb/IIIa antagonist GR 144053F on platelet-leukocyte conjugate formation. Monika Stoll Genetic screening approaches for the identification of cardiovascular drug targets

Symposium: New aspects in GPCR-signalling Michel Bouvier Oligomeric assembly of G protein-coupled receptors: Role in receptor trafficking and function Otto-Erich Brodde, Rainer Büscher, Kirsten Leineweber, Heike Bruck Beta-adrenoceptor polymorphisms: does altered in vitro function predict in vivo effects? Arina Kostanian, Heinz Bönisch, Manfred Göthert, Michael Brüss Loss of function of the naturally occurring Arg219Leu variant of the human 5-HT1A receptor Marc Parmentier, Masato Kotani, Valérie Wittamer, David Communi, Isabelle Migeotte, Jalal Vakili, Michel Detheux, Stéphane Brézillon, Emmanuel Le Poul, Jean-Denis Franssen Natural ligand identification and functional characterization of orphan G protein-coupled receptors Detlef PIETROWSKI, Gia Anh Bao PHAN, Paulina WITT, Christoph KECK THE EXPRESSION OF ANGIOGENIC GROWTH FACTORS IN HUMAN GRANULOSA CELLS ARE CONTROLLED BY HUMAN CHORION GONADOTROPIN. Anton Vichalkovski, Karin Kirschner, Kurt Baltensperger, Hartmut Porzig SDF-1 and thrombin signaling in human hematopoietic progenitor cells is regulated by cytokines Rainer Schäfer, Georg Reiser Different regulation of human lung cancer cell proliferation by nucleotides mediated through P2Y receptors

Symposium: New molecular approaches to prevent infections Hannah Schröder-Borm, Regine Willumeit, Jörg Andrä Molecular basis for membrane selectivity of a potent peptide antibiotic derived from NK-lysin Leopold Flohé Molecular tools to design trypanocidal drugs Thomas Hartung G-CSF in infection prophylaxis Klaus Heeg Anti-infective intervention via Toll-like receptors Daniela Maennel Appropriate TNF production - a decisive factor in sepsis Hans-Dieter Volk, Susanna Prösch From the molecular mechanism of human cytomegalovirus (re)activation to new therapeutical concepts Susanna Prösch and Hans-Dieter Volk Institutes of Virology and Medical Immunology, Humboldt Unive Zuebeyde Pamukci, Andreas Koschinski, Eugen Domann, Trinad Chakraborty, Ayub Darji, Dierk Brockmeier, Florian Dreyer, Holger Repp, Jan Birringer Listeriolysin of Listeria monocytogenes forms Ca2+-permeable pores leading to intracellular Ca2+ oscillations in host cells

Symposium: Oxidative Stress and Redoxs-Regulation (SG Biochemische Pharmakologie u. Tox.) Kotb Abdelmohsen, Lars-Oliver Klotz, Helmut Sies Activation of EGF receptor-dependent signaling pathways by alkylating and redox-cycling quinones Kerstin Schnurr, Dagmar Kupper, Antje Banning, Cordula Müller, Matilde Maiorino, Gaby Böl, Regina Brigelius-Flohé Redox events in IL-1 signaling Yvonni Chovolou, Wim Wätjen, Andreas Kampkötter, Regine Kahl ALTERED SENSITIVITY OF TNF-ALPHA OVEREXPRESSING RAT HEPATOMA CELLS AGAINST EXOGENOUS TNF-ALPHA AND OXIDANTS Armin Kabat, Peter Rösen, Antje Olbrich, Stefan Dhein Effects of vitamin E on endothelial function in diabetes mellitus and hyperglycemia Harald F. Krug, Claudia Ball, Silvia Diabate Analysis of oxidative stress due to fly ash and ultrafine particles in macrophages and epithelial cells of the lung Bernd Epe Relevance of endogenous oxidative DNA damage: lessons from repair-deficient mice Peter Gierschik, Monika Gaugler, Ute Dreißigacker, Meike Müller, Klaudia Giehl K-Ras-dependent signal tranduction in human pancreatic carcinoma cells Arne Holmgren Redox regulation by thioredoxin and glutaredoxin systems Bernd Kammerer, Rainer Kahlich, Florence Pojer, Shuming Li, Christoph Gleiter, Lutz Heide Identification of aminocoumarin antibiotics from different Streptomyces species via ESI-MS and LC-MS/MS coupling Lars-Oliver Klotz Oxidant-induced signaling: from intra- to intercellular communication Carolin Oberle, Astrid Matzke, Silvia Meixner, Ulrich Massing, Harald F. Krug Alkylphosphocholine Analogues induce Apoptosis in Tumour Cells of the Immune System Wim Wätjen, Yvonni Chovolou, Andreas Kampkötter, Quynh-Hoa Tran-Thi, Regine Kahl, Petra Niering H2O2-INDUCED OXIDATIVE STRESS IN H4IIE CELLS: INFLUENCE OF THE FLAVONOID KAEMPFEROL Lars-Oliver Klotz, Helmut Sies, Peter Schröder (-)-Epicatechin Is Taken up by Cells and Protects against Peroxynitrite-Induced Oxidation and Nitration. Sandra Loeppen, Daniela Schneider, Frank Gaunitz, Rolf Gebhardt, Albrecht Buchmann, Michael Schwarz Promotion of hepatocarcinogenesis is mediated by overexpression of glutamine synthetase in beta-catenine-mutated mouse liver tumors Harald, F. Krug, Tanja Detzel, Siegfried Strack Involvement of different signalling pathways in cytoskeletal reorganisation during tributyltin (TBT) and H2O2 induced apoptosis Harald F. Krug, Silvia Diabaté, Katja Völkel Oxidative stress in rat alveolar epithelial cells and co-cultures upon treatment with fly ash Claudia von Montfort, Lars-Oliver Klotz Singlet Molecular Oxygen Reversibly Inhibits Tyrosine Phosphatases

Wim Wätjen, Yvonni Chovolou, Petra Niering, Andreas Kampkötter, Gudrun Aussel, Quynh-Hoa Tran-Thi, Regine Kahl PRO- AND ANTIAPOPTOTIC EFFECTS OF QUERCETIN AND FISETIN IN H4IIE-CELLS: IMPLICATION OF OXIDATIVE STRESS Margarita Fuhrmann, Kurt Racké, Folker Wenzel Reduced NO synthesis during halothane exposure in stimulated rat alveolar macrophages Josef H. Wissler, Enno Logemann Biomolecular switch from oxidative stress in inflammation to tissue morphogenesis in healing: Fenton-type OH* radical reactions and isoguanosine in metalloregulated RNA bioaptamers of S100-proteins Juergen Kuhlmann, Alexander Wolf Development of an in vitro system for detection and quantitation of urothelial genotoxicity of tobacco smoke-specific constituents

Symposium: Pharmacogenomics (Paul-Martini-Stiftung Symposium) Julian Arbuckle Pharmacogenetic based clinical trials to improve drug development Klaus-Peter Koller Genome wide search for disease genes as new drug targets Werner Kroll High throughput gene expression profiling to screen for drug effects and toxicity Andrè Reis Understanding the genetic basis of common human diseases and its potential for new concepts in drug therapy Dan Roden Implementing pharmacogenomics to improve drug therapy: where do we stand in 2002

Symposium: Pharmacogenomics (Paul-Martini-Stiftung Symposium) Julian Arbuckle Pharmacogenetic based clinical trials to improve drug development Klaus-Peter Koller Genome wide search for disease genes as new drug targets Werner Kroll High throughput gene expression profiling to screen for drug effects and toxicity Andrè Reis Understanding the genetic basis of common human diseases and its potential for new concepts in drug therapy Dan Roden Implementing pharmacogenomics to improve drug therapy: where do we stand in 2002

Symposium: Plenary Lectures Gunter Fischer Unfolding the Chemistry of Protein Folding Garret A. Fitzgerald The role of cyclooxygenases in the cardiovascular system Franz Oesch Toxicology from experimental systems to man: drug metabolising enzymes as key determinants Kai Simons Lipid rafts in membrane trafficking and cell polarity Kurt von Figura Inherited disorders caused by faulty protein modification in the secretory route

Symposium: Oxidative Stress and Redoxs-Regulation (SG Biochemische Pharmakologie u. Tox.) Kotb Abdelmohsen, Lars-Oliver Klotz, Helmut Sies Activation of EGF receptor-dependent signaling pathways by alkylating and redox-cycling quinones Kerstin Schnurr, Dagmar Kupper, Antje Banning, Cordula Müller, Matilde Maiorino, Gaby Böl, Regina Brigelius-Flohé Redox events in IL-1 signaling Yvonni Chovolou, Wim Wätjen, Andreas Kampkötter, Regine Kahl ALTERED SENSITIVITY OF TNF-ALPHA OVEREXPRESSING RAT HEPATOMA CELLS AGAINST EXOGENOUS TNF-ALPHA AND OXIDANTS Armin Kabat, Peter Rösen, Antje Olbrich, Stefan Dhein Effects of vitamin E on endothelial function in diabetes mellitus and hyperglycemia Harald F. Krug, Claudia Ball, Silvia Diabate Analysis of oxidative stress due to fly ash and ultrafine particles in macrophages and epithelial cells of the lung Bernd Epe Relevance of endogenous oxidative DNA damage: lessons from repair-deficient mice Peter Gierschik, Monika Gaugler, Ute Dreißigacker, Meike Müller, Klaudia Giehl K-Ras-dependent signal tranduction in human pancreatic carcinoma cells Arne Holmgren Redox regulation by thioredoxin and glutaredoxin systems Bernd Kammerer, Rainer Kahlich, Florence Pojer, Shuming Li, Christoph Gleiter, Lutz Heide Identification of aminocoumarin antibiotics from different Streptomyces species via ESI-MS and LC-MS/MS coupling Lars-Oliver Klotz Oxidant-induced signaling: from intra- to intercellular communication Carolin Oberle, Astrid Matzke, Silvia Meixner, Ulrich Massing, Harald F. Krug Alkylphosphocholine Analogues induce Apoptosis in Tumour Cells of the Immune System Wim Wätjen, Yvonni Chovolou, Andreas Kampkötter, Quynh-Hoa Tran-Thi, Regine Kahl, Petra Niering H2O2-INDUCED OXIDATIVE STRESS IN H4IIE CELLS: INFLUENCE OF THE FLAVONOID KAEMPFEROL Lars-Oliver Klotz, Helmut Sies, Peter Schröder (-)-Epicatechin Is Taken up by Cells and Protects against Peroxynitrite-Induced Oxidation and Nitration. Sandra Loeppen, Daniela Schneider, Frank Gaunitz, Rolf Gebhardt, Albrecht Buchmann, Michael Schwarz Promotion of hepatocarcinogenesis is mediated by overexpression of glutamine synthetase in beta-catenine-mutated mouse liver tumors Harald, F. Krug, Tanja Detzel, Siegfried Strack Involvement of different signalling pathways in cytoskeletal reorganisation during tributyltin (TBT) and H2O2 induced apoptosis Harald F. Krug, Silvia Diabaté, Katja Völkel Oxidative stress in rat alveolar epithelial cells and co-cultures upon treatment with fly ash Claudia von Montfort, Lars-Oliver Klotz Singlet Molecular Oxygen Reversibly Inhibits Tyrosine Phosphatases

Wim Wätjen, Yvonni Chovolou, Petra Niering, Andreas Kampkötter, Gudrun Aussel, Quynh-Hoa Tran-Thi, Regine Kahl PRO- AND ANTIAPOPTOTIC EFFECTS OF QUERCETIN AND FISETIN IN H4IIE-CELLS: IMPLICATION OF OXIDATIVE STRESS Margarita Fuhrmann, Kurt Racké, Folker Wenzel Reduced NO synthesis during halothane exposure in stimulated rat alveolar macrophages Josef H. Wissler, Enno Logemann Biomolecular switch from oxidative stress in inflammation to tissue morphogenesis in healing: Fenton-type OH* radical reactions and isoguanosine in metalloregulated RNA bioaptamers of S100-proteins Juergen Kuhlmann, Alexander Wolf Development of an in vitro system for detection and quantitation of urothelial genotoxicity of tobacco smoke-specific constituents

Symposium: Pharmacogenomics (Paul-Martini-Stiftung Symposium) Julian Arbuckle Pharmacogenetic based clinical trials to improve drug development Klaus-Peter Koller Genome wide search for disease genes as new drug targets Werner Kroll High throughput gene expression profiling to screen for drug effects and toxicity Andrè Reis Understanding the genetic basis of common human diseases and its potential for new concepts in drug therapy Dan Roden Implementing pharmacogenomics to improve drug therapy: where do we stand in 2002

Symposium: Pharmacogenomics (Paul-Martini-Stiftung Symposium) Julian Arbuckle Pharmacogenetic based clinical trials to improve drug development Klaus-Peter Koller Genome wide search for disease genes as new drug targets Werner Kroll High throughput gene expression profiling to screen for drug effects and toxicity Andrè Reis Understanding the genetic basis of common human diseases and its potential for new concepts in drug therapy Dan Roden Implementing pharmacogenomics to improve drug therapy: where do we stand in 2002

Symposium: Plenary Lectures Gunter Fischer Unfolding the Chemistry of Protein Folding Garret A. Fitzgerald The role of cyclooxygenases in the cardiovascular system Franz Oesch Toxicology from experimental systems to man: drug metabolising enzymes as key determinants Kai Simons Lipid rafts in membrane trafficking and cell polarity Kurt von Figura Inherited disorders caused by faulty protein modification in the secretory route

Symposium: Proteins involved in apoptosis and transport Jörg Kahle, Marc Bäuerle, Matthias Baake, Detlef Doenecke, Werner Albig Nuclear import of histones Ralf Baumann, Hans-Uwe Simon MACROPHAGE MIGRATION INHIBITORY FACTOR (MIF) IS A SURVIVAL FACTOR FOR NEUTROPHILS Simone Fulda, Klaus-Michael Debatin Therapeutic modulation of death receptor- or drug-mediated apoptosis Klaus Heese, Yasuo Nagai, Tohru Sawada Characterizing the signaling pathway of rat p18 amyloid beta (Aß) responsive protein (p18AßrP). Matthias Kappler, Matthias Kotzsch, Frank Bartel, Susanne Füssel, Chritine Lautenschläger, Peter Würl, Matthias Bache, Hannelore Schmidt, Helge Taubert, Axel Meye Elevated expression level of survivin protein in soft-tissue sarcomas Dietrich Keppler, Yunhai Cui, Jörg König, Anne T. Nies Proteins mediating vectorial transport across plasma membrane domains Harald F. Krug, Silvia Meixner Caspase-10 is the Key Initiator-Caspase involved in TBT-mediated Apoptosis Anne Navarrete Santos, Sarah Tonack, Michaela Kirstein, Bernd Fischer The insulin receptor and the insulin-dependent glucose transporter isoform 4 are expressed in rabbit preimplantation embryos Ludwig Nißler Binding of flavonoid and anthraquinone derivatives to nucleotide binding domains of ABC transporters. Ernst Petzinger, Kerstin Lischka, Dieter Starke Secretion of intact oligonucleotides by mrp2 into bile Volker Florian, Peter Knauth, Thomas Schlüter, Susan Schreckenberger Characterisation of Protein Domains Responsible for Recognition of P-Selectin by Sorting Nexin 17 Kent Söe, Hella Hartmann, Holger Stephan, Bernhard Schlott, Tinna Stevnsner*, Frank Grosse p53 is involved in the repair of human topoisomerase I cleavage complexes Ekkehardt Stehfest, Abdel Torky, Heidi Foth MRP expression and modulation of/by glutathione in human lung cells christopher stroh, ajoy samraj, uwe cassens, klaus schulze-osthoff, marek los Caspase inhibition strongly improves the recovery of cryopreserved hematopoietic and other cells

Symposium: Proteins involved in apoptosis and transport Jörg Kahle, Marc Bäuerle, Matthias Baake, Detlef Doenecke, Werner Albig Nuclear import of histones Ralf Baumann, Hans-Uwe Simon MACROPHAGE MIGRATION INHIBITORY FACTOR (MIF) IS A SURVIVAL FACTOR FOR NEUTROPHILS Simone Fulda, Klaus-Michael Debatin Therapeutic modulation of death receptor- or drug-mediated apoptosis Klaus Heese, Yasuo Nagai, Tohru Sawada Characterizing the signaling pathway of rat p18 amyloid beta (Aß) responsive protein (p18AßrP). Matthias Kappler, Matthias Kotzsch, Frank Bartel, Susanne Füssel, Chritine Lautenschläger, Peter Würl, Matthias Bache, Hannelore Schmidt, Helge Taubert, Axel Meye Elevated expression level of survivin protein in soft-tissue sarcomas Dietrich Keppler, Yunhai Cui, Jörg König, Anne T. Nies Proteins mediating vectorial transport across plasma membrane domains Harald F. Krug, Silvia Meixner Caspase-10 is the Key Initiator-Caspase involved in TBT-mediated Apoptosis Anne Navarrete Santos, Sarah Tonack, Michaela Kirstein, Bernd Fischer The insulin receptor and the insulin-dependent glucose transporter isoform 4 are expressed in rabbit preimplantation embryos Ludwig Nißler Binding of flavonoid and anthraquinone derivatives to nucleotide binding domains of ABC transporters. Ernst Petzinger, Kerstin Lischka, Dieter Starke Secretion of intact oligonucleotides by mrp2 into bile Volker Florian, Peter Knauth, Thomas Schlüter, Susan Schreckenberger Characterisation of Protein Domains Responsible for Recognition of P-Selectin by Sorting Nexin 17 Kent Söe, Hella Hartmann, Holger Stephan, Bernhard Schlott, Tinna Stevnsner*, Frank Grosse p53 is involved in the repair of human topoisomerase I cleavage complexes Ekkehardt Stehfest, Abdel Torky, Heidi Foth MRP expression and modulation of/by glutathione in human lung cells christopher stroh, ajoy samraj, uwe cassens, klaus schulze-osthoff, marek los Caspase inhibition strongly improves the recovery of cryopreserved hematopoietic and other cells

Symposium: Proteins involved in apoptosis and transport Jörg Kahle, Marc Bäuerle, Matthias Baake, Detlef Doenecke, Werner Albig Nuclear import of histones Ralf Baumann, Hans-Uwe Simon MACROPHAGE MIGRATION INHIBITORY FACTOR (MIF) IS A SURVIVAL FACTOR FOR NEUTROPHILS Simone Fulda, Klaus-Michael Debatin Therapeutic modulation of death receptor- or drug-mediated apoptosis Klaus Heese, Yasuo Nagai, Tohru Sawada Characterizing the signaling pathway of rat p18 amyloid beta (Aß) responsive protein (p18AßrP). Matthias Kappler, Matthias Kotzsch, Frank Bartel, Susanne Füssel, Chritine Lautenschläger, Peter Würl, Matthias Bache, Hannelore Schmidt, Helge Taubert, Axel Meye Elevated expression level of survivin protein in soft-tissue sarcomas Dietrich Keppler, Yunhai Cui, Jörg König, Anne T. Nies Proteins mediating vectorial transport across plasma membrane domains Harald F. Krug, Silvia Meixner Caspase-10 is the Key Initiator-Caspase involved in TBT-mediated Apoptosis Anne Navarrete Santos, Sarah Tonack, Michaela Kirstein, Bernd Fischer The insulin receptor and the insulin-dependent glucose transporter isoform 4 are expressed in rabbit preimplantation embryos Ludwig Nißler Binding of flavonoid and anthraquinone derivatives to nucleotide binding domains of ABC transporters. Ernst Petzinger, Kerstin Lischka, Dieter Starke Secretion of intact oligonucleotides by mrp2 into bile Volker Florian, Peter Knauth, Thomas Schlüter, Susan Schreckenberger Characterisation of Protein Domains Responsible for Recognition of P-Selectin by Sorting Nexin 17 Kent Söe, Hella Hartmann, Holger Stephan, Bernhard Schlott, Tinna Stevnsner*, Frank Grosse p53 is involved in the repair of human topoisomerase I cleavage complexes Ekkehardt Stehfest, Abdel Torky, Heidi Foth MRP expression and modulation of/by glutathione in human lung cells christopher stroh, ajoy samraj, uwe cassens, klaus schulze-osthoff, marek los Caspase inhibition strongly improves the recovery of cryopreserved hematopoietic and other cells

Symposium: Proteomics (SG Mol. Zellbiologie, SG Bioanalytik) Frank Bier On-chip synthesis of extended, long-chain DNA by PCR Xenia Marschan, Dennie Andresen, Markus von Nickisch-Rosenegk, Frank F. Bier Ljudmila V. Borissenko, Qinghua Fang, Jens Fey, Thomas Dierks, Kurt von Figura C(alpha)-Formylglycine: a novel protein modification in pro- and eukaryotes Cordula Enenkel Targeting mechanism of proteasomes to nuclear / ER membranes and into the nucleus of yeast Anne-Claude Gavin, Paola Grandi, Markus Boesche, Roland Krause, Christina Leutwein, Bernhard Kuster, Giulio Superti-Furga Functional organization of the yeast proteome by systemic analysis of protein complexes Wolfgang Hilt, Iris Velten, Martin Ligr, Harish Karnam, Julia Ilyina Functions of the proteasome in cell cycle and apoptosis Reiner Lammers, Hans-Ulrich Häring, Katja Kapp Cellular transformation mediated by Src kinase: the role of PTPalpha Steffen Liedtke The Use of Multidimensional Capillary LC in Proteomics Heike Schaefer, Yvonne Wagner, Albert Sickmann, Helmut E. Meyer* High throughput mass spectrometry for proteomics Daniel de Graaf, Philip Denner, Luiza Bengtsson, Henning Otto Characterization of Inner Nuclear Membrane Proteins Peter Rehling Molecular Dissection of the Protein Import Complexes of the Inner Membrane of Yeast Mitochondria Ulrike D. Epple, Henning Barth, Michael Thumm Aut10p, Mai1p and Aut5p shed new lights on the molecular mechanisms of autophagy in S.cerevisiae

Symposium: Transfection with proteins and antisense-RNA Frank Czubayko, Achim Aigner Ribozyme-PEI-complexes as ready-to-use reagents for efficient down-regulation of gene expression Navarrete Santos Alexander, Immisch Claudia, Reinke Claudia, Schaupp Michael, Böhm Gerald On the use of a histone-like protein for drug delivery using doxorubicin as a model system for tumor therapy Frank Bartel, Peter Würl, Axel Meye, Matthias Kappler, Matthias Bache, Hannelore Schmidt, Manfred Schönfelder, Helge Taubert Growth reduction of a xenotransplanted human Soft Tissue Sarcoma by MDM2 Antisense Therapy Lars Boenicke, Regina Pauls, Chu Kang, Holger Kalthoff Efficient Dosage and Time dependent Protein Transduction of Pancreatic Carcinoma Cells using purified VP22-EGFP Fusion Protein M. Cristina Cardoso, Wolfgang Derer, Hariharan P. Easwaran, Heinrich Leonhardt Protein therapy: Applications in tissue regeneration and stem cell research Karsten Hemmrich, Christoph V. Suschek, Guido Lerzynski, Victoria Kolb-Bachofen OPTIMIZED ANTISENSE OLIGONUCLEOTIDE-MEDIATED INHIBITION OF THE INDUCIBLE NITRIC OXIDE SYNTHASE CO-SUPPRESSES NO-MEDIATED STRESS RESPONSE IN ENDOTHELIUM Claudia Immisch, Alexander Navarrete Santos, Gerald Böhm Binding of RNA to the hyperthermophilic histone-like protein TmHU Michael Metzlaff Transgene- and virus-mediated gene silencing - What plants can teach us! XiaoLi Lu, Zhibao Mi, Jeff Mai, Paul Robbins Characterization, Optimization and Application of Peptide-Mediated Protein Transduction Thomas Tuschl, Sayda M. Elbashir, Jens Harborth, Kim Bechert, Mariana Lagos-Quintana, Javier Martinez, Abdullah Yalcin, Agnieszka Patkaniowska, Jutta Meyer, Klaus Weber A new world of tiny RNAs - siRNAs and miRNAs

Symposium: Proteomics (SG Mol. Zellbiologie, SG Bioanalytik) Frank Bier On-chip synthesis of extended, long-chain DNA by PCR Xenia Marschan, Dennie Andresen, Markus von Nickisch-Rosenegk, Frank F. Bier Ljudmila V. Borissenko, Qinghua Fang, Jens Fey, Thomas Dierks, Kurt von Figura C(alpha)-Formylglycine: a novel protein modification in pro- and eukaryotes Cordula Enenkel Targeting mechanism of proteasomes to nuclear / ER membranes and into the nucleus of yeast Anne-Claude Gavin, Paola Grandi, Markus Boesche, Roland Krause, Christina Leutwein, Bernhard Kuster, Giulio Superti-Furga Functional organization of the yeast proteome by systemic analysis of protein complexes Wolfgang Hilt, Iris Velten, Martin Ligr, Harish Karnam, Julia Ilyina Functions of the proteasome in cell cycle and apoptosis Reiner Lammers, Hans-Ulrich Häring, Katja Kapp Cellular transformation mediated by Src kinase: the role of PTPalpha Steffen Liedtke The Use of Multidimensional Capillary LC in Proteomics Heike Schaefer, Yvonne Wagner, Albert Sickmann, Helmut E. Meyer* High throughput mass spectrometry for proteomics Daniel de Graaf, Philip Denner, Luiza Bengtsson, Henning Otto Characterization of Inner Nuclear Membrane Proteins Peter Rehling Molecular Dissection of the Protein Import Complexes of the Inner Membrane of Yeast Mitochondria Ulrike D. Epple, Henning Barth, Michael Thumm Aut10p, Mai1p and Aut5p shed new lights on the molecular mechanisms of autophagy in S.cerevisiae

Symposium: Transfection with proteins and antisense-RNA Frank Czubayko, Achim Aigner Ribozyme-PEI-complexes as ready-to-use reagents for efficient down-regulation of gene expression Navarrete Santos Alexander, Immisch Claudia, Reinke Claudia, Schaupp Michael, Böhm Gerald On the use of a histone-like protein for drug delivery using doxorubicin as a model system for tumor therapy Frank Bartel, Peter Würl, Axel Meye, Matthias Kappler, Matthias Bache, Hannelore Schmidt, Manfred Schönfelder, Helge Taubert Growth reduction of a xenotransplanted human Soft Tissue Sarcoma by MDM2 Antisense Therapy Lars Boenicke, Regina Pauls, Chu Kang, Holger Kalthoff Efficient Dosage and Time dependent Protein Transduction of Pancreatic Carcinoma Cells using purified VP22-EGFP Fusion Protein M. Cristina Cardoso, Wolfgang Derer, Hariharan P. Easwaran, Heinrich Leonhardt Protein therapy: Applications in tissue regeneration and stem cell research Karsten Hemmrich, Christoph V. Suschek, Guido Lerzynski, Victoria Kolb-Bachofen OPTIMIZED ANTISENSE OLIGONUCLEOTIDE-MEDIATED INHIBITION OF THE INDUCIBLE NITRIC OXIDE SYNTHASE CO-SUPPRESSES NO-MEDIATED STRESS RESPONSE IN ENDOTHELIUM Claudia Immisch, Alexander Navarrete Santos, Gerald Böhm Binding of RNA to the hyperthermophilic histone-like protein TmHU Michael Metzlaff Transgene- and virus-mediated gene silencing - What plants can teach us! XiaoLi Lu, Zhibao Mi, Jeff Mai, Paul Robbins Characterization, Optimization and Application of Peptide-Mediated Protein Transduction Thomas Tuschl, Sayda M. Elbashir, Jens Harborth, Kim Bechert, Mariana Lagos-Quintana, Javier Martinez, Abdullah Yalcin, Agnieszka Patkaniowska, Jutta Meyer, Klaus Weber A new world of tiny RNAs - siRNAs and miRNAs

Symposium: _Other (free) topic_ Jürgen Alves, Dorothee von Witzendorff, Minh-Cam Ha-Thi, Susanne von Pall de Tolna, Gabriele Grabowski Restriction endonuclease EcoRI - fusions for a change of specificity Stefanie Barbirz The tailspike from E.Coli H bacteriophage HK620: Another Parallel beta-Helix Protein? Mike Beck, Kieran Brickley, Miriam Smith, Seema Sharma, Helen Wilkinson, F. Anne Stephenson GRIF-1 - a novel GABAA receptor associated protein Daniela Besong Genexpression of the human membrane-associated Progesterone Receptor (hmPR) in HepG2 cells Susanne Brakmann, Sylvia Löbermann, Petra Nieckchen Optimal enzymes for single molecule sequencing Peter Rehling, Peter Kovermann, Richard Wagner, Nikolaus Pfanner, Kaye, N. Truscott, Katrin Brandner The TIM22 translocase mediates insertion of carrier precursors into the mitochondrial inner membrane via a two-step process S. Breidert, R. Jacob, A. Ngezahajo, H.-A. Kolb, H. Naim Trafficking pathways of connexin49-GFP in living mammalian cells Claudia Loske, Gerald Münch, Björn Kuhla, Sladjana Dukic-Stefanovic Differential effects of "advanced glycation endproducts" and ß-amyloid peptide on glucose utilization and ATP levels in the neuronal cell line SH-SY5Y Sladjana Dukic-Stefanovic, Jovana Gasic-Milenkovic, Winnie Deuther-Conrad, Gerald Münch Proinflammatory signal transduction pathways in N11 mouse microglial cell line activated by "Advanced Glycation Endproducts" Johanna Mansfeld, Renate Ulbrich-Hofmann, Peter Dürrschmidt Identification of an intermediate in the unfolding of neutral protease from Bacillus stearothermophilus by fluorescence spectroscopy Matthias Eckhardt, Simon Fewou, Volkmar Gieselmann Effect of N-glycan removal on catalytic activity of cerebroside sulfotransferase Jörg Bär, Jürgen Kirchberger, Anke Edelmann Interactions between the two types of subunits forming an enzymatically active oligomeric phosphofructokinase-1 from Saccharomyces cerevisiae Ralph-Peter Elsner Solution structure of the Ras binding domain of AF6 Jochen Frenzel, Jan Richter, Klaus Eschrich Nitrogen oxide-induced cell death in rabbit cultured Müller cells is modulated by glucose Leopold Flohé The element of the moon moonlighting for fertility Bernd Moritz, Gudrun Franke, Martin Siemann, Matthias Reuss, Helmut E. Meyer Quantitative Proteomics with Escherichia coli Alexander Freiberg, Jan Lengefeld, Matthias Machner, Wolfgang Pfeil, Dirk Heinz, Robert Seckler Stability and Folding of Internalin B - A Biophysical Characterization Susana Gariba de Garcia, Gerald Münch, Gabriele Oehme

"Carbonyl stress” by methylglyoxal causes mitochondrial dysfunction, ATP depletion and NMDA receptor overstimulation in neurons Jovana Gasic-Milenkovic, Sladjana Dukic-Stefanovic, Winnie Deuther-Conrad, Gerald Münch ß-amyloid peptide potentiates inflammatory responses induced by lipopolysaccharide, interferon-gamma and "advanced glycation endproducts" in a mouse microglia cell line Klaus Golka, Seidel Thilo, Roetzel Claudia, Geller Frank, Bolt Hermann M, Dietrich Holger, Foth Heidi, Thier Ricarda Enzym polymorphisms and risk factors in bladder cancer patients in Lutherstadt Wittenberg Oliver Ohlenschläger, Jens Wöhnert, Enrico Bucci, Ramadurai Ramachandran, Roland Zell, Matthias Görlach Structure of an RNA Involved in Enteroviral Replication Anke Hannemann, Marina Bigl, Frank Gaunitz, Klaus Eschrich Characterization of the human platelet-type 6-phosphofructo-1-kinase gene promoter Monika Walter, Robert Seckler, Benjamin Heinz Folding Kinetics of Pectate Lyase from Bacillus subtilis Susanne Ammon, Peter Mayer, Volker Höllt Microarray analysis fo genes expressed in the frontal cortex of rats chronically treated with morphine Gudrun Horn, Daniel Weinfurtner, Roland Hofweber, Robert Macek, Robert Gail, Till Maurer, Hans Robert Kalbitzer Optimization of intein based isotope labeling methods for NMR investigations of proteins Franziska Bleichert*, J.P. Warnke**, Klaus Eschrich*, Renate Keßler* Expression of ubiquitous 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase in intracranial neoplasms and non-tumorous brain tissue V. Krügel Inhibition of LiCl-induced induction of glutamine synthetase in HepG2 cells by flavonoids and anthraquinone derivatives is mediated by CK I Peter Kühl Who discovered the Michaelis-Menten hyperbola? Antonios Kyriakopoulos, Barbara Hoppe, Alexandra Graebert, Marcus Kühbacher, Dietrich Behne Selenoproteins in the perinuclear structures of rat kidney Dedt. Molecular Trace Element research in the Life Sciences Hahn- Meitner-Istitut, Glienicker Str.100, 14109 Berlin Carsten Skarke, Helmut Schmidt, Jürgen Liefhold, Gerd Geisslinger, Jörn Lötsch A mathematical model to predict plasma concentrations of morphine and its active metabolite morphine-6-glucuronide in healthy young persons Johanna Mansfeld, Eva Petermann, Renate Ulbrich-Hofmann Generation of catalytically active neutral protease from Bacillus stearothermophilus does not require the presence of the propeptide Walter-Vesely Seb. Meister Polyvinylnucleobases: Synthesis, characterization and applications in nanotechnology Nils Hanekop, Susanne Wied, Stefan Petry, Norbert Tennagels, Wendelin Frick, Günter Müller A Receptor for Glycolipid Headgroups Mediates Insulin-mimetic Signaling in Rat Adipocytes Pavel I. Nedvetsky, Alexander Y. Kots, Helmut Müller, Ferid Murad, Harald H.H.W. Schmidt Hsp90 effects on the regulation of soluble guanylyl cyclase Ellen Niederberger, Achim Schmidtko, Jeffrey D. Rothstein, Gerd Geisslinger, Irmgard Tegeder Modulation of spinal nociceptive processing through the glutamate transporter GLT-1

Kathrin Fuchs, Diana Kühn, Johannes Schmitt, Joachim Nöller Membrane affinity and pore forming properties of proteins/peptides - Easy and fast by using solid supported lipid bilayers. Ingo Paarmann, Manfred Konrad Structural requirements for calmodulin binding to membrane-associated guaylate kinases Thomas Schneider, Gerhard Braus, Gabriele Heinrich, Markus Hartmann, Andrea Pfeil Evolution of Feedback-inhibited (&beta/&alpha)8 Barrel Isoenzymes by Gene Duplication and A Single Mutation Gia Anh Bao PHAN, Detlef PIETROWSKI, Christoph KECK EXPRESSION OF A NEW SPLICE VARIANT OF IGFBP-7/MAC25 TUMOR SUPPRESSOR GENE IN HUMAN GRANULOSA CELLS LACKING THE IGFBP MOTIF (GCGCCXXC) Paola Pocar, Robert Augustin, Bernd Fischer Induction of apoptotic cell death in bovine cumulus-oocyte complexes after treatment with Aroclor-1254 during in vitro maturation. Albert Sickmann, Stefan Lehr, Armin Herkner, Dirk Müller-Wieland, Helmut E. Meyer, Jörg Reinders Identification of the Major Tyrosin Phosphorylation Sites of Human Gab-1 Jens Wulfaenger, Alexander Navarrete Santos, Tanja Blosz, Juergen Langner, Dagmar Riemann The intracellular N-terminus of aminopeptidase N/CD13 affects association with caveolae Ina Wittko, Sigrid Saaler-Reinhardt Involvement of La protein in alternative translation processes in neural progenitor cells Polin Mahjour, Katja Mühlberg, Andreas Hagendorff, Stefan Dhein, Dietrich Pfeiffer, Aida Salameh Differential regulation of gap junction protein expression in cardiomyocytes Albert Sickmann, Marion Herrmann, Joachim Klose, Helmut E. Meyer, Heike Schaefer Analysis of Posttranslational Modifications of alpha-A-Crystallin during Aging of the Eye Lens Martin Haßler, Edgar Brändle, Joachim Greven, Jan Henrik Schlattjan Interaction of the organic base cimetidine with the renal basolateral p-aminohippurate (PAH) transport system Heide Schmid, Thai-Hoa Nguyen-Xuan, Minh-Huy Tran, Gerold Schwarz, Hubert Kalbacher, Heinrich Wiesinger The endosomal/lysosomal distribution of cathepsins (cat) B, L and S and N-acetyl-ß-D-glucosaminidase (NAG) is differentially modulated in murine cell lines of microglial and monocytic origin Marcel Schmitt, Georg Gellert, Bettina Kirberg, Jost Ludwig, Hella Lichtenberg-Fraté CyGene: Eine neue hefebasierte Methode zur Bestimmung des cyto- und genotoxischen Potentials von Umweltgiften im Wasserbereich. CyGene: A New Yeast Based Method for the Detection of Cyto- and Genotox Reinhild Tenderich, Brigitte Schmitz Function of the PEST sequence of NCAM 140 Astrid Schön, Olaf Gimple, Christian Heubeck The evolution of RNase P in eukaryotic organelles Lorenz Trümper, Frank Griesinger, Christa Fonatsch, Astrid Heutelbeck, Detlef Haase, Ernst Hallier, Thomas Schulz GLUTATHIONE S-TRANSFERASE GSTP1 POLYMORPHISMS IN DE NOVO AND THERAPY-INDUCED HEMATOLOGIC MALIGNANCIES Gabi Bauer-Manz, Andreas Kuhn, Lambertus van den Berg, Justyna Serek The E.coli membrane insertase YidC: A Chaperone for Protein Folding into the Membrane

Michael Spoerner, Guido Steiner, Thomas Prisner, Alfred Wittinghofer, Hans-Robert Kalbitzer Are GTP-analogs really good analogs? Anna Stepczynska, Jakob Troppmair, Thomas Voegel, Monika Poppe, Klaus Schulze-Osthoff, Michael Schwarz Bcl-2 antagonist of cell death (Bad) retains its proapoptotic activity despite proteolytic cleavage by caspase-3 Stefan Henrich, Robert Huber, Albert Ries, Karlheinz Mann, Klaus Kühn, Rupert Timpl, Gleb P. Bourenkov, Hans D. Bartunik, Wolfram Bode, Manuel E. Than Crystal structure of the noncollagenous (NC1) domain of human placenta collagen IV: Stabilization via a novel type of covalent Met-Lys cross-link Cornelia Körting, Alexander Froschauer, Reinhold Hanel, Jean-Nicolas Volff, Anne-Marie Veith DMRT Genes and Sex Determination in the Fish Xiphophorous Hubert Zipper, Katrin Lämmle, Christiane Buta, Herwig Brunner, Jürgen Bernhagen, Frank Vitzthum Investigations on the binding of SYBR Green I to double-stranded (ds)DNA Markus von Nickisch-Rosenegk, Jenny Steffen, Frank F. Bier Transcription of Reporter Genes with Immobilized Templates Melanie Wagner, Reinhard Reents, David Owen, Herbert Waldmann, Alfred Wittinghofer, Jürgen Kuhlmann Localisation of fluorescently labelled Ras protein in the living cell Albert Sickmann, Helmut E. Meyer, Yvonne Wagner Two-dimensional chromatography in protein analysis Robert Seckler, Monika Walter Pectate Lyase from Bacillus subtilis (BsPel): Mutations in the asparagine ladder cause a temperature-sensitive folding phenotype Monika Walter Pectate Lyase from Bacillus subtilis (BsPel): Mutations in the asparagine ladder cause a temperature-sensitive folding phenotype Sabine Werner, Stefan Kuklinski, Brigitte Schmitz Altered O-GlcNAc level after neuronal differentiation of PC12 cells Barth Holger, Klaus Aktories, Christian Wilde Clostridium botulinum C2 toxin as a protein transport system Klaus Aktories, Christian Wilde Clostridium botulinum C3 exoezyme like ADP-ribosyltransferases Jens Wulfänger, Tanja Blosz, Alexander Navarrete Santos, Jürgen Langner, Dagmar Riemann Aminopeptidase N/CD13 is associated with Lubrol rafts Ulrike Sattler, Ingrid Koziollek-Drechsler, Danuta Dormann, Christina Zechel Putative role of the nuclear factor NCNF in neuronal differentiation Suisheng Zhang, Carsten Köhler, Frank Grosse PHYSICAL INTERACTION BETWEEN NUCLEAR DNA HELICASE II AND WRN HELICASE STIMULATES WRN´S EXONUCLEASE ACTIVITY

Susana Gariba de Garcia, Gerald Münch, Gabriele Oehme "Carbonyl stress” by methylglyoxal causes mitochondrial dysfunction, ATP depletion and NMDA receptor overstimulation in neurons Advanced glycation endproducts (AGEs), di(carbonyl)-derived compounds, are found in "neurofibrillary tangles", characteristic intraneuronal protein deposits of Alzheimer‘s disease (AD), suggesting that crosslinking-reactive dicarbonyl compounds such as methylglyoxal (MG) are elevated in affected neurons [1]. MG is a very reactive dicarbonyl compound, and is formed by non-enzymatic degradation of triosephosphates. It is accumulated under conditions of decreased metabolic utilisation of triosephosphates, or by impaired MG detoxification occurring under conditions of GSH depletion [2]. The aim of this study was to investigate cellular effects of MG using a model neuronal cell line (SH-SY5Y neuroblastoma cells, which are, like many cancer cells particularly sensitive to MG toxicity). Furthermore, protective effects of carbonyl scavengers, antioxidants, nitric oxide synthase inhibitors and NMDA receptor antagonists were tested. MG (applied in concentrations ranging from 70 - 2500 µM) caused cell death, a decrease in intracellular ATP content, mitochondrial membrane depolarization and increased formation of mitochondrial reactive oxygen and nitrogen species in a concentration-dependent manner. MG induced reduction in cell number was accompanied by a decrease in the number and length of neurites. ATP depletion could be significantly prevented by the carbonyl scavengers aminoguanidine, tenilsetam, carnosine; and by the NMDA receptor antagonists MK-801, Memantine, and D-AP7. Thus, scavenging of dicarbonyl compounds or interference with their downstream pathways may provide new therapeutic opportunities to reduce the pathophysiological changes associated with dicarbonyl derived neurodegeneration. Literature [1] Wong A, Lüth HJ, Arendt Th, Münch G (2001) Advanced glycation endproducts co-localize with inducible nitric oxide synthase in Alzheimer’s disease. Brain Res 920: 32-40. [2] Thornalley PJ, Jahan I, Ng R. (2001) Suppression of the accumulation of triosephosphates and increased formation of methylglyoxal in human red blood cells during hyperglycaemia by thiamine in vitro. J Biochem (Tokyo) 129: 543-9. contact: PD Dr. Gerald Münch IZKF Leipzig Neuroimmunologische Zellbiologie [email protected] Johannisallee 30a 04103 Leipzig (Germany)

Kathrin Fuchs, Diana Kühn, Johannes Schmitt, Joachim Nöller Membrane affinity and pore forming properties of proteins/peptides - Easy and fast by using solid supported lipid bilayers. Binds a protein to a biological membrane and form parts/domains of this protein a pore after incorporating in a plasma membrane of a cell ? To answer these questions we have developed an easy and quick assay to quantify membrane affinity and to analyse pore formation using TRANSIL®[1] in a micro well plate. In this assay the protein/peptide of interest is incubated with a very well defined single lipid bilayer on a solid support shortly. Separation of the bound/unbound fractions occurs in a low speed centrifugation step and for quantification e.g. a western blot is used. Pore formation will be analysed by ion sensitive fluorescence dyes beneath the lipid bilayer in a fluorescence spectrometer. Taking advantage of the broad spectrum of possible lipid compositions, the identification of binding partners or binding conditions of a protein will be accessible. Our examples include gramicidin A, alpha hemolysin, hrpZ and MRP. Literature [1] Loidl-Stahlhofen et al., Solid-Supported Biomolecules on Modified Silica Surfaces A Tool for Fast Physiocochemical Characterization and High-Throughput Screening. 2001, Adv. Mater. 13, 1829 - 1834 contact: Dr. Joachim Nöller NIMBUS Biotechnologie GmbH [email protected] Eilenburger Str. 4 04317 Leipzig (Germany)

Lars-Oliver Klotz, Helmut Sies, Peter Schröder (-)-Epicatechin Is Taken up by Cells and Protects against Peroxynitrite-Induced Oxidation and Nitration. The dietary flavanol (-)-epicatechin (EC) protects against nitration and oxidation reactions of the inflammatory mediator peroxynitrite in both hydrophilic and hydrophobic environments (1,2). Nitration and dimerization of tyrosine and tyrosine derivatives are prevented with the highest efficiency. EC may thus be a dietary agent with protective effects, but its bioavailability and cellular uptake are still a matter of debate. Here we demonstrate the ability of murine aortic endothelial cells (MAEC) to remove epicatechin from cell culture media during 3h. Based on these findings we investigated if this EC incubation results in a protection of cells. MAEC were incubated for 1 hour in cell culture media containing different concentrations of EC (0.001 2 mM), washed with PBS and finally treated with peroxynitrite. The cells were strongly protected against peroxynitrite-induced oxidation of intracellular dichlorodihydrofluorescein and nitration of protein tyrosyl residues in a dose-dependent fashion, favoring the protection against tyrosine nitration. The protection was comparable to that achieved with EC present during peroxynitrite treatment. In summary, we here show that EC can be taken up by MAEC and that this uptake results in a protection of the cells against peroxynitrite. Literature (1) Schroeder P., Klotz L.-O., Buchczyk D.P., Sadik C.D., Schewe T., and Sies H. (2001). Biochem. Biophys. Res. Comm. 285, 782-787 (2) Schroeder P., Zhang H., Klotz L.-O., Kalyanaraman B., and Sies H. (2001). Biochem. Biophys. Res. Comm. 289, 1334-1338. contact: Dr. Peter Schröder Heinrich-Heine-Universität Institut für Physiologische Chemie I [email protected] Universitätsstr. 1 40225 Düsseldorf (Deutschland / NRW)

Cornelia Körting, Alexander Froschauer, Reinhold Hanel, Jean-Nicolas Volff, Anne-Marie Veith DMRT Genes and Sex Determination in the Fish Xiphophorous Almost nothing is known about the genetic basis of sex determination in fish. Fishes display many different types of sex determination systems, from hermaphroditism to gonochorism, and from environmental sex determination to genetic sex determination including male and female heterogamety, polyfactorial systems and autosomal influences. In the platyfish Xiphophorus maculatus, the sex-determining region on the sex chromosomes is flanked by two related receptor tyrosine kinase genes, Xmrk and egfr-b. The primary sex-determining gene remains to be identified in the platyfish and in the great majority of other fish species. Some members of the DMRT (Doublesex and MAB-3-related transcription factors) family like doublesex in Drosphila melanogaster, mab-3 in Caenorhabiditis elegans and DMRT1 in vertebrates function in sex determination. In animals, DMRT genes can be considered as an interface between variable, lineage-specific upstream cascades of regulatory genes activated by primary sex-determining signals, and downstream structural genes responsible for sex differentiation. On the other hand, some DMRT genes might function as primary sex-determining genes in certain vertebrate lineages. In order to understand the function and evolution of DMRT genes in sex determination in fish, a platyfish bacterial artificial chromosome (BAC) genomic library was screened under low stringency using a heterologous DMRT probe from the medaka fish Oryzias latipes. Six different groups of DMRT-containing BAC clones potentially containing a total of eight different DMRT genes were isolated. As observed in mammals, medaka and pufferfish, DMRT1 and DMRT2 are physically clustered. Novel fish DMRT genes were identified as well. Interestingly, two ancient DMRT duplicates orthologous to one unique mammalian gene were detected in the platyfish. These duplicates might be remnants of the proposed whole genome duplication having arisen early during fish evolution. Differential evolution of paralogous sex-determining genes in different fish lineages after genome duplication might be involved in the amazing variability of sex-determining systems observed in fish.

contact: Dipl. Anne-Marie Veith Universität Würzburg Physiologische Chemie I [email protected] Am Hubland 97074 Würzburg (Deutschland) additional information Biofuture Research Group "Evolutionary Fish Genomics" Funded by the Biofuture program of the Bundesministerium für Bildung und Forschung

Christian Monnerjahn, Ina Schüttmann, Manfred Konrad A designed variant of Herpes virus thymidine kinase selectively phosphorylates the anticancer and antiviral prodrug ganciclovir The selective phosphorylation of nucleoside analogs by thymidine kinases of the Herpes virus family provides the basis for the treatment of viral infections. The human Herpes simplex virus type 1 thymidine kinase gene (HSV1-TK) is also frequently used together with the guanosine derivate ganciclovir (GCV) in suicide gene therapy, though the enzyme strongly prefers thymidine to GCV as substrate. Here we show that the Equine Herpes Virus type 4 thymidine kinase (EHV4-TK) phosphorylates GCV almost as efficient as thymidine. In order to further increase its substrate selectivity towards GCV, we introduced a point mutation (A143Y) in the nucleoside binding pocket of EHV4-TK. This mutation abolishes the thymidine and thymidylate kinase activity, whereas the phosphorylation rate for GCV is not significantly reduced; also, no competitive inhibition by thymidine was detected. Thus, EHV4-TK (A143Y) is a potent new enzyme variant for any TK/GCV-dependent selection strategy, e.g. in eukaryotic cell transfection screening or cancer suicide gene therapy. contact: Dr. rer. nat. Christian Monnerjahn MPI für Biophysikalische Chemie Abt. Molekulare Genetik [email protected] Am Fassberg 11 37077 Göttingen ( Deutschland)

Carsten Skarke, Helmut Schmidt, Jürgen Liefhold, Gerd Geisslinger, Jörn Lötsch A mathematical model to predict plasma concentrations of morphine and its active metabolite morphine-6-glucuronide in healthy young persons Aim: To develop a mathematical model that describes the plasma concentrations of morphine and its active metabolite morphine-6-glucuronide (M6G) and to test its prediction performance Methods: Population compartmental pharmacokinetic modeling using NONMEM was applied to plasma concentration versus time data of morphine and M6G obtained from eight healthy volunteers (4 men, 4 women, aged 23 – 30 years) after intravenous bolus injection of 5.64 mg morphine base (7.5 mg morphine sulfate) and of 1 mg deuterized M6G. The prediction performance was tested by applying the obtained model to morphine and M6G plasma concentrations available from eight other healthy young subjects. Results: Two models were identified that described the plasma concentration versus time courses of morphine and M6G after administration of morphine. The first model consisted of a standard three-compartment model for morphine, and a standard two-compartment model for M6G, with input into and output from the central compartments. An alternative model assigned the formation of M6G among the first peripheral compartment of morphine and the central compartment of M6G, while morphine and M6G were again described by three and two compartment models, respectively. Both models predicted morphine and M6G plasma concentrations available from an independent study with acceptable accuracy and without bias. Conclusions: Two models were obtained that predict plasma concentrations of morphine and M6G with acceptable accuracy in healthy young volunteers. Acknowledgement: DFG Lo 612/2-1 pharmazentrum frankfurt, Department of Clinical Pharmacology, Johann Wolfgang Goethe-University, Theodor Stern Kai 7, D-60590 Frankfurt, Germany Mundipharma GmbH, Mundipharma-Strasse 2-6, D-65549 Limburg (Lahn), Germany contact: Priv.-Doz. Dr. med Jörn Lötsch Universität Frankfurt pharmazentrum frankfurt, Institut für Klinische Pharmakologie [email protected] Theodor-Stern-Kai 7 60590 Frankfurt (Deutschand)

Thomas Tuschl, Sayda M. Elbashir, Jens Harborth, Kim Bechert, Mariana Lagos-Quintana, Javier Martinez, Abdullah Yalcin, Agnieszka Patkaniowska, Jutta Meyer, Klaus Weber A new world of tiny RNAs - siRNAs and miRNAs Approximately 21-nt RNAs and a group of conserved proteins mediate sequence-specific posttranscriptional silencing in almost all eukaryotic organisms. The source of small guide RNA molecules are (1) long dsRNAs naturally produced by replicating RNA viruses and randomly integrated transposable elements or (2) short RNA stem-loops encoded in the genome. The guide RNAs are similar in size because RNase III Dicer processes these dsRNA molecules. The cellular processing of long dsRNAs gives rise to small interfering RNA duplexes (siRNAs), which act as guide RNAs for sequence-specific mRNA degradation. The characteristics of siRNAs are their specific length as well as their base-paired structure containing a 2-nt 3'-overhang. Guidelines for siRNA-mediated gene-specific silencing will be provided based on extensive application of the technology in somatic mammalian cells. Sequencing of endogenous 21-nt RNAs from total RNA isolated from fly embryos, human HeLa cells, and specific mouse tissues, lead to the identification of a new gene family, now termed microRNA genes. More than 100 novel microRNAs have been identified in our laboratory, some of which are developmentally or tissue-specifically expressed. The orthologues of C. elegans lin-4 RNA, the first characterized member of the miRNA family, were just identified in D. melanogaster and mouse. In contrast to siRNAs, which guide the degradation of their target RNAs, it is believed that miRNAs control translation of yet to be identified target mRNAs. contact: Dr. Thomas Tuschl Max-Planck-Institut für Biophysikalische Chemie [email protected] Am Fassberg 11 37077 Göttingen (Germany)

Nils Hanekop, Susanne Wied, Stefan Petry, Norbert Tennagels, Wendelin Frick, Günter Müller A Receptor for Glycolipid Headgroups Mediates Insulin-mimetic Signaling in Rat Adipocytes The phosphoinositolglycan(-peptide)(PIG-P) portion of glycosylphosphatidylinositol-anchored plasma membrane (GPI) proteins is recognized by a binding protein which is located in high-cholesterol-containing detergent-insoluble glycolipid-enriched raft domains (hcDIGs) of the adipocyte plasma membrane. In isolated rat adipocytes, synthetic PIG molecules induce the redistribution of GPI proteins from hcDIGs to low-cholesterol-containing DIGs (lcDIGs) and concomitantly provoke insulin-mimetic signaling and metabolic action. Using a set of synthetic PIG(-P) derivatives we demonstrate here that their specific binding to hcDIGs and their insulin-mimetic signaling/metabolic activity strictly correlate with respect to (i) translocation of the GPI proteins, Gce1 and 5'-nucleotidase, from hcDIGs to lcDIGs, (ii) dissociation of the non-receptor tyrosine kinase, pp59Lyn, from caveolin residing at DIGs (1), (iii) translocation of pp59Lyn from hcDIGs to lcDIGs, (iv) activation of pp59Lyn, (v) tyrosine phosphorylation of insulin receptor substrate proteins (IRS-1/2)(2), and finally (vi) stimulation of glucose transport. The natural PIG(-P) derived from the carboxy-terminal tripeptide of Gce1, YCN-PIG, exhibits the highest potency followed by combination of the separate peptidylethanolamidyl and PIG constituents. We conclude that efficient cross-talk of PIG(-P) to the insulin signaling cascade requires a receptor protein for PIG(-P) at hcDIGs, which may retain GPI proteins within hcDIGs in the unstimulated state but upon occupancy by soluble PIG(-P) release them for lateral movement to lcDIGs which initiates the DIGs-caveolin-pp59Lyn-IRS pathway (3,4) Literature 1) Schlegel, A., Volonte, D., and Engelmann, J.A. (1999) Cell. Signal. 10, 457-463. 2) Nystrom, F.H., and Quon, M.J. (1999) Cell. Signal. 11, 563-574. 3) Müller, G., Jung, C., Wied, S., Welte, S., and Frick, W. (2001) Biochemistry 40, 14603-14620. 4) Müller, G., Jung, C., Wied, S., Welte, S., Jordan, H., and Frick, W. (2001) Mol. Cell. Biol. 21, 4553-4567. contact: Dr Günter Müller Aventis Pharma Deutschland GmbH DG Metabolic Diseases [email protected] Industriepark Höchst Bldg. H825 65926 Frankfurt am Main (Deutschland)

Albert Zlotnik, Anja Muller, Bernhard Homey A role for Chemokine Receptors in Cancer Metastasis? It has been recognized fo some time that chemokines and their receptors control the traffic of leucocytes in the body. We hypothesized that they may also control the traffic of metastatic cancer cells. To test this hypothesis, we measured the expression of 16 chemokine receptors in breast cancer samples. Our results indicated two findings: Firstly, the expression of these receptors in tumor cells is not random but rather it is a specific pattern, and secondly, two receptors in particular, CXCR4 and CCR7 were expressed at high levels in most samples analyzed.Importantly the ligands of these receptors,CXCL12 and CCL21, respectively, were expressed at high levels in lymph nodes, which are primary metastatic destinations of breast cancer cells. In addition, CXCL12 is also expressed strongly in lung, liver and bone marrow, all common metastatic destinations of breast cancer. These observations strongly suggest that these receptors participate in directing organ specific metastasis. Finally, we sought to obtain proof of principle for this hypothesis by testing the ability of a blocking anti-CXCR4 monclonal antibody in an orthotopic model of breast cancer metastasis in mice. We observed that anti-CXCR4 treated mice survived longer and exhibited sharply less metastatic foci in the lung than mice treated with control antibodies. These observations strongly suggest a role for chemokines and their receptors in organ specific metastasis. We have also analyzed the expression of chemokine receptors in many cancers, and have observed that CXCR4 shows widespread expression in many cancers. These observations suggest that CXCR4 is a target of primary importance in the metastatis of many cancers. contact: Prof. Albert Zlotnik EOS Biotechnology [email protected] San Francisco (USA)

Sabine Groesch, Irmgard Tegeder, Karin Schilling, Ellen Niederberger, Gerd Geisslinger Activation of c-Jun-N-terminal-kinase by R- and S-flurbiprofen results in cell cycle arrest in human colon carcinoma cells The unspecific cyclooxygenase inhibitor S-flurbiprofen and its "inactive" enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in several mouse models as well as in rats. The mechanisms underlying the antiproliferative effects of R- and S-flurbiprofen are unknown. Here we show that both R- and S-flurbiprofen inhibit survival of three colon cancer cell lines which differ in the expression of COX-2 (HCT-15: no COX-2, Caco-2: inducible COX-2 and HT-29: constitutive COX-2) which was accompanied by induction of apoptosis in all three cell lines as indicated by DNAand PARP-cleavage. In addition, R- and S-flurbiprofen treatment resulted in a G1-cell cycle block. These effects were associated with an activation of c-Jun N-terminal kinase (JNK), an increase of the DNA binding activity of the transcription factor AP-1 and downregulation of cyclin D1 expression. Supershift experiments indicated that R- and S-flurbiprofen-induced AP-1 activation was associated with a shift in its Jun-protein composition from c-Jun towards JunB. The latter is known to repress cyclin D1 expression. Inhibition of JNK activity prevented the R- and S-flurbiprofen-induced AP-1 DNA binding activity, the repression of cyclin D1 expression and the G1-cell cycle block. However, JNK inhibition had no effect on flurbiprofen-induced apoptosis. These data suggest that the cell cycle inhibitory effects of R- and S-flurbiprofen are mediated at least in part through activation of JNK and subsequent down-regulation of cyclin D1 whereas R- and S-flurbiprofen-induced apoptosis is largely independent of JNK activation. contact: Dr. rer. nat. Sabine Groesch Klinikum der Johann Wolfgang Goethe-Universität pharmazentrum frankfurt, Institut für Klinische Pharmakologie [email protected] Theodor-Stern-Kai 7, Haus 75, 4.OG 60950 Frankfurt (Deutschland)

Kotb Abdelmohsen, Lars-Oliver Klotz, Helmut Sies Activation of EGF receptor-dependent signaling pathways by alkylating and redox-cycling quinones Various quinones, such as the mitomycins or daunorubicin/doxorubicin, are in use clinically in the therapy of solid cancers. We here demonstrate that quinones not only kill cells but also activate cellular signaling pathways that are known to be responsible for proliferation, such as pathways emanating from the epidermal growth factor receptor (EGFR), in WB-F344 rat liver epithelial cells. Extracellular signal regulated kinases (ERK) 1 and 2 as well as protein kinase B/Akt are activated by both alkylating and redox-cycling quinones, including 2-methyl-1,4-naphthoquinone (menadione; MQ), 2,3-dimethoxy-1,4-naphthoquinone (DMNQ), a pure redox-cycler, and p-benzoquinone (BQ), a strong alkylator. ERK phosphorylation was induced by these quinones and was significantly diminished in the presence of the MEK 1/2 inhibitor U0126 or the EGFR kinase inhibitors AG1478 or compound 56. Similarly, the phosphoinositide-3-kinase (PI3K)/Akt pathway was activated, as demonstrated by Akt phosphorylation in the presence of these quinones and the inhibition thereof by the above EGFR kinase inhibitors as well as the PI3K inhibitors wortmannin and LY294002. Glutathione (GSH) was lowered (20-25%) when cells were treated with 0.05 mM menadione for 15 min, whereas almost no depletion was seen at this time point with DMNQ (0.1 mM) and a much stronger depletion (70-75 %) was observed with BQ. In line with the data for the alkylators MQ and BQ, diethyl maleate, a potent GSH depletor (80-85% depletion within 15 min) was capable of activating ERK and Akt, and AG1478 and compound 56 prevented this effect. In summary, both alkylating and redox-cycling quinones activate EGFR-dependent pathways. While GSH depletion may be a potential mechanism for initiating the activation of the EGFR in the case of the alkylators, a mechanism still has to be defined for the activation of the EGFR by the redox-cycler DMNQ. contact: M.Sc. Kotb Abdelmohsen Heinrich-Heine-Universität Düsseldorf Institut für Physiologische Chemie I [email protected] Universitätsstr. 1 40225 Düsseldorf (Germany) additional information Supported by Deutsche Forschungsgemeinschaft (SFB 575/B4).

Hannemann Frank, Bernhardt Rita, Zearo Silvia Activities of CYP11B isoforms from different species. Cytochromes P450 of the CYP11B family are involved in the final steps of glucocorticoid and mineralocorticoid synthesis. In humans CYP11B1 catalyses the production of cortisol and CYP11B2 the synthesis of aldosterone. In many other mammals similar enzymes were found, which catalyze the same reactions. The alignment of the DNA sequences shows high similarity (between 70% and 95%) among the members of the CYP11B family. But the enzyme activities of the various species are found to be different. For a better understanding of the structural basis of the selectivity of CYP11B- dependent reactions we tried to clone and expressed CYP11B genes from various mammalian species (human, monkey, cow, rat and guinea pig) under the same conditions. For the expression we always used the same pNMT1-TOPO vector. With this vector we transformed S. pombe and then expressed the various CYP11Bs proteins. Because of a recently discovered electron transfer protein in S. pombe this cellular system can also be used as a new model for the comparison of the enzyme activity of CYP11B family members. Together with data from computer modelling of the 3D-structure this system might help to get new information about the relationship between sequence, structure and function of CYP11B proteins. Literature Bernhardt R., Rev Physiol Biochem Pharmacol. 1996;127:137-221. Review. Bureik M., et al., Biochemistry. 2002 Feb 19;41(7):2311-21. Hampf M, et al., Endocr Res. 1996 Nov;22(4):495-9 Nomura M, et al., J Biochem (Tokyo). 1993 Feb;113(2):144-52. Nonaka Y, et al., J Steroid Biochem Mol Biol. 1992 Mar;41(3-8):779-80. Bulow HE, et al., Biochem Biophys Res Commun. 1996 Apr 16;221(2):304-12. Denner K, et al., Endocr Res. 1995 Feb-May;21(1-2):443-8. Bechtel S, et al., Eur J Biochem. 2002 Feb;269(4):1118-27. Belkina NV, et al., J Inorg Biochem. 2001 Dec 15;87(4):197-207. Maundrell K., J Biol Chem. 1990 Jul 5;265(19):10857-64. contact: Diplom Biologin Zearo Silvia Universität des Saarlandes Biochemie [email protected] Im Stadtwald, Gebäude 9, Postfach 151150 66041 Saarbrücken (Deutschland)

Svetlana Tsareva, Florian Corvinus, Bernd Wiederanders, Roland Kaufmann, Edith Pfitzner, Richard Moriggl, Karlheinz Friedrich Activities of oncogenic STAT3 in colorectal cancer cells STATs (Signal Transducers and Activators of Transcription) regulate fundamental cellular processes such as proliferation, differentiation and cell death in response to cytokines and growth factors. STAT3 possesses oncogenic properties and was found aberrantly active in various malignant tumors, especially of hematopoietic origin. We have analyzed about 40 colorectal carcinoma biopsies and observed constitutive STAT3 tyrosine phosphorylation and specific DNA binding activity the vast majority of cases. Recent studies revealed connections between STAT3 and the transcriptional regulation of various proteases with relevance to cancer malignancy (e.g. MMP-1, MMP-7). Therefore, we analyzed mRNA from cancer samples by means of cDNA arrays covering 50 different protease probes. In most tumor tissues investigated, a massive up-regulation of matrix metalloproteinases MMP-1, MMP-3, MMP-7 and MMP-9 was observed in comparison to control tissues from the same patients. These findings lead us to the working hypothesis that STAT3 may contribute to the expression of enzymes with importance for the invasive properties of colorectal tumors. In order to further approach a potential funcional involvement of STAT3 in the malignant transformation of colon epithelium cells, we introduced various STAT3 derivatives into the colon carcinoma cell line HT-29. Heterologous overexpression of STAT3 and activation via the interleukin-6 receptor resulted in accelerated cell proliferation. This effect was more pronounced upon transfection with a constitutively active STAT3 mutant, which in addition elicited an altered protease expression profile. A dominant negative STAT3 variant suppressed these cellular responses. These results demonstrate an important role of STAT3 in the growth control of a colon carcinoma model cell line. contact: Svetlana Tsareva Uni Jena Institut für Biochemie [email protected] Nonnenplan 2 07743 Jena (Deutschland) additional information Affiliation of Author 4: Universitätsklinikum Jena Klinik für Allgemeine und Viszerale Chirurgie Affiliation of Author 5: Georg-Speyer-Haus, Frankfurt a.M. Institut für Biomedizinische Forschung Affiliation of Author 6: Institute of Molecular Pathology (IMP) Wien

Andreas Simm, Sherif Daoud, Christian Casselmann, Rolf-Edgar Silber, Reinhard Schinzel Advanced Glycation Endproducts induced cell signalling in cardiac fibroblasts Cardiovascular diseases are the leading cause of death in the Western World, especially in the elder population. One pathophysiologically important mechanism is myocardial fibrosis in aged hearts which is primarily determined by the response of cardiac fibroblasts. We therefore investigated the activation of primary fibroblasts from hearts of adult rats by Advanced Glycation Endproducts (AGEs). AGEs, products of nonenzymatic glycation of proteins, accumulate in various tissues during ageing and are described to be responsible for the stiffness of the heart and vessels. It was shown that treatment of cardiac fibroblasts with AGEs leads to an activation of different signalling molecules such as the p38MAP-kinase, the extracellular regulated kinases (ERKs), the jun kinase (JNK) as well as transcription factors like ATF-2 and NF-kappaB. In addition, the expression and activation of MMP-2, MMP-9 and MMP-13 was induced, which may be responsible for tissue remodelling followed by fibrosis. contact: PD Dr. Andreas Simm Universität Halle-Wittenberg Klinik für Herz- und Thoraxchirurgie [email protected] Ernst-Grube-Str. 40 06120 Halle (Germany)

Markus Kellner, Patrick Baer, Martin Wehling, Ralf Loesel Aldosterone action in humans -new members of the aldosterone signaling pathwayThe primary action of aldosterone in kidney epithelia is to regulate electrolyte and water homeostasis by activation of epithelial sodium channels (ENaCs). The mechanisms by which aldosterone activates ENaCs are not well understood. It seems likely that some aldosterone-regulated genes which lead to an activation of other signaling cascades, are involved in the activation of ENaCs. Microarray technology was used to identify early genes modulated by aldosterone in primary human renal distal epithelial cells (D-TEC). Radioactively labeled cDNA probes from D-TEC total RNA were hybridized with Clontech's Atlas human 8 K microarrays. Of the more than 8.300 genes analyzed, 56 genes (0.67 % ) were up-regulated and 75 genes (0.90 %) down-regulated within 2 h in a range from 3.9-fold to 0.24-fold in response to 100 nM aldosterone treatment. Some differentially expressed genes were confirmed by competitive RT-PCR. Genes are involved in G-protein signaling, GTP-binding, ras signaling, ion-channels, metabolism, differentation, protein trafficking and transcription. The function of some identified aldosterone regulated genes are unknown, but there is evidence that some genes may be involved in ENaCs activation.

contact: Markus Kellner Klinikum Mannheim Klinische Pharmakologie [email protected] Theodor Kutzer Ufer 1-3 68167 Mannheim (Deutschland)

Carolin Oberle, Astrid Matzke, Silvia Meixner, Ulrich Massing, Harald F. Krug Alkylphosphocholine Analogues induce Apoptosis in Tumour Cells of the Immune System Besides other important functions lipids are involved in the formation of cellular structures, cell protection and energy storage. Moreover, they act as signalling molecules which regulate many biological pathways. Some of them can even provoke apoptosis. In this context we investigated the efficacy of synthetic alkylphosphocholines as potential anti-cancer membrane-affecting drugs. The results obtained so far provided evidence that these compounds induce programmed cell death via cell-receptor signalling(1). Leading to novel therapeutic strategies for cancer treatment the new agents interact with the cell membrane and do not affect the DNA. The data presented here show a cell death inducing capacity for 1-O-phosphocholine-2[S]-O-acetyl-octadecane and 1-O-phosphocholin-2[S]-N-acetyl-octadecane in Jurkat T cells as well as in BJAB-cells. The onset of apoptosis is strongly reduced in FADD- and in Caspase-8-deficient cells. This is valid for all parameters tested: caspase activation, substrate cleavage, DNA laddering and PS-externalisation. The activation of caspases is required for the induction of apoptosis as shown by experiments with specific caspase-inhibitors. Additionally, the formation of a functional DISC seems to be involved as indicated by our results with FADD- and Caspase-8-deficient cells. Literature 1. Matzke, A., Massing U. and Krug, H.F. (2001) Eur. J. Cell Biol., 80: 1 contact: PD Dr. Harald F. Krug Forschungszentrum Karlsruhe GmbH Institute of Toxicology and Genetics [email protected] P.O.Box 3640 76021 Karlsruhe (Germany)

Sabine Werner, Stefan Kuklinski, Brigitte Schmitz Altered O-GlcNAc level after neuronal differentiation of PC12 cells O-linked N-acetylglucosamine (O-GlcNAc) is an ubiquitous and abundant post-translational modification found on many cytosolic proteins, nuclear proteins and also on the cytosolic tail of transmembrane proteins. It is a dynamically regulated modification similar to phosphorylation and plays a role in regulation of cellular processes (Zachara and Hart, 2002). To assess potential cell cycle functions of O-GlcNAc we induced the differentiation of PC12 cells by treatment with nerve growth factor (NGF). The O-GlcNAc level decreased significantly after differentiation. This observation indicates that a high O-GlcNAc level of proteins may be more important for dividing cells than for fully differentiated cells. An increased O-GlcNAc level has also been detected in Alzheimer`s disease (Griffith and Schmitz, 1995), which may be linked to the reactivation of the cell cycle, a process currently discussed to be implicated in the pathogenesis of neurodegenerative diseases like Alzheimer`s disease. Literature Griffith, L and Schmitz, B. (1995), Biochem Biophys Res Commun 213 (2), 424-431 Zachara, N. and Hart G.W. (2002), Chem Rev 102 (2), 431-438 contact: Dipl. Biologin Sabine Werner Uni-Bonn Für Anatomie und Physiologie der Haustiere [email protected] Katzenburgweg 9a 53115 Bonn (Deutschland)

Yvonni Chovolou, Wim Wätjen, Andreas Kampkötter, Regine Kahl ALTERED SENSITIVITY OF TNF-ALPHA OVEREXPRESSING RAT HEPATOMA CELLS AGAINST EXOGENOUS TNF-ALPHA AND OXIDANTS Tumor necrosis factor-alpha (TNF-±) is an inflammatory cytokine that causes cell injury by generating oxidative stress. Since ROS production is assumed to be involved in the TNF-± cytotoxicity TNF-± resistance might be brought by the upregulation of antioxidant enzymes. We therefore assumed that endogenous TNF-±(enTNF-±) would protect hepatoma cells from oxidative stress mediated by exogenous TNF-± or prooxidative compounds. Cells overexpressing TNF-± were protected against toxicity mediated by exTNF-± or the redox cycler doxorubicin. Surprisingly, overexpression of TNF-± into H4IIE cells led to an increased sensitivity towards H2O2. Conversely, co-treatment of hepatoma cells with 20 mM of the catalase inhibitor 3-amino-1,2,3-triazole and 15 ng/ml TNF-± for 24 h protected H4IIE cells against the TNF-±-induced cytotoxicity. The status of several antioxidative enzymes was analysed by Northern blot hybridisation, RT-PCR and biochemical assays. In contrast to other cell systems, we found in our system neither a difference in the basal expression of MnSOD between H4IIE and transfected cells nor an induction of MnSOD after exposure to exTNF-±. However, contrary to our expectations catalase mRNA expression as well as catalase enzyme activity were lower in the TNF-± resistant cells than in H4IIE cells. Exposure to exTNF-± at non-toxic concentrations led to a marked decrease in CAT expression in H4IIE cells but not in the transfected cells. TNF-± is known to induce apoptosis in different cell types. DNA fragmentation assay was used to assess apoptosis after treatment with exTNF-±. We observed DNA fragmentation in H4IIE cells while no DNA fragmentation could be detected in transfected cells. During this apoptosis an increase in caspase-3,-8,-9-like protease activities was observed in H4IIE cells but not in TNF-± overexpressing cells. In conclusion, enTNF-± protects H4IIE cells against exTNF-± and doxorubicin but not against H2O2. These results suggest that catalase and intracellular H2O2 concentration may play an important role in the protection against TNF-±-induced cytotoxicity. contact: Dipl. Biol. Yvonni Chovolou Uni Düsseldorf Toxikologie [email protected] 101007 40001 Düsseldorf (D)

Jens Wulfänger, Tanja Blosz, Alexander Navarrete Santos, Jürgen Langner, Dagmar Riemann Aminopeptidase N/CD13 is associated with Lubrol rafts Prominin is a pentaspan membrane glycoprotein associated with a novel type of cholesterol-based lipid rafts [1], which are located especially in structures protruding from the planar regions of the plasmalemma (microvilli, filopodia). Whereas the ‘classical’ cholesterol-sphingolipid rafts are characterized by their insolubility in the non-ionic detergent Triton X-100 at 4 °C, prominin was found to be completely soluble in this detergent, but can be preserved using Lubrol WX as another non-ionic detergent [2]. Earlier studies of our group showed that the membrane-associated ectoenzyme aminopeptidase N/CD13 is partially located in rafts in monocytes [3] and in rafts and caveolae in fibroblast-like synoviocytes [4]. Now, we show that APN is predominantly associated with Lubrol rafts in myeloid cell lines and fibroblasts. Our results point to a special role for APN in filopodia of cells. Literature [1] Corbeil D et al Traffic 2 (2001) 82-91 [2] Roeper K et al Nature Cell Biol 2 (2000) 582-592 [3] Navarrete Santos A et al Biochem Biophys Res Commun 269 (2000) 143-148 [4] Riemann D et al Biochem J 354 (2001) 47-55 contact: Dipl. biochem. Jens Wulfänger Martin Luther University Halle-Wittenberg Institute of Med. Immunology [email protected] Magdeburger Straße 2 06097 Halle (Germany)

Christoph Hutter, Silke Bergmann, Stefan Burdach, Martin S. Staege Analysis of EWS/Fli-1 induced gene expression in HEK293 cells using DNA-microarrays Ewing tumor (ET) is a systemic malignancy originating from bone or soft tissues and is characterized by aggressive local growth, a high metastatic potential and a high rate of early relapses. The chromosomal translocation t(11;22), which can be detected in over 80% of ET, results in expression of a chimeric transcription factor, EWS/Fli-1. It was shown that EWS/Fli-1 expression is necessary for tumor cell growth. However, the mechanisms of transformation by EWS/Fli-1 are unclear and only a few target genes of EWS/Fli-1 have been identified. In order to investigate the influence of EWS/Fli-1 on gene expression in tumor cells we transiently and stable transfected the human embryonic kidney cell line HEK293 with an expression vector allowing simultaneous expression of EWS/Fli-1 and EGFP (pIRES2- EWS/Fli-1-EGFP) and analysed the gene expression profile of transfected cells using high density DNA-microarrays (Affymetrix HG-U133A). Wild type HEK293 cells and cells transfected with empty pIRES2-EGFP were used as control. We identified approximately 150 genes that were up-regulated in EWS/Fli-1 transfected HEK293 cells by a factor of more than 1.5. In addition, more than 200 genes were down- regulated. 66 genes were expressed exclusively in EWS/Fli-1 transfected cells and were absent in all controls. On the other hand, 15 genes were completely absent in all transfected cells but present in all controls. Up-regulated genes were also found in established ET cell lines and native tumor samples. Among the EWS/Fli-1 regulated genes we found a significant number of genes corresponding to markers of neuronal differentiation. In addition, EWS/Fli-1 induced genes included genes involved in signal transduction, cell cycle regulation, and regulation of gene expression. These candidates may be responsible for the proliferation of ET cells in vitro and in vivo. contact: Dr. rer. nat. Martin S. Staege Martin-Luther-University Halle-Wittenberg Children's Cancer Research Center and Department of Pediatrics [email protected] Weinbergweg 22 06097 Halle (Germany)

Harald F. Krug, Claudia Ball, Silvia Diabate Analysis of oxidative stress due to fly ash and ultrafine particles in macrophages and epithelial cells of the lung Epidemiological studies showed that enhanced levels of particulate air pollution (PM10) are associated with an increased incidence of respiratory and cardiovascular diseases. Evidence indicates that particle-induced oxidative stress in lung cells plays an important role in the induction of inflammatory processes in the lung. This study focuses on the generation of reactive oxygen species (ROS) in lung cells after exposure to fly ash from a municipal waste incinerator plant as a model for realistic air pollution particles and to synthetic ultrafine particles. Rat lung derived alveolar macrophages (NR8383), human THP-1 cells differentiated to macrophages and human bronchial epithelial cells (BEAS-2B) were loaded with the nonfluorescent probe 2´,7´-dichlorodihydrofluorescin (H2DCF) which is oxidized by intra- and extracellular hydrogen peroxide generated after exposure to particles or control stimulants. The fluorescent product dichlorofluorescein (DCF) was monitored using a fluoro-spectrometer. We observed a strong increase of fluorescence intensity due to ROS formation in macrophages as well as in epithelial cells after exposure to fly ash and synthetic ultrafine particles in dependence of the mass concentration. This effect was mainly induced by the water-insoluble fraction of the fly ash and could be reduced in the presence of the antioxidant N-acetylcysteine as well as of the metal chelator desferrioxamine. These results indicate that oxidants produced after exposure to insoluble particles generated by combustion processes likely contribute to proinflammatory activation of lung epithelial cells and macrophages and that metals associated to the particles play an important role in this context. This work was supported by the Ministry of Environment and Traffic, Baden-Württemberg, Germany. contact: Dr. Silvia Diabate Forschungszentrum Karlsruhe Institut für Toxikologie und Genetik [email protected] Herrmann-von-Helmholtz-Platz 1 76344 Eggenstein-Leopoldshafen (Germany)

Albert Sickmann, Marion Herrmann, Joachim Klose, Helmut E. Meyer, Heike Schaefer Analysis of Posttranslational Modifications of alpha-A-Crystallin during Aging of the Eye Lens The eye lens allows to focus pictures on the retina. It consists of two types of cells, epithelial and fiber cells [1]. The lens grows throughout life, maintaining transparency without significant turnover of its densely packed proteins [2]. Mainly the crystallins are responsible for the transparency. They comprise more than 90% of the lens proteins. During aging these proteins are posttranslationally modified, i.e. phosphorylated. We report the identification of different posttranslationally modified alpha-A-Crystallin after 2D-PAGE described by Klose et al. [3] of eye lenses (mus musculus) of different ages. The number of detected protein spots of alpha-A-Crystallin in the gel raised from 3 protein spots in the embryo state to 22 protein spots in the gel of the lens proteins analyzed from the oldest mouse (100 weeks). This let to the assumption that the number of modifications in the alpha-A-Crystallin accumulates with age. Using nano-LC-MS/MS alpha-A-Crystallin could be identified with a sequence coverage 50% in all protein spots. Furthermore, some posttranslational modifications, like phosphorylation of specific serine residues could be detected. A set of these protein spots was also analyzed by MALDI-TOF-MS. With these addditional measurements the data of the LC-MS/MS could be verified and complemented. This work demonstrates that LC-ESI-MS/MS and MALDI-TOF-MS are powerful complementary tools for the identification of posttranslational modifications from single 2D-PAGE proteinspots. Literature [1] Colvis, C. M. et al., Electrophoresis 2000, 21, 2219-2227. [2] Jaenicke, R., Naturwissenschaften 1994, 81, 423-429. [3] Klose, J. et al., Electrophoresis 1995, 16, 1034-1059. contact: Heike Schaefer Ruhr-Universitaet Bochum Medizinisches Proteom-Center [email protected] Universitaetsstr. 150 44780 Bochum (Deutschland)

Klaus Heeg Anti-infective intervention via Toll-like receptors Innate immune cells sense infectious danger through recognition of pathogen associated molecular pattern (PAMP). Toll-like receptors (TLR) play a pivotal role in recognition of PAMP. Although various TLRs with distinct ligand specificities are expressed almost completely and contemporaneous by innate immune cells and although central key signal transduction pathways of TLRs are shared, the innate response pattern is not uniform. In addition central signaling pathways of TLR are interconnected with other signaling pathways especially with JAK/STAT dependent intracellular signals. Cis- and trans-acting signal convergence mechanisms and mutual influence of TLR and cytokine signaling pathways determine the response profile. These multiple signal processing and integration pathways offer the chance to intervene at multiple levels. Knowledge of these complex mechanisms will help to understand the critical events during initiation of host responses during infection that in turn may lead to new strategies to control and overcome infectious disease. contact: Prof. Klaus Heeg Universität Marburg Institute of Medical Microbiology [email protected] Pilgrimstein 2 35037 Marburg (D)

Kurt Engeland Apoptosis induction and cell cycle regulation at the G2/M-checkpoint by p63, p73 and the tumor suppressor p53 The p53 tumor suppressor protein is a main regulator in induction of programmed cell death. It can act as a transcription factor on many genes relevant for checkpoint control during the cell division cycle and for induction of apoptosis. Recently, two p53 homologues, p63 and p73, have been identified. Different from p53, p63 and p73 exist in many splice variants. Some of the resulting proteins can also act as transcription factors, e. g. on genes like the one for the cyclin-dependent kinase inhibitor p21WAF1/CIP1. We have demonstrated that regulators which control transition from the G2 phase of the cell cycle to mitosis are p53 target genes. Among these genes are cyclin B and the gene for the cdc25C phosphatase. p53 and its homologues are very different in their transcriptional properties on G2/M-checkpoint genes and also display a different ability to induce apoptosis. The results can explain contrasting observations from knockout mice deficient in functional p53, p63 or p73 genes. contact: PD Dr. Kurt Engeland Universität Leipzig Max-Bürger-Forschungszentrum [email protected] Johannisallee 30 04103 Leipzig (Germany)

Daniela Maennel Appropriate TNF production - a decisive factor in sepsis Tumor necrosis factor (TNF) is considered to be one of the most important inflammatory mediators. In a number of infection models it has been shown that endogenous TNF production is important for survival and exogenously applied TNF exerts protection. Neutralization experiments in a mouse septic peritonitis model induced by cecal ligation and puncture (CLP) demonstrated that TNF is only necessary early after CLP. TNF helps to generate the hyper-inflammatory phase by activating the TNF receptor type 1 to exert antibacterial functions and to localize the septic focus. Mice can be protected from bacterial infections by pretreatment with minute amounts of bacterial lipopolysaccharide (LPS). Using gene knock-out mice we found that LPS-induced protection from CLP mortality required the presence of TNF and TNF receptor type 2. This suggests that activation of the TNF receptor type 2 by LPS-induced endogenous TNF alerts the immune system to an oncoming infection. After the initial hyper-inflammatory phase immediately after CLP a hypo-inflammatory phase develops characterized by a reduced TNF production capacity leading to inadequate immune responses. In this state of immunoparalysis following sublethal CLP, we found mice to be highly susceptible to different kinds of bacterial superinfections. While injection of recombinant TNF was able to overcome this immunoparalysis for a Salmonella superinfection, it was detrimental in case of a superinfection with Listeria. Thus, the impaired TNF production capacity seems to account for some aspects of immunoparalysis but bacteria-specific factors can also sensitize the host for toxic TNF actions. These findings mark TNF as critical factor in prophylaxis, during, and after infection. contact: Prof. Daniela Maennel Universitaet Regensburg Institut fuer Pathologie [email protected] F.-J.-Strauss-Allee 93042 Regensburg (Deutschland)

Michael Spoerner, Guido Steiner, Thomas Prisner, Alfred Wittinghofer, Hans-Robert Kalbitzer Are GTP-analogs really good analogs? For structural and kinetic investigations of Guanine nucleotide binding proteins in the active state different slowly-hydrolyzing GTP-analogs are commonly used. In our group, recently it was shown by 31P NMR, that Ras(wt).GppNHp exists in two different conformations (state 1 and state 2), which are in a dynamical equilibrium(1). Binding of effector-proteins stabilizes state 2. Whereas state 2 seems to be very similar to the effector bound state, structural and kinetic studies indicated a disordered, highly dynamic switch I region for state 1 (2). Here we find similar results for the analog GppCH2p. 31P NMR spectra show also splitting of the Pa and Pß signal in the Ras(wt).GppCH2p complex with similar equilibrium constant and rate constants of exchange. In contrast the GTP-analog GTP³S only shows one conformation if it is bound to Ras protein. Replacing the bound Mg2+ in the active site of Ras by Mn2+ allows highfield-EPR-investigations of the Ras-nucleotide complex. Recently we showed a correlation between different conformations seen by 31P NMR and the coordination of ligands of metal ion bound to Ras.GDP (3). Similar results are obtained for the active state of Ras bound to different GTP-analogs. The two conformational states observed by 31P NMR spectroscopy appear to be closely linked to changes in the coordination sphere of the metal ion. We can show, that GppNHp and GppCH2p change the dynamical behaviour of Ras in the active state by destabilizing the conformation necessary for effector binding defined by the equilibrium constant K = [2]/[1] by a factor of 2. This indicates that the dynamic behaviour of Ras.GTP is highly sensitive to slight modifications in the active site and that this in turn affects interaction of Ras with effectors. Literature (1) Geyer, M., Schweins, T., Herrmann, C., Prisner, T., Wittinghofer, A., and Kalbitzer, H.R. (1996) Biochemistry 35, 10308 (2) Spoerner, M., Herrmann, C., Vetter, I.R., Kalbitzer H.R., and Wittinghofer, A. (2001) Proceedings of the National Academy of Sciences USA 98, 4944 (3) Rohrer, M., Prisner, T.F., Brügmann, O., Käss, H., Spoerner, M., Wittinghofer, A., and Kalbitzer, H.R. (2001) Biochemistry 40, 1884 contact: Michael Spoerner Universität Regensburg Institut für physikalische Biochemie und Biophysik [email protected] 93040 93053 Regensburg (Germany)

R.T. Sauer, R. Burton, J. Kenniston, S. Joshi, D. Wah, I. Levchenko, R. Grant, J. Flynn, S. Neher, T. Baker ATP-dependent protein degradation and unfolding by ClpXP ClpX is a bacterial AAA+ ATPase that serves as the substrate-recognition and protein-unfolding component of the ClpXP protease. Proteomic trapping experiments have identified a large number of new biological substrates for ClpXP. Most of these proteins are recognized by ClpX via C-terminal and/or N-terminal peptide sequences. We are using a wide variety of ssrA-tagged proteins to probe the determinants that dictate whether specific molecules can be unfolded by ClpX and degraded by ClpXP. Many AAA+ ATPases are associated with accessory factors that alter substrate specificity or modulate activity. The structure and function of SspB, a specificity factor that delivers ssrA-tagged substrates to ClpXP, will be discussed. contact: Prof. Robert T. Sauer MIT [email protected] Cambridge (USA)

Ulrike D. Epple, Henning Barth, Michael Thumm Aut10p, Mai1p and Aut5p shed new lights on the molecular mechanisms of autophagy in S.cerevisiae Autophagy is a starvation induced transport pathway delivering cytosolic material for degradation to the lysosome (vacuole). It takes place in all eukaryotic cells and helps the cells to survive periods of nutrient limitation. Autophagy starts at the preautophagosomal compartment, a novel organelle, where several autophagy proteins are localized. From this organelle double-membrane layered transport vesicles (autophagosomes) are formed. Autophagosomes unspecifically enclose parts of the cytoplasm and even organelles like mitochondria, peroxisomes and parts of the ER. When the autophagosomes reach the vacuole their outer membrane-layer fuses with the vacuolar membrane and a still membrane-enclosed autophagic body is released to the vacuolar lumen. Inside the vacuole autophagic bodies are finally lysed and broken down together with their cytosolic content. In a previous genomic approach we identified the novel autophagy protein Aut10p and its homologue Mai1p. Mai1p functions in the cvt-pathway, which is morphologically similar to autophagy, but specifically transports only proAminopeptidase I. Both proteins are peripherally membrane attached and their localization is distinct from known autophagy or cvt-proteins. We here present our novel data about the function of these two proteins. Intravacuolar breakdown of autophagic bodies requires lysis of their surrounding membrane. Intracellular lysis of membranes is a fascinating novel feature of eukaryotic cells and obviously needs tight regulation. Since selective lysis of membranes is crucial during the life cycle of several pathogenes it is also of medical interest. With Aut4p and Aut5p, we identifed two proteins essential for lysis of autophagic bodies. Aut4p is an integral membrane protein of the vacuolar membrane and shares some homologies with permeases. We could demonstrate, that the membrane protein Aut5p is a lipase, which is targeted to the vacuolar lumen at 50 nm vesicles via the multivesicular body (MVB)-pathway. We here present our novel insights in the molecular function of Aut5p. contact: PD Dr. Michael Thumm Uni Stuttgart Inst. f. Biochemie [email protected] Pfaffenwaldring 55 70569 Stuttgart (Deutschland)

Anna Stepczynska, Jakob Troppmair, Thomas Voegel, Monika Poppe, Klaus Schulze-Osthoff, Michael Schwarz Bcl-2 antagonist of cell death (Bad) retains its proapoptotic activity despite proteolytic cleavage by caspase-3 Bcl-2 family member, Bad, promotes death by neutralising the protective effect of Bcl-Xl and Bcl-2. The phosphorylation status of Bad determines whether the molecule is sequestered in cytosol by 14-3-3 protein family members or whether it dimerizes with Bcl-xL or Bcl-2 and thereby releases block on cytochrome c release. We demonstrate that overexpression of Bad in MCF7 cell line is sufficient to trigger cytochrome c release and sensitizes the MCF7/casp-3 cells to staurosporine induced apoptosis. We detect cleavage of endogenous Bad during CD95 and staurosporine-induced apoptosis in Jurkat cell and identify the SATD sequence in murine Bad as caspase-3 substrate site. According to our analysis of apoptotic nuclei and cell morphology cleavage of Bad at this site does not improve Bad death promoting activity. Since we observe that overexpression of truncated Bad can induce formation of apoptotic nuclei in vivo, we support view that death promoting ability-of Bad relies mainly on the intact BH3 domain that enables heterodimerization among Bcl-2 family members and is not subjected to the proteolytic control of caspases. contact: Dr.rer.nat. Anna Stepczynska University of Tübingen Pharmacology and Toxicology [email protected] Wilhelmstr. 56 72074 Tübingen (Germany) additional information J.T. is at the Inst. of Medical Radiation and Cell Research, University of Würzburg T.V is at the Dept. of Experimental Dermatology, University of Münster M.P. is at the Div. of Apoptosis and Cell Death, University of Münster K.S. is at the Dept. of Immunology and Cell Biology, University of Münster

Otto-Erich Brodde, Rainer Büscher, Kirsten Leineweber, Heike Bruck Beta-adrenoceptor polymorphisms: does altered in vitro function predict in vivo effects? ²1- and ²2-Adrenoceptors (AR) are polymorphic. ²1-AR: There is a Gly389Arg polymorphism of the ²1-AR with the Gly389 exhibiting reduced in vitro responsiveness upon agonist-induced stimulation. To study this in vivo we assessed exercise-induced increase in heart rate and shortening of heart rate-corrected duration of electromechanical systole (QS2c - a measure of inotropism) in 12 volunteers homozygous for Gly389 and 12 volunteers homozygous for Arg389. In both groups, exercise caused nearly identical increases in heart rate and shortening of QS2c, indicating that the differential responsiveness observed in vitro is not of major functional importance in vivo. ²2-AR: There are at least three polymorphisms of the ²2-AR: the Arg16Gly and the Gln27Glu polymorphisms differ from the wild-type (WT, Arg16, Gln27) ²2-AR in vitro in their susceptibility to agonist induced downregulation. We studied in 16 WT-volunteers, 6 volunteers homozygous for Glu27 and 6 volunteers homozygous for Gly16, the effects of 2-week treatment with 3 x 5 mg/day p.o. terbutaline on terbutaline infusion-induced increases in heart rate and shortening of QS2c. In all three groups oral terbutaline-treatment reduced terbutaline infusion-induced increases in heart rate and shortening of QS2c to a similar extent; time-course experiments, however, revealed that desensitization occurred faster in WT- and Gly16 volunteers than in Glu27 volunteers. Finally, there is a Thr164Ile polymorphism of the ²2-AR with Ile164 exhibiting reduced in vitro responsiveness upon agonist-induced stimulation. We studied in 12 WT-volunteers and in 6 volunteers heterozygous for the Ile164 polymorphism the effects of terbutaline infusion on heart rate and QS2c. In the Ile164 volunteers increases in heart rate and shortening of QS2c induced by the terbutaline infusion were significantly (pMEK>ERKs) is central in the control of cell growth, cell differentiation and cell survival. The fidelity of signaling and the spatio-temporal activation are key determinants for generating precise biological responses. The fidelity is ensured by scaffold proteins, a sort of protein kinase “insulators” and by specific docking sites. The duration and the intensity of the response are in part controlled by the compartmentalization of the signaling molecules. Growth factors promote rapid nuclear translocation and persistent activation of p42/p44 MAP kinases (ERKs) during the entire G1 period with an extinction during S-phase. These features are exquisitely well controlled by the temporal induction of the MAP kinase phosphatases, MKP1-3. MKP1 and 2 induction is strictly controlled by the activation of the MAP kinase module providing evidence for an auto-regulatory mechanism. This negative regulatory loop is further enhanced by the capacity of p42/p44 MAPK to phosphorylate MKP1 and 2. This action reduces the degradation rate of MKPs through the ubiquitin-proteasomal system. Whereas the two upstream kinases of the module, Raf and MEK remain cytoplasmic, p42/p44 MAPKs, anchored to MEK in the cytoplasm of resting cells, rapidly translocate to the nucleus upon mitogenic stimulation. This process is rapid, reversible, and controlled by the strict activation of the MAPK cascade. Following long term MAPK stimulation, p42/p44 MAPKs progressively accumulate in the nucleus in an inactive form. Therefore we propose that the nucleus represents a site for MAPK action, sequestration and signal termination. Finally, results with mice invalidated for each of the p42/p44 MAPK isoforms, will be presented and discussed in regard to the spatio-temporal control. contact: Doctor Jacques POUYSSEGUR CNRS UMR6543 - Centre A. Lacassagne Institute of Signaling, Developmental Biology and Cancer Research [email protected] 33 Avenue valombrose 06189 Nice (France)

Nishith Gupta, Katrin Lehmann, Otmar Asperger P450non System from Acinetobacter sp. EB104: A Unique Bacterial Alkane Hydroxylase P450non system from Acinetobacter sp. EB104 represents the only bacterial alkane monooxygenase for which all components: P450, ferredoxin (Fd) and ferredoxin reductase (FdR) are available as isolated proteins. Thus, this system represents a preferable model to study the principles for cell-free oxidation of very hydrophobic and chemically inert substances. Recent cloning confirmed the isolated system identity and revealed, compared to other bacterial P450s, a prolonged N-terminus containing an amphipathic alpha-helix. Optical biosensor-based protein-protein interaction studies demonstrated the predominant formation of binary Fd/FdR and Fd/P450 complexes by electrostatic interactions. The P450 itself, oppositely to other bacterial P450s, tends to aggregate in the absence of detergents possibly attributing to the role of lipids for its functional or structural integrity. In accordance with optical spectral data, P450 preparations exhibit predominantly low-spin EPR spectra (g = 2.6, 2.26, 1.84) at 60°K with only weak high-spin signals (g = 4.0, 6.05). The equilibrium can be shifted by addition of lipids and potential substrates. Functional activity of reconstituted systems is distinctively increased by Triton X-100 or rhamnolipids. P450non shows high regio-selectivity for terminal C-atoms independently of chain-length. Oxidation of alkyl chains results in corresponding carboxylic acids indicating a multi-functional enzyme. contact: PD Dr Otmar Asperger Universität Leipzig Institut für Biochemie [email protected] Talstraße 33 04103 Leipzig (D)

Kent Söe, Hella Hartmann, Holger Stephan, Bernhard Schlott, Tinna Stevnsner*, Frank Grosse p53 is involved in the repair of human topoisomerase I cleavage complexes In the past few years it has repeatedly been suggested that p53 and human topoisomerase I (htopoI) play a joint role in the response to genotoxic stress. This was supported by the finding that htopoI forms so called "cleavage complexes" in the vicinity of UV-lesions in a p53 dependent manner in vivo. It was shown that p53 physically interacts with htopoI cleavage complexes in vivo. Therefore, it was proposed that p53 and htopoI cooperate in the process of htopoI-induced repair and/or apoptosis. Recently, we have shown that a htopoI cleavage complex can be recognized by a second htopoI molecule which in turn removes the first covalently bound htopoI molecule and creates an about 13 nucleotides long single-stranded gap (Søe et al. 2001, Nucleic Acids Res. 29: 3195-203). This "double cleavage" reaction may provide an entry site for a subsequent repair event by DNA recombination. Here we show that p53 stimulates both the DNA relaxation activity as well as the double cleavage reaction by at least a factor of six. Stimulation of htopoI is mediated by the central part of htopoI and is equally well pronounced by p53 from human, murine, and bovine origins. Also, we present evidence that the p53-stimulated htopoI double cleavage reaction may represent the initiation event for a repair trial by DNA recombination, at least in vitro. Therefore, we suggest that p53 is stimulation of htopoI may play a role in the repair of DNA and - if running out of control - in the induction of genomic instability. Literature Søe et al. 2001, Nucleic Acids Res. 29: 3195-203 contact: Dr Kent Söe Institut für Molekulare Biotechnologie [email protected] Beutenbergstrasse 11 07745 Jena (Deutschland) additional information *Danish Center for Molecular Gerontology, Department of Molecular and Structural Biology, University of Aarhus, Denmark

Agapios Sachinidis, Jürgen Hescheler PDGF-BB induces selective differentiation of mouse embryonic stem cells to cardiac cells Embryonic stem cells (ES) cells isolated from the inner mass of the early mammalian blastocyst-stage embryo are fully pluripotent and can serve as in vitro model for in vivo differentiation. In order to identify growth factors promoting cardiac development we establish a foetal calf serum (FCS)-free medium. The embryonic stem cells of the murine D3 cell line was used in the study. ES cells were grown as spheroidal aggregates (embryoid bodies; EBs) in hanging drops for 2 days and then cultured in suspension for additional 2 days. Four days after aggregation, embryoid bodies were plated on gelatine-coated 48 well-microwell plate (10 EBs per well). After 24 h, 20%FCS containing Dulbecco´s-modified Eagle´s medium (DMEM) was replaced with 0.2%FCS/DMEM. Following 24 h, serum replacement medium (SRM) (DMEM containing 5 µg/ml insulin, 5 µg/ml transferrin and 1 mg/ml bovine serum albumin) or/and PDGF-BB (50 ng/ml) were added to the medium. After 3 days of addition, cardiomyocytes appeared by spontaneously contracting cell clusters in the plated EBs. Contracting cell clusters in the EBs were regular counted under the microscope. After 10 days EBs were lysed, proteins were separated in a 7.5% SDS-PAGE and cardiac specific myosin heavy chain a and b (cMHCa/b) was detected by the enhanced chemiluminescence Western blotting. Stimulation of the EBs with SRM or PDGF-BB caused an increase of the percent of the number of beating EBs from 13±6 (0.2%FCS/DMEM) to 58±10 or 46±9%, respectively (mean±SEM, n=5). A parallel 6-fold and 3-fold increase of cMHCa/b was observed, respectively. Stimulation of the EBs with PDGF-BB in the presence of SRM resulted in a 260±61 (Mean±SEM, n=3) enhancement of the SRM-induced increase of cMHCa/b (=100%). A parallel increase of beating EBs from 80 to 100% was observed. Applying a new differentiation protocol without we were able to promote selective cardiac differentiation of ES cells by insulin and PDGF-BB. Much more important, this protocol allows identification of different growth factors that promote selective differentiation of ES cells to cardiac cells. contact: Prof. Dr. rer.nat. Agapios Sachinidis Universität zu Köln Zentrum für Physiologie und Pathophysiologie [email protected] Robert-Koch-Straße 39 50931 Köln (Deutschland)

Robert Seckler, Monika Walter Pectate Lyase from Bacillus subtilis (BsPel): Mutations in the asparagine ladder cause a temperature-sensitive folding phenotype Pectate lyases are microbial extracellular enzymes that are important during plant pathogenesis. The enzymes cleave a-1,4 linked galacturonic acid by a beta-elimination. Calcium is essential for the enzymatic activity. The main structural building block of pectate lyases is a right-handed paralllel beta-helix. The structure of pectate lyase from Bacillus subtilis (BsPel) has been solved in complex with calcium (Pickersgill et al. 1994). A characteristic feature of parallel beta-helices is the occurence of extensive stacks of polar and hydrophobic side chains filling the interior of the protein core. In Bacillus subtilis pectate lyase (BsPel), three types of side chain stacks are observed: an aliphatic stack, an aromatic stack and an asparagine ladder. Such an asparagine ladder is a conserved feature of many monomeric beta-helices. It is discussed to be critical for stability and may consitute a nucleus for folding (Jenkins et. al. 1998). The effect of disruption of the asparagine ladder was tested in this mutagenesis study. The middle asparagine (Asn271) of the ladder was selected and exchanged for various residues (Arg, Thr, Leu, Pro, Asp, Glu, Gly). All mutants were expressed at 25°C as soluble proteins, although with significantly reduced yields. The specific activity of the mutants was comparable to that of the wild type. The mutants N271T and N271A were selected for a detailed study of their stability and folding in comparison to the wild type protein. Unfolding of BsPel is not freely reversible and transition curves display apparent hysteresis, connencted with a dramatic decrease of refolding rates at intermediate denaturant concentrations. Refolding, as measured by fluorescence, is at least biphasic with the slowest phase coinciding with reactivation. Refolding of both mutants also follows a biexponential time course. At 10°C and pH 7 the folding rate constants of both mutants were identical within experimental error and very similar to wild type. At pH 7.5 and 25°C, however, refolding of both mutants was very slow even at low guanidinium chloride concentrations and there were considerable differences between the two mutants. Reactivation of both mutants was only possible up to 30°C, whereas the wild type was able to refold up to 40°C. The temperature-sensitive folding phenotype of the mutants was even more pronounced at higher pH. Unfolding of both mutants under these conditions (25°C, pH 7.5) was identical and only 2-fold faster than unfolding of the wild type. Thus, the disruption of the asparagine ladder does not appear to drastically destabilize the native structure of BsPel, but the mutations destabilize an essential folding intermediate formed in the fast kinetic phase of refolding. Literature Jenkins, J., O. Mayans, and R. Pickersgill. 1998. Structure and evolution of parallel beta-helix proteins. J Struct Biol. 122:236-46. Pickersgill, R., J. Jenkins, G. Harris, W. Nasser, and J. Robert-Baudouy. 1994. The structure of Bacillus subtilis pectate lyase in complex with calcium. Nat Struct Biol. 1:717-23. contact: Monika Walter Uni-Potsdam Physikalische Biochemie [email protected] Karl-Liebknecht Str. 24-25 / Haus 25 14476 Golm (Germany)

Monika Walter Pectate Lyase from Bacillus subtilis (BsPel): Mutations in the asparagine ladder cause a temperature-sensitive folding phenotype Pectate lyases are microbial extracellular enzymes that are important during plant pathogenesis. The enzymes cleave a-1,4 linked galacturonic acid by a b-elimination. Calcium is essential for the enzymatic activity. The main structural building block of pectate lyases is a right-handed paralllel b-helix. The structure of pectate lyase from Bacillus subtilis (BsPel) has been solved in complex with calcium (Pickersgill et al. 1994). A characteristic feature of parallel b-helices is the occurence of extensive stacks of polar and hydrophobic side chains filling the interior of the protein core. In Bacillus subtilis pectate lyase (BsPel), three types of side chain stacks are observed: an aliphatic stack, an aromatic stack and an asparagine ladder. Such an asparagine ladder is a conserved feature of many monomeric b-helices. It is discussed to be critical for stability and may consitute a nucleus for folding (Jenkins et. al. 1998). The effect of disruption of the asparagine ladder was tested in this mutagenesis study. The middle asparagine (Asn271) of the ladder was selected and exchanged for various residues (Arg, Thr, Leu, Pro, Asp, Glu, Gly). All mutants were expressed at 25°C as soluble proteins, although with significantly reduced yields. The specific activity of the mutants was comparable to that of the wild type. The mutants N271T and N271A were selected for a detailed study of their stability and folding in comparison to the wild type protein. Unfolding of BsPel is not freely reversible and transition curves display apparent hysteresis, connencted with a dramatic decrease of refolding rates at intermediate denaturant concentrations. Refolding, as measured by fluorescence, is at least biphasic with the slowest phase coinciding with reactivation. Refolding of both mutants also follows a biexponential time course. At 10°C and pH 7 the folding rate constants of both mutants were identical within experimental error and very similar to wild type. At pH 7.5 and 25°C, however, refolding of both mutants was very slow even at low guanidinium chloride concentrations and there were considerable differences between the two mutants. Reactivation of both mutants was only possible up to 30°C, whereas the wild type was able to refold up to 40°C. The temperature-sensitive folding phenotype of the mutants was even more pronounced at higher pH. Unfolding of both mutants under these conditions (25°C, pH 7.5) was identical and only 2-fold faster than unfolding of the wild type. Thus, the disruption of the asparagine ladder does not appear to drastically destabilize the native structure of BsPel, but the mutations destabilize an essential folding intermediate formed in the fast kinetic phase of refolding. Literature Jenkins, J., O. Mayans, and R. Pickersgill. 1998. Structure and evolution of parallel beta-helix proteins. J Struct Biol. 122:236-46. Pickersgill, R., J. Jenkins, G. Harris, W. Nasser, and J. Robert-Baudouy. 1994. The structure of Bacillus subtilis pectate lyase in complex with calcium. Nat Struct Biol. 1:717-23. contact: Monika Walter Uni-Potsdam Physikalische Biochemie [email protected] Karl_liebknechtstrasse 24-25 / Haus 25 14476 Golm (Germany)

Julian Arbuckle Pharmacogenetic based clinical trials to improve drug development Much has been written over the last few years regarding the potential impact of pharmacogenetics (PGx) on drug development and commercialisation. PGx offers much to pharmaceutical companies, regulators, payers, prescribers and patients, however exactly how PGx will make a significant impact on medical practise is still up for debate and will be a topic of this discussion. It should be noted that, by applying the advancements in SNP mapping, genotyping, statistical analysis and bioinformatics to clinical data (safety and/or efficacy), PGx will provide insights into why differential efficacy and idiosyncratic adverse events may occur. The recently published PGx proof of concept study by Hetherington et al investigating hypersensitivity reaction (HSR) to abacavir, a reverse- transcriptase inhibitor that is used to treat Human Immunodeficiency Virus (HIV), shows that it is possible to identify a pattern of chromosomal regions that are associated with occurrence of HSR in a small number of patients. The talk will focus on the abacavir data, as well as the collection of PGx data in clinical trials, so as to provide a picture of how personalised medicine might evolve across the development pipeline.

Literature Hetherington, S et al. Genetic variations in HLA-B region and hypersensitivity reactions to abacavir. Lancet 359:1121-1122 (2002) Roses, A. Genome-based pharmacogenetics and the pharmaceutical industry. Nature reviews 1, 541-549 (2002) Lesko, L.J. and Woodcock, J. Pharmacogenomic-guided drug development: regulatory perspective. Pharmacogenomics J. 2, 101-103 (2002) contact: Mr Julian Arbuckle GlaxoSmithKline [email protected] Greenford UB8 0H6 Middlesex (GB)

Dietmar Pfeifer, Martina Kaufmann, Bernd Fiebich, Olivia Spleiss Pharmacogenetic profiling with Biochips: an innovative concept to improve the efficiency of clinical trials Due to their highly parallel nature, DNA chips are valuable tools for the simultaneous detection of known Single Nucleotide Polymorphisms (SNPs) in numerous genes. In the field of pharmacogenetics, SNPs with a well-described genotype-phenotype correlation in several drug metabolizing enzymes are known for quite a while, but their parallel detection was quite laborious in the past. GeneScan’s Pharm-O-Kin Chip was designed to meet the needs of Clinical research and Pharma customers as it presents a fast and reliable method to detect SNPs in several genes of pharmacogenetic relevance. In total, 39 polymorphisms from CYP2D6, CYP2C9 and CYP2C19 as well as NAT2 and MDR1 are represented on the chip, thus covering important enzymes from drug metabolism as well as a transporter protein known to play a role in the bioavailability of administered drugs. The high quality genotyping data of the Pharm-O-Kin Chip will be of great benefit for clinical trials in the future, as all the relevant polymorphisms known to result in variant phenotypes will be determined in one assay. This allows a better design of clinical trials in terms of minimizing the risks for probands and, in addition, will help to save costs in the drug development process. Literature Brockmoeller J., Kirchheiner J., Meisel C., Roots I. Pharmacogenetic diagnostics of cytochrome P450 polymorphisms in clinical drug development and in drug treatment. Pharmacogenomics 1(2): 125-151, 2000 Sachse C., Brockmoeller J., Bauer S., Roots I. Cytochrome P450 2D6 Variants in a Caucasian Population: Allele Frequencies and Phenotypic Consequences. Am.J.Hum.Genet. 60:284-295, 1997 Murphy M.P., Beaman M.E., Scott C., Cayouette M., Benson L., Morris D.M., Polli J.W. Prospective CYP 2D6 genotyping as an exclusion criterion for enrollment of a phase III clinical trial. Pharmacogenetics, 10:583-590, 2000 contact: Dr. Dietmar Pfeifer GeneScan Europe AG BU Diagnostics [email protected] Engesserstr.4 79108 Freiburg (Germany) additional information Adresse Dr. Bernd L. Fiebich Universitaetsklinik fuer Psychiatrie und Psychosomatik Abteilung fuer Psychiatrie und Psychotherapie mit Poliklinik Hauptstrasse 5, D-79104 Freiburg T. +49 761 270-6898/6501 Fax -6917

Ivar Roots, Julia Kirchheiner, Jürgen Brockmöller, Christian Meisel, Kersti Oselin, Thomas Gerloff Pharmacogenetic studies on drug-metabolizing cytochromes P450 and possibilities for individual drug therapy Drug metabolism and elimination is largely determined by functionally relevant cytochrome P450 genetic variants the most important of which are the polymorphic enzymes CYP2C9, CYP2C19, and CYP2D6. We studied the impact of the CYP2C9 polymorphisms on the pharmacokinetics of characteristic substrates in healthy volunteers. In carriers of the CYP2C9 *3/*3 genotype, clearances were one third (celecoxib, glibenclamide), one half (ibuprofen), and one sixth (tolbutamide) of those of the wild-type. Large inter-individual differences in pharmacokinetics are not necessarily reflected in pharmacodynamics, as tolbutamide-mediated insulin secretion did not differ between CYP2C9 genotypes, at least not in healthy volunteers. Many antipsychotics are metabolized by CYP2D6 and CYP2C19. From published studies with either patients or healthy volunteers, we derived dose-related parameters (clearance, AUC, trough concentrations) in order to calculate genotype-adapted dosages which are based on the principles of bioequivalence (1). Doses for carriers of the deficiency (PM) were often 50% or less of those in wild-type. PMs may have high plasma concentrations of drugs and run the risk of adverse effects and toxicity. Two percent of the German population carry a gene dublication of CYP2D6 and run the risk of therapeutic failure. Genotype-adapted dosage could reduce these risks, especially when the drug has a narrow therapeutic window. Literature 1) Kirchheiner J et al. Acta Psychiatr Scand 2001; 104: 173-92 contact: Prof. Dr. med. Ivar Roots Universitätsklinikum Charité, Humboldt-Universität zu Berlin Institut für Klinische Pharmakologie [email protected] Schumannstr. 20/21 10098 Berlin (D) additional information 3) Jürgen Brockmöller, Prof. Dr. med. Georg August Universität Göttingen Abteilung für Klinische Pharmakologie Robert-Koch-Str. 40 37075 Göttingen

Heyo K. Kroemer, Werner Siegmund, Ingolf Cascorbi Pharmacogenetics in the Cardiovascular Field The influence of genetic factors on disposition and action of cardiovascular drugs is increasingly recognized. During the absorption process polymorphisms in drug transporting proteins may alter bioavailability (eg enhanced bioavailability of digoxin is associated with mutations in the MDR-1 gene). Cytochrome P450 mediated metabolism can be affected by genetic polymorphisms since a number of antiarrhythmics and beta-blocking drugs are metabolized by enzymes such as CYP2D6 or CYP2C9. Patients with low enzyme activity may suffer from side effects in particular during early stages of drug treatment (for example when beta-blockers such as metoprolol or carvedilol are used for treatment of chronic heart failure). Finally, various genetic polymorphisms in adrenergic receptors have been demonstrated to be of functional relevance. contact: Prof. Heyo K. Kroemer University of Greifswald Peter Holtz Research Center of Pharmacology and Experimental Therapeutics / Department of Pharmacology [email protected] Fr. Löfflerstr. 23d 17487 Greifswald (Germany)

Susanne Haufe, Ines Schäffner, Renate Ulbrich-Hofmann Phospholipase D from cabbage: Calcium ions essential for activity destabilize the protein conformation Although Phospholipase D (PLD) has gained much attention in basic research as well as in industrial application, knowledge on its molecular structure and catalytic mechanism is still restricted. Recently, two isoenzymes from cabbage (PLD1 and PLD2) have been expressed in E. coli (1), providing the basis for conformational studies on this enzyme. In this paper the structure and stability of PLD2 (92.1 kDa) has been investigated by CD and fluorescence spectroscopy. Since PLD from cabbage is known to need noncytoplasmic calcium concentrations for optimum activity (40 mM), the effect of Ca2+ ions on the spectra and conformational transitions has been included. CD spectra show that the secondary structure of the enzyme mainly consists of beta-sheets. In the presence of 40 mM CaCl2, no alterations in the CD and fluorescence spectra are observed. The fluorescence signals analyzed in the presence of guanidinium chloride (0 to 8 M) or urea (0 to 10 M) indicate a complex mechanism of unfolding with reversible and irreversible steps. Surprisingly, thermal transition reveals that the stability of PLD2 is decreased in the presence of calcium ions. This finding is conform with the observation that the activity of PLD2 can be best preserved in calcium-free buffers. Calcium ions seem to induce a more flexible (active) but less stable protein conformation. Literature 1) Schäffner, I., Rücknagel, K. P., Mansfeld, J., Ulbrich-Hofmann, R. (2002). Eur. J. Lipid. Sci. Technol. 104, 79-87 contact: Dipl.-Biochem. Susanne Haufe Martin-Luther-Universität Halle-Wittenberg Institut für Biotechnologie [email protected] Kurt-Mothes-Straße 3 06120 Halle (Saale) (Deutschland)

Suisheng Zhang, Carsten Köhler, Frank Grosse PHYSICAL INTERACTION BETWEEN NUCLEAR DNA HELICASE II AND WRN HELICASE STIMULATES WRN´S EXONUCLEASE ACTIVITY The progeriatric Werner syndrome is characterized by a molecular defect of the WRN gene. WRN codes for a 3´-5´-directed DNA helicase that contains an additional 3´-5´ exonucelase activity at its N-terminus. Here we show that WRN helicase forms a physical complex with another intranuclear helicase, i.e. nuclear DNA helicase (NDH II), which is also known as RNA helicase A. Moreover, the earlier described NDH II-copurifying enzyme NDH I was identical to WRN helicase. NDH II and WRN copurify over several chromatographic steps, such as Bio-Rex 70, DEAE Sepharose and phosphocellulose. A high salt elution fraction from the latter column did not only contain NDH II and WRN helicase but also DNA polymerase delta and RPA. This suggests that both WRN helicase and NDH II may be involved in DNA replication and/or DNA repair. NDH II and WRN helicase hardly showed a synergistic effect upon the unwinding of a 3´-tailed double-stranded DNA. However such an effect may be overridden by the strong 3´-5´ exonuclease activty of WRN´s helicase. Indeed, NDH II increased WRN´s exonuclease activity when the unwinding activity was halted by omitting a necessary nucleotide cofactor. These data are in agreement with a model where NDH II unwinds DNA, most likely in the course of transcription. Whenever NDH II is arrested, it may recruit WRN helicase, RPA, and DNA polymerase delta to perform DNA repair synthesis. contact: Dr. Suisheng Zhang Institut für Molekulare Biotechnologie - Jena szhang@ Beutenbergstraße 11 07745 JENA (Deutschland)

Sabine Körber, Sandra Barth, Ulrich Schmidt, Christiane Jäger, Constanze Günther, Christoph Parthier, Gerald Böhm Polyomavirus-like particles as a system for gene therapy and drug delivery

Recent advances in molecular biology have led to the development of new therapeutic strategies to address genetic and other severe disorders. A major limit in clinical applications is the inability of the drugs to be targeted to the appropriate site in the body and subsequently to enter cells. Often, drugs exhibit significant side effects, or their effect is not focused at the intended site of action. Here we describe a novel system for the intracellular delivery of therapeutic agents such as DNA, PNA (peptide nucleic acids), peptides, proteins, and small molecule drugs using recombinant polyomavirus-like particles as universal carriers. In order to achieve maximum functionality of the particles, some of the subunits which form the protein shell are produced as protein chimeras. They contain useful or necessary functions such as cell-type specific targeting modules, domains for endosomal release, for intracellular targeting of the drug and for immune system escape. The modules used for the fusion constructs are taken from natural sources, and the respective fusion proteins between the protein shell and the modules are designed using computer-aided protein structure prediction and modelling before their recombinant production.

Literature Krausewicz,N.; Cox, C.; Soeda, E.; Clark, B.; Rayner, S.; Griffin, BE.: Sustained ex vivo and in vivo transfer of a reporter gene using polyoma virus pseudocapsids. Gene Therapy 2000; 7: 1094-1102. Schmidt, U.; Kenklies, J.; Rudolph, R.; Böhm,G.: Site-specific fluoescence labelling of recombinatn polyomavirus-like particles. Biol. Chem. 1999; 380. 397-401. Schmidt, U.; Rudolph, R.; Böhm, G.:Mechanism of assembly of recombinant murine polyomavirus-like particles. J. Virol. Feb.2000; 1658-1662. Guenter, C; Schmidt, U.; Rudolph, R.; Böhm,G.: Protein and peptide delivery via engineered polyomavirus-like particles. FASEB J. 2001; 1646-8. contact: Dr. Sabine Körber AGCT ProGenomics AG [email protected] Weinbergweg 22 06120 Halle (Germany)

Walter-Vesely Seb. Meister Polyvinylnucleobases: Synthesis, characterization and applications in nanotechnology Artificial homopolymeric polyvinylnucleobases with a polyvinyl backbone mimic native RNA, show the ability to hybridize to their complementary native nucleic acid strands, forming so-called "semiplastic" double- and triple-stranded RNA analogues. Many of such pure and peptide complexed more nuclease-resistant nucleic acid arrangements with better immunomodulatory activities have been developed. But their structure-phase behaviour as well as their structure-function relationship remain open problems. The studies of physico-chemical and biological behaviour of these compounds are basic goals of our approach. AFM-studies of surface immobilization of the first artificial nucleobase compound via thiol-gold reaction or silylation procedure show formation of uniform layers of molecules. We polymerized the artificial strands on gold and silicon wafers using chemical methods of chain extension. The wafer fixation of these biologically active and self-organizing structures stimulates new applications for "regenerable" nucleic acid drugs and nucleic acid computing. Literature S. Hoffmann, W. Witkowski in: Mesomorphic order in polymers and polymerization in liquid crystalline media (A. Blumstein, ed.) ACS-Symp. Ser. 74 (1978) 178-236. B. Glück, W.-V. Meister, W. Witkowski, A. Stelzner, S. Hoffmann, J. Basic Microbiol. 28 (1988) 501-507. W.-V. Meister, S. Lindau, A.L. Hauser, Ch. Bohley, U. Gromann, St. Naumann, M. Madre, L. Kovalenko, G. Bischoff, R. Zhuk, S. Hoffmann, Journal of Biomolecular Structure & Dynamics 18(3) (2000) 385-392. W.-V. Meister, S. Lindau, Anton L. Hauser, E. Birch-Hirschfeld, J. Reinert, K. Friese, C. Bohley, S.I. Kargov, U. Gromann, S. Hoffmann, Surface and Interface Analysis 33(2) (2002) 137-145. contact: Dr. Walter-Vesely Seb. Meister BioMera@web POBox 900-239 (HPA) 06054 Halle /Saale (Germany)

Wim Wätjen, Yvonni Chovolou, Petra Niering, Andreas Kampkötter, Gudrun Aussel, Quynh-Hoa Tran-Thi, Regine Kahl PRO- AND ANTIAPOPTOTIC EFFECTS OF QUERCETIN AND FISETIN IN H4IIE-CELLS: IMPLICATION OF OXIDATIVE STRESS Flavonoids are polyphenolic compounds that occur ubiquitously in vegetable foods. Epidemiological studies have made it likely that they could protect against various stages of the cancer process probably due to their antioxidative properties. We used rat H4IIE hepatoma cells to investigate the effects of quercetin and fisetin against H2O2-induced DNA strand breaks (comet assay), H2O2-induced cytotoxicity (neutral red assay) H2O2-induced apoptosis (DNA-ladder formation, caspase-3 activation) and oxidation potential of H2O2 (DCF fluorescence) in comparison to five structurally related flavonoids. Great differences were found among these substances: Low concentrations of fisetin and quercetin were protective against DNA strand breaks induced by H2O2, while other flavonoids were less or not effective. Similar differences were found for the protection against H2O2-mediated cytotoxicity. Quercetin or fisetin were also shown to be protective against H2O2-induced DCF fluorescence, MDA formation and apoptosis. We further investigated the intrinsic cellular effects of flavonoids in high concentrations. Great differences in the cytotoxicity were found: quercetin and fisetin were shown to be relatively toxic to H4IIE cells (IC50=50-150 µM) while rutin and catechin up to 1000 µM did not cause any significant effects on cell viability. Quercetin and fisetin induced apoptosis as determined by oligonucleosomal DNA cleavage and caspase-3 activation. Among the other flavonoids, taxifolin and myricetin but not catechin, rutin and morin induced oligonucleosomal DNA cleavage. High concentrations of quercetin and fisetin were able to generate oxidative stress as demonstrated by MDA formation. Additionally we observed the induction of DNA strand breaks by high concentrations of flavonoids: quercetin treatment led to an increase of average comet length, other flavonoids showed only minor effects. In summary, quercetin and fisetin were found to be the most potent agents among the investigated flavonoids in protecting cells against oxidative stress. On the other hand both flavonoids are relatively toxic at higher concentrations and induce apoptosis, DNA strand breaks and formation of ROS. contact: Dr. Wim Wätjen Uni Düsseldorf Toxikologie [email protected] 101007 40001 Düsseldorf (D)

Calin-Aurel Dragan, Rita Bernhardt, Matthias Bureik Production of steroid hormones by fission yeast cells expressing human mitochondrial cytochromes P450 Genetically engineered microorganisms are being increasingly used for the industrial production of complicated chemical compounds such as steroids; however, there have been few reports on the use of the fission yeast Schizosaccharomyces pombe for this purpose. Mitochondrial cytochrome P450 enzymes play a crucial role in the steroid biosynthesis in human adrenals, catalyzing regio- and stereo-specific hydroxylations. Two of these enzymes (CYP11B1 and CYP11B2) catalyze the last steps of cortisol and aldosterone biosynthesis, respectively. We have expressed CYP11B2 in S.pombe using the strong nmt1 promotor and found, that the transformed yeasts show in vivo the inducible ability to convert deoxycortisol to cortisol as well as deoxycorticosterone to aldosterone, resp.. The bioconversion activity of these strains is sufficiently high to consider their use for industrial applications. Although in mammalian cells mitochondrial P450 steroid hydroxylases depend for their activity on an electron transport chain that consists of the two proteins adrenodoxin and adrenodoxin reductase, no coexpression of these proteins is needed for efficient substrate conversion by intact fission yeast cells, since S.pombe contains the adrenodoxin-homologue etp1 (Bureik et al., 2002). In order to improve this bioconversion process, we optimized the reaction conditions and created a fission yeast expression vector (pCAD1) that allows targeted integration and the fusion of a polyhistidine and a Pk-tag, resp. (Dragan et al., submitted). Transformation of fission yeast with the new vector containing the human CYP11B1 yielded a new strain that displays an increase in cortisol production rate by more than two orders of magnitude. Literature Bureik, M., et al., Functional expression of human mitochondrial CYP11B2 in fission yeast and identification of a new internal electron transfer protein, etp1. Biochemistry, 2002. 41(7): p. 2311-21. contact: Dr. Matthias Bureik Universität des Saarlandes FR 8.8 Biochemie [email protected] Im Stadtwald, Geb. 9.2 66041 Saarbrücken (Deutschland) additional information http://bernhardt.biochem.uni-sb.de/

Sladjana Dukic-Stefanovic, Jovana Gasic-Milenkovic, Winnie Deuther-Conrad, Gerald Münch Proinflammatory signal transduction pathways in N11 mouse microglial cell line activated by "Advanced Glycation Endproducts" Deposition of AGE-crosslinked insoluble protein aggregates (amyloids) is characteristic for many degenerative diseases of the elderly including Alzheimer?s disease. Microglial activation and neuroinflammatory processes have been shown to play a supporting role in functional degeneration as well as cell death of neurons. "Advanced Glycation Endproducts? (AGEs) activate specific pro-inflammatory signal transduction pathways, resulting in the up-regulation of cytokines (ILs, TNF-alpha) and inducible nitric oxide synthase (iNOS), possibly by their interaction with AGE-specific receptors [1, 2]. Our goal was to delineate the AGE-activated signal transduction pathways involved in the induction of pro-inflammatory markers (TNF-alpha, NO) in the murine microglial cell line N-11. Chicken egg albumin-AGE (CEA-AGE), used as a model AGE, induces both NO and TNF-alpha production is a time- and dose dependent manner. The AGE receptor, RAGE, appears to be the starting point for both pathways, since both NO and TNF-a production can be inhibited by a neutralising RAGE antibody. NO and TNF-a production are regulated independently, since NOS inhibitors did not decrease TNF-alpha secretion and a neutralising TNF-alpha antibody did not reduce NO production. Inhibition of the MEK and PI3K pathway, but not of the MAPK-p38 and SAPK/JNK pathways, reduced NO and TNF-alpha significantly, suggesting that simultaneous activation of the first two pathways is necessary for the AGE-induced induction of these pro-inflammatory stimuli.

Literature [1] Neumann A, Schinzel R, Palm D, Riederer R, Münch G (1999) High molecular weight hyaluronic acid inhibits advanced glycation endproduct-induced NF-kB activation and cytokine expression. FEBS Lett 453: 283-287. [2] Wong A, Schinzel R, Wiesinger H, Riederer P, Münch G (2001) Anti-inflammatory antioxidants attenuate the expression of inducible nitric oxide synthase mediated by advanced glycation endproducts in murine microglia. Eur J Neurosci 14: 1-8 contact: Sladjana Dukic-Stefanovic IZKF Leipzig Neuroimmulogical cell biology [email protected] Johannisallee 30A 04103 Leipzig (germany)

Sandra Loeppen, Daniela Schneider, Frank Gaunitz, Rolf Gebhardt, Albrecht Buchmann, Michael Schwarz Promotion of hepatocarcinogenesis is mediated by overexpression of glutamine synthetase in beta-catenine-mutated mouse liver tumors Phenobarbital (PB) is an antiepileptic drug which promotes hepatocarcinogenesis in rodents when administered subsequent to an initiating carcinogen like N-nitrosodiethylamine (DEN). In the mouse, the promotional effect of PB on liver tumor development results from a selective stimulation of clonal outgrowth of hepatocytes harboring activating mutations in the b-catenin gene. Since ß-catenin has recently been shown to be involved in the regulation of glutamine synthetase (GS), expression of GS during PB-mediated promotion of mouse hepatocarcinogenesis was investigated. Preneoplastic and neoplastic liver lesions were induced in 6 weeks old male mice by a single injection of 90 µg/g body wt. of DEN and groups of mice were subsequently kept on PB-containing (0.05%) or control diet for 39 weeks. While 90% of liver lesions from PB-treated mice were GS-positive, occupying 27% of liver tissue, only 35% of lesions, occupying 0.9% of liver, demonstrated overexpression of GS in mice not treated with PB. We have previously shown that b-catenin mutations are present in ~80% of liver tumors from PB-treated mice but are absent in liver tumors from mice treated with DEN only. By analyzing a panel of larger liver tumors we now observed that tumors harboring b-catenin mutations were GS-positive, whereas tumors without b-catenin mutations were GS-negative. Similarly, tumors from an additional mouse carcinogenicity experiment, where PB inhibited rather than promoted hepatocarcinogenesis were almost completely GS-negative. These data suggest that promotion of hepatocarcinogenesis by PB confers b-catenin-mutated tumor cells with a selective advantage by upregulation of GS expression. contact: Prof. Dr. Michael Schwarz University of Tübingen Pharmacology and Toxicology [email protected] Wilhelmstr. 56 72074 Tübingen (Germany) additional information Adress of Authors 3 and 4 is:Institut für Biochemie, Universität Leipzig, Liebigstr. 16, 04103 Leipzig, Germany

Johannes Herrmann, Tom Lutz, Stephan Meier, Marc Preuss, Gregor Szyrach, Frank Baumann Protein insertion into the inner membrane of mitochondria The inner membrane of mitochondria contains a large number and variety of proteins. Most of these proteins are synthesized in the cytoplasm and imported into mitochondria. Inner membrane proteins are inserted on different sorting pathways. Monotopic proteins can be inserted following a translocation arrest at the level of the inner membrane. An insertion route from the intermembrane space is also used by polytopic proteins which have not evolved from bacterial homologues. In contrast, we found that polytopic proteins derived from the endosymbiotic ancestors of mitochondria are first completely imported into the mitochondrial matrix and subsequently inserted into the membrane. This insertion process depends on the membrane potential and resembles the insertion of membrane proteins in bacteria in several respects. The same insertion route is used by inner membrane proteins that are synthesized on mitochondrial ribosomes. Membrane integration of these proteins depends on the Oxa1 protein, a component related to bacterial YidC. Oxa1 forms an oligomeric complex in the inner membrane and contains a matrix-exposed domain which has the ability to bind translating mitochondrial ribosomes and is required for efficient insertion of nascent polypeptides. This suggests that cotranslational membrane insertion of mitochondrial translation products is achieved by a physical contact of translation complexes with the OXA translocase in the inner membrane. contact: Dr. Johannes Herrmann Universität München Institut für Physiologische Chemie [email protected] Butenandtstrasse 5 81377 München (Germany)

M. Cristina Cardoso, Wolfgang Derer, Hariharan P. Easwaran, Heinrich Leonhardt Protein therapy: Applications in tissue regeneration and stem cell research Our group is interested in the molecular mechanisms regulating the establishment and maintenance of terminal differentiation and to devise ways to transiently reverse this state to achieve tissue regeneration. We have previously shown that this proliferation arrest is an actively maintained process that can be reversed upon transgenic expression of the simian virus 40 large T antigen (SV40 TAg) [1] or overexpression of the cellular E2F1 transcription regulator in the presence of IGF1 [2]. In an attempt to avoid the hazards of oncogene transfer and the difficulties of regulating transgene expression, we are developing approaches to directly deliver the gene products, i.e., the proteins to these cells. Taking advantage of the intercellular trafficking properties of the HSV-I VP22 protein, we were able to directly deliver proteins to terminally differentiated cells [3].We generated a chimeric VP22-SV40 TAg fusion protein and showed that it could transduce terminally differentiated muscle cells and induce cell cycle reentry [4], opening the way to exploit this novel strategy for tissue regeneration. This protein transduction method allows for the simultaneous delivery of mixtures of regulatory proteins in a dose and time controlled fashion and it is easy to combine with the application of other compounds. We are now trying to optimize this technology for tissue regeneration in vivo and for the expansion of differentiated cells in vitro. Literature 1. Cardoso, M.C., Leonhardt, H. and Nadal-Ginard, B. (1993). Reversal of terminal differentiation and control of DNA replication: cyclin A and cdk2 specifically localise at subnuclear sites of DNA replication. Cell 74, 979-992. 2. von Harsdorf, R., Hauck, L., Mehrhof, F., Wegenka, U., Cardoso, M. C.and Dietz, R. (1999). E2F1 overexpression in cardiomyocytes induces downregulation of p21CIP1 and p27KIP1 and release of active cyclin-dependent kinases in the presence of Insulin-like growth factor I. Circ. Res. 85, 128-136. 3. Derer, W., Easwaran, H. P., Knopf, C. W., Leonhardt, H., and Cardoso, M. C. (1999). Direct protein transfer to terminally differentiated muscle cells. J. Mol. Med. 77, 609-613. 4. Derer, W., Easwaran, H. P., Leonhardt, H., and Cardoso, M. C. (2002). A novel approach to induce cell cycle reentry in terminally differentiated muscle cells. The FASEB J. 16: 132-133. contact: Ph.D. M. Cristina Cardoso Max Delbrück Center for Molecular Medicine [email protected] Wiltbergstr. 50 13125 Berlin (Germany)

Tom Rapoport Protein Transport In and Out of the ER Protein transport across the ER membrane occurs through a protein-conducting channel formed from the heterotrimeric Sec61p complex. The channel itself is passive; it needs to associate with partners that provide the driving force for translocation and determine directionality. Translocation into the ER can occur co- or post-translationally. Recent structural studies have given us a better view of the channel and how it connects with the ribosome during co-translational translocation. Proteins can also be translocated from the ER back into the cytosol (retro-translocation). To study the first steps in retro-translocation we have used cholera toxin. We have found that protein disulfide isomerase (PDI) in the ER lumen disassembles and unfolds the toxin once its A-chain has been cleaved. PDI acts as a redox-driven chaperone: in the reduced state, it binds to the A-chain and in the oxidized state it releases it. We have also started to look at the last step in retro-translocation, the release of proteins from the ER membrane into the cytosol. Our results show that poly-ubiquitination is required for retro-translocation. An AAA ATPase family member, Cdc48p/p97, and its partner proteins Ufd1 and Npl4p, are subsequently involved in extracting proteins out of the membrane. contact: Professor Tom Rapoport HHMI/Harvard Medical School Dept. of Cell Biology [email protected] 240 Longwood Avenue 02115-6091 Boston, MA (USA)

Dietrich Keppler, Yunhai Cui, Jörg König, Anne T. Nies Proteins mediating vectorial transport across plasma membrane domains Transport, at a sufficient rate, of most endogenous substances, drugs, and toxins across cellular membranes is mediated by transport proteins and not by passive diffusion. Transport of substances across the cellular barriers is a vectorial process involving uptake and export proteins. This is exemplified by double-transfected polarized cells stably expressing a human uptake transporter for organic anions in the basolateral membrane together with an ATP-dependent export pump for anionic substances in the apical membrane (1). The cloning, localization, and functional characterization of transporters has improved our understanding for the tissue selectivity of uptake, metabolism, actions, and excretion of drugs, toxins, and endogenous substances (2-5). The uptake of organic anions across the basolateral membrane of human hepatocytes is mediated by organic anion transporters with overlapping substrate specificity, such as the organic anion transporters encoded by the genes SLC21A6 and SLC21A8 (4,6). The export of organic anions, particularly of conjugates of lipophilic substances with glutathione or glucuronate, across the apical membrane is mediated by the ATP-dependent conjugate export pump multidrug resistance protein 2 (MRP2) encoded by the ABCC2 gene (2,3). MRP2 is a 190 kDa-integral membrane glycoprotein which plays a key role in detoxification. Mutations leading to the absence of functional MRP2 protein from the apical membrane cause Dubin-Johnson syndrome in humans with conjugated hyperbilirubinemia (3). Other members of the MRP family of export pumps for organic anions are localized to the basolateral membrane and include MRP5 that has been identified as ATP-dependent export pump for 3'-5'- cyclic GMP (7). Literature 1. Cui, Y., König, J., Keppler, D. (2001) Mol. Pharmacol. 60: 934-943. 2. Büchler, M., König, J., Brom, M., Kartenbeck, J., Spring, H., Horie, T., Keppler, D. (1996) J. Biol. Chem. 271: 15091-15098. 3. König, J., Nies, A.T., Cui, Y., Leier, I., Keppler, D. (1999) Biochim. Biophys. Acta 1461: 377-394. 4. König, J., Cui, Y., Nies, A.T., Keppler, D. (2000) J. Biol. Chem. 275: 23161-23168. 5. Jedlitschky, G. and Keppler, D. (2002) Vitamins and Hormones 64: 153-184. 6. Cui, Y., König, J., Leier, I., Buchholz, U., Keppler, D. (2001) J. Biol. Chem. 276: 9626-9630. 7. Jedlitschky, G., Burchell, B., Keppler, D. (2000) J. Biol. Chem. 275: 30069-30074. contact: Prof. Dietrich Keppler Deutsches Krebsforschungszentrum Tumorbiochemie [email protected] Im Neuenheimer Feld 280 69120 Heidelberg (D)

Ulrike Sattler, Ingrid Koziollek-Drechsler, Danuta Dormann, Christina Zechel Putative role of the nuclear factor NCNF in neuronal differentiation Previously, we cloned a nuclear receptor from a cDNA library of neuronal derivatives of retinoic-acid induced embryonic carcinoma cells. This factor was designated NCNF (neuronal cell nuclear factor), and is identical to the germ cell nuclear factor GCNF (also named RTR), which had been cloned from murine testis. NCNF/GCNF is an orphan receptor which most likely functions by repressing transcription. NCNF/GCNF was proposed to be a regulator of (i) germ cell development and (ii) neurogenesis and/or neuronal differentiation. Moreover, data published by the O'Malley group indicate that NCNF/GCNF is (i) required for maintenance of somatogenesis and posterior development and is (ii) essential for embryonic survival. We are interested in the role of NCNF in neurogenesis. To elucidate whether NCNF/GCNF could play a role in neuronal differentiation, we generated transgenic PCC7-Mz1 embryonic carcinoma cells. The PCC7-Mz1 cell line is a model of embryonic mouse brain development in that a similar pattern of neural phenotypes, i.e. neurons, astroglia, and fibroblasts, is produced in a comparable time frame, following exposure to retinoic acid (RA). Firstly, we stably transfected the uninduced cells with the tetracycline-repressor expressing vector pcDNA6TR (T-Rex system; InVitrogen), resulting in the subclone Mz1-TRep. Retinoic acid-induced neural differentiation of Mz1Trex1 was indistinguishable from the parental PCC7-Mz1 cells. The clone was then stably transfected with pcDNA4TO vectors containing a sense or antisense insertion of a FLAG-tagged NCNF cDNA. Fate decision was not influenced by the stable transfection of the NCNF-FLAG construct per se, since both, sense- and antisense clones differentiated indistinguishable from the parental PCC7-Mz1 cells. Ectopic expression of sense NCNF-FLAG mRNA influenced stem cell morphology and favoured the formation of neurons during the course of RA-induced neuronal differentiation. Neutralization of endogenous NCNF mRNA by the expression of antisense NCNF-FLAG mRNA downregulated MAP2 expression in maturing neurons and changed the pattern of neural phenotypes. In particular, fibroblasts developed much more rapidly and the ratio of neurons to fibroblasts to glial cells differed significantly from the parental PCC7 cell line. This data establishes for the first time that NCNF is indeed involved in RA-induced neural pattern formation and neuron maturation. contact: Dr. Christina Zechel Johannes Gutenberg Universität Mainz Institut für Physiologische Chemie und Pathobiochemie [email protected] Duesberg Weg 6 55099 Mainz (Germany)

Thomas Langer Quality control of membrane proteins in mitochondria Impaired proteolysis of membrane proteins has been recognised over recent years as the basis of many human disorders. Since then, a number of membrane-embedded proteases were identified which are capable of clipping membrane-spanning segments or completely degrading membrane proteins. We are studying the proteolytic system of the inner membrane of mitochondria which consists of several components highly conserved throughout evolution: Firstly, the m- and i-AAA proteases with catalytic sites at opposite membrane surfaces. These ATP-dependent peptidases form a membrane-integrated quality control system and exert crucial regulatory functions during mitochondrial biogenesis. Strong phenotypes are associated with mutations in the corresponding genes in various organisms. Yeast cells lacking both proteases are inviable, whereas inactivation of one of these proteases causes neurodegeneration in humans. Secondly, the prohibitin complex which presumably exerts chaperone activity and modulates the degradation of membrane proteins by the m-AAA protease. Prohibitins affect cellular longevity in yeast and mammals, which might reflect effects of a disturbed stability of membrane proteins on mitochondrial metabolism. Thirdly, the ABC-transporter Mdl1 which exports peptides generated by the m-AAA protease in the matrix space allowing their subsequent release from mitochondria. Fourthly, a novel class of membrane-embedded peptidases which have overlapping activities with the m-AAA protease. Recent experiments on the turnover of a polytopic membrane protein by AAA proteases and this novel peptidase will be discussed. contact: Prof. Dr. Thomas Langer Universität zu Köln Institut für Genetik [email protected] Zülpicher Str. 47 50674 Köln (Germany)

Bernd Moritz, Gudrun Franke, Martin Siemann, Matthias Reuss, Helmut E. Meyer Quantitative Proteomics with Escherichia coli Mathematical formulations of metabolic pathways contain metabolite and protein concentrations as variables [1]. Therefore, the combination of quantitative metabolite pool analysis with mass spectrometric determination of protein concentration ratios is a promising tool for getting a deeper insight into central regulation structures. Proteins with altered expression levels in diseased tissues are possible drug targets, which stresses the importance of quantitative proteomics in medicine. Quantitative proteomics can be done after in vitro or in vivo labelling. In vivo labelling is done by cell cultivation in 15N labelled ammonium-sulfate, whereas in vitro labelling by deuterated N-Acetoxysuccinimid or by tryptic digestion in H218O is carried out after cell lysis. In vivo labelling of microorganisms is an accurate method [2], but in case of tissue samples only error-prone in vitro labelling techniques or gel-spot intensity measurements are applicable. Therefore, a comparison and statistical analysis of these techniques with a relatively simple system like E. coli is a prerequisite for the analysis of tissue samples. Literature [1] Moritz B.et al. (2000), Eur J Biochem. 267(12), 3442-52. [2] Oda Y. et al. (1999), Proc. Natl. Acad. Sci. 96, 6591-96. contact: Dipl.-Chem. Gudrun Franke Ruhr-Universität Bochum Medical Proteom-Center [email protected] Universitätsstr. 150 44780 Bochum (Germany) additional information B. Moritz, H.E. Meyer, Medical Proteom-Center, Ruhr-Universität Bochum, 44780 Bochum, Germany M. Siemann, M. Reuss, Institute of Biochemical Engineering, Universität Stuttgart, 70569 Stuttgart, Germany

Kerstin Schnurr, Dagmar Kupper, Antje Banning, Cordula Müller, Matilde Maiorino, Gaby Böl, Regina Brigelius-Flohé Redox events in IL-1 signaling Reduction-oxidation reactions that generate ROS, including H2O2 and O2·¯, have been identified as mediators in cellular signaling. These reactive oxygen species, however, lack specificity due to their short life time and high reactivity. To interact with the known signaling pathways, ROS must have direct protein targets which alter their function(s) with an altered redox state. It is not likely that such specific reactions occur without any enzymatic catalysis. We chose the model of IL-1 signaling and tested whether: · reactive oxygen species are produced upon treatment of ECV cells · protein thiols are involved in the activation of the IL-1 receptor associated kinase (IRAK) and the activation of NFkB · selenium-dependent glutathione peroxidases may play a role in the regulation of IL-1 signaling. We found that - ECV 304 cells released O2·¯ and H2O2 upon IL-1 treatment. When glutathione peroxidase activities were increased by selenium supplementation H2O2 release was decreased by 50% but not O2·¯ production. This indicates two sources of H2O2, an intracellular one, responding to intracellular GPx activity, and an extracellular one, the dismutation of O2·¯ (1). - Thiol modifying agents inhibited the activation of IRAK as well as of NFkB in EL 4 and ECV cells (2). - In PHGPx-overexpressing ECV cells, the IL-1-induced NFkB-activation was diminished but not the activation of IRAK. The data show that reactive oxygen species, preferentially hydroperoxides, as well as thiol-containing proteins, which have to be in the reduced state, are involved in early and late events of the IL-1 signaling pathway. The regulatory role of PHGPx can be explained by its capability to reduce hydroperoxides or to make use of hydroperoxides for the modification of protein thiols. Literature (1) Tolando, R., Jovanovic, A., Brigelius-Flohé, R., Ursini, F. and Maiorino, M. (2000) Selenium supplementation reveals two distinct sites of H2O2 production following treatment with pro-inflammatory cytokines. Free Radic. Biol. Med. 28, 979-986. (2) Tewes, F., Böl, G.-F. and Brigelius-Flohé, R. (1997) Thiol modulation inhibits the IL-1-mediated activation of an IL-1 receptor associated protein kinase and NFkB. Eur. J. Immunol. 27, 3015-3021 contact: Prof . Dr. Regina Brigelius-Flohé German Institute of Human Nutrition Dept. Vitamins and Atherosclerosis [email protected] Arthur-Scheunert Allee 144-116 14558 Bergholz-Rehbrücke (Germany) additional information Address Matilde Maiorino: Dept Biochemistry, University of Padua, I-35121 Padua, Italy

Arne Holmgren Redox regulation by thioredoxin and glutaredoxin systems Thioredoxin (Trx) together with thioredoxin reductase (TrxR) and NADPH comprise the thioredoxin system (1). Thioredoxins have a redox-active disulfide/dithiol and are reduced by selenium-dependent thioredoxin reductases with a Gly-Cys-Sec-Gly active site. Thioredoxins in the cytosol, mitochondria or extracellularly are the cells major disulfide reductases required for the control of redox potential and signalling by thiol redox control. Trx is induced by oxidative and other stresses and operates in defence against reactive oxygen species by being the electron donor for thioredoxin peroxidases and methionine sulfoxide reductases. Thioredoxin controls the activity of numerous transcription factors including NFkB or AP1 directly or via Ref 1. Recent results regarding the structure-function of Trx and TrxR will be presented as well as the reactivity of the antioxidant drug ebselen, which recently was shown to target Trx as well as TrxR (2). Glutaredoxin Grx) catalyzes disulfide oxidoreductions involving glutathione (GSH) and the glutaredoxin system comprises Grx, GSH, glutathione reductase and NADPH. Glutaredoxins which have a classical Cys-Pro-Tyr-Cys active site and a binding site for GSH are required for GSH to operate in thiol-dependent reductions like the synthesis of deoxyribonucleotides by ribonucleotide reductase. Glutaredoxins play a specific role in redox regulation via reverible glutathionylation of proteins where both the synthesis and degradation of the mixed disulfide with glutathione is catalysed by multiple dithiol (CPYC) or monothiol (CXXS) glutaredoxins (3). Literature 1.Arner, ESJ and Holmgren, A. (2000), Eur. J. Biochem.,267,6102-6109. 2.Zhao,R., Masayasu, H. and Holmgren, A. (2002) Proc. Natl. Acad. Sci. USA, 99, 8579-8584. 3. Lundberg, M. et al. (2002) J. Biol. Chem. 276, 26269-26275. contact: Dr. Arne Holmgren Karolinska Institut Medical Nobel Institute for Biochemistry [email protected] 171 77 Stockholm (S)

Margarita Fuhrmann, Kurt Racké, Folker Wenzel Reduced NO synthesis during halothane exposure in stimulated rat alveolar macrophages In stimulated alveolar macrophages (AM) arginine is utilized by inducible nitric oxide synthase (iNOS) to produce nitric oxide (NO) as effector and signal molecule. Because during inhalative anesthesia AM are directly exposed to the applied volatile anesthetics in the present experiments the influence of halothane on NO synthesis in rat NR8383 AM was studied. Freshly resuspended NR8383 cells (0.5 Mio. cells/well) were cultured in the absence or presence of lipopolysaccharides (LPS, 1-100 ng/ml, 20h) as well as halothane (2%). LPS induced nitrite accumulation in the culture medium, as a measure of NO synthesis, was reduced by about 30% at all LPS concentrations in the presence of halothane. Halothane attenuated the LPS induced marked increase in iNOS mRNA expression, as analyzed by RT-PCR (iNOS mRNA/β-actin mRNA: controls, 1.8±0.5; LPS, 97.1±26.4; LPS+halothane, 23.6±10.6; p < 0.05, Student´s t-test). In addition, a possible direct effect of halothane on iNOS activity previously induced by LPS exposure for 20h was determined. In these experiments the extracellular accumulation of [3H]-L-citrulline during 1h incubation with [3H] L-arginine in the absence and presence of halothane amounted to 1358.9±105.2 and to 1008.6±57.3 DPM*1000/mg, respectively (p < 0.05, Student´s t-test). In conclusion, halothane reduces nitric oxide synthesis in LPS stimulated alveolar macrophages by inhibition of both iNOS mRNA expression and iNOS activity. contact: Dr. Folker Wenzel Bonn Institut für Pharmakologie und Toxikologie [email protected] Reuterstr. 2B 53113 Bonn (Germany)

Marcel Eugster, Ulla Grauschopf Refolding and Characterization of the N-terminal extracellular fragment of the Pituitary Adenylate-Cyclase Activating Peptide Receptor The structure of class B G-protein coupled receptors (GPCRs) is comprised of seven membrane-spanning helices and an N-terminal extracellular segment which is involved in ligand binding. Members of this receptor class bind peptides of intermediate length such as parathyroid hormone (PTH), secretin, glucagon-like-peptide (GLP-1) and pituitary adenylate cyclase activating peptide (PACAP). The N-terminal segment contains essential cysteine residues, which form disulfide bonds in the native structure. The disulfide pattern of the N-terminal segment of the PTH-receptor which is formed by 6 cysteine residues was elucidated by expression and oxidative refolding of the N-terminal ligand binding domain from E. coli inclusion bodies. A distinct linkage of the cysteine residues was determined. The same pattern was later also found in the human GLP1 receptor. This indicates a common and conserved disulfide pattern of class B GPCRs. Besides receptors, which contain 6 cysteine residues in their extracellular ligand binding domain, there exist also members of class B GPCRs with 7 cysteine residues in the N-terminal segment and one additional cysteine residue in the first extracellular loop. Constructs of the N-terminal domain of human pituitary adenylate cyclase activating peptide (PACAP) receptor which belongs to this group were designed. The receptor fragments were expressed in Escherichia coli in form of inclusion bodies. After oxidative refolding and purification, we demonstrated that the protein fragments form well-defined stable domains. Ligand binding was proved by fluorescence titration. Refolded receptor fragments were able to bind PACAP27 with a dissociation constant in the nanomolar range. Further we are about to crystallize the ligand binding receptor fragments and to elucidate the disulfide pattern. Literature Beck-Sickinger (1996) DDT 1: 502-513 Grauschopf et al. (2000) Biochemistry 39: 8878-8887 Bazarsuren et al. (2002) Biophys. Chem. 96: 305-318 contact: Marcel Eugster ETH Zürich Molekularbiologie und Biophysik [email protected] Gebäude HPK, Hönggerberg 8093 Zürich (Switzerland)

Frank Fürst, Robert Seckler Refolding from Fragments: The case of the proteolytically processed Pea Lectin Pea lectin (Psl) is a dimeric all-&beta-protein. Its backbone is cleaved after folding, yielding two polypeptide chains in one contiguous domain in each subunit. The shorter chain contributes 1 strand to a 7-stranded &beta-sheet and 1 at each end of a 6-stranded sheet. With this intertwined structure, one would not expect the proteolytically processed protein (PslP) to refold after chemical denaturation. We studied its folding and unfolding and compared it to a recombinant unprocessed form structure (PslS, single chain). Methods employed were fluorescence spectroscopy, gel filtration and an activity assay. We show that PslP is able to refold, but with low yields and only within several days, whereas PslS refolds much faster and with better yields. Unfolding is exceedingly slow for both proteins. The first step is dimer dissociation, followed by an unfolding process visible both in fluorescence and far-UV-CD. Mainly because of the slow unfolding, there is a strong apparent hysteresis in the un-/refolding transition of both forms. Thus, no thermodynamic interpretation is possible. The refolding pathway of PslS consists of at least two phases at low concentrations of GdmCl. The two polypeptide chains of PslP have been isolated under denaturing conditions and their behavior under refolding conditions has been characterized. The &beta-chain alone forms a dimer which must have a different topology than the native dimer of &alpha- and &beta-containing subunits. The same species occurs also on the refolding pathway of PslP. Its unfolding and the insertion of the alpha-chain might be the rate-limiting step in PslP folding. contact: Frank Fürst Univ. Potsdam für Biochemie und Biologie [email protected] Karl-Liebknecht-Str. 24-25, Hs. 25 14476 Golm (Deutschland)

Georg Reiser, Mikhail Strokin Regulation of docosahexaenoic and arachidonic acid release in rat astrocytes Balance of docosahexaenoic (DHA) and arachidonic (AA) acids is particularly significant for normal CNS function and during different pathological states in brain. We compared regulation of release of DHA and AA from the rat brain astrocytes, the important suppliers of polyunsaturated fatty acid in neuronal tissue. We show that astrocytes after stimulation with bradykinin, glutamate, thrombine and ATP release [14C]DHA and [3H]AA. Detailed study of ATP-stimulated release revealed involvement of different isoforms of cytosolic PLA2 in release of DHA and AA. Release of DHA was seen as mediated by Ca2+-independent iPLA2. This release could be blocked by selective inhibitor of iPLA2, 4-bromoenol lactone and was not affected by removal of extracellular Ca2+ and by disruption of intracellular Ca2+ release through PLC inhibitor, U73122. Similary, group IV Ca2+-dependent cPLA2 is involved in ATP-stimulated AA release, which could only be reversed by MAFP, a general inhibitor of intracellular PLA2s. This release was abolished by U73122 and by removal of extracellular Ca2+. Dependence on cAMP/PKA pathway was also studied. ATP-stimulated release of DHA could be amplified twofold by both adenylyl cyclase (forskolin) and PKA (dibutyryl-cAMP) activators and was only weakly affected by inhibitors of those enzymes (2,3-dideoxyadenosine and H89, respectively). In contrast to DHA, the release of AA was not attenuated by activators of cAMP/PKA but was almost completely blocked by 2,3-dideoxyadenosine and inhibited by 34% by H89. Our results provide promising possibilities to develop strategies to the control development of a number of pathological states in brain which involve release of PUFA and their metabolic conversion into inflammatory mediators. contact: Dr. Mikhail Strokin Otto-von-Guericke-Universität Magdeburg Med. Fakultät, I. für Neurobiochemie [email protected] Leipzigerstr. 44 39120 Magdeburg (Germany)

Thomas Braun Regulation of muscle cell development and regeneration by growth factors and bHLH proteins A number of different processes control muscle cell development in vivo: (a) cell proliferation, (b) cell differentiation and (c) cell migration which is required for hypaxial muscle formation. While the critical role of bHLH myogenic transcription factors in muscle cell differentiation has been unequivocally demonstrated by gene inactivation experiments in mice the requirement of specific growth factors for muscle cell differentiation in vivo is still an open question. We have studied all three aspects of muscle cell development. (a) To gain insight into control of muscle cell proliferation we have analyzed mutant FGF-2, FGF-5, FGF-6 and FGF-7 strains in wound healing and tissue regeneration assays. Cell transplantation experiments indicated a reduced mobility of transplanted FGF-6 mutant cells in wild-type hosts. In addition, we investigated the role of shh as a muscle cell maintenance factor in shh and sd mutants and in primary organ cultures. (b) To investigate the role of the bHLH mygenic protein Myf-5 for muscle stem cell development and maintenance in adult mice we generated a conditional allele of the myf-5 gene using Cre/loxP technology. (c) Myogenic precursor migration was analyzed in mice by inactivation of the homeobox gene Lbx1 that is expressed in migrating cells and by analysis of epistatic relations of regulators of limb development. contact: Professor Thomas Braun MLU Halle-Wittenberg Institut für Physiologische Chemie [email protected] Hollystr. 1 06097 Halle (Germany)

Bernd Epe Relevance of endogenous oxidative DNA damage: lessons from repair-deficient mice As a consequence of the generation of reactive oxygen species as by-products of the cellular oxygen metabolism, basal steady-state levels of oxidative DNA base modifications such as 7,8-dihydro-8-oxoguanine (8-oxoG) are observed in all types of cells. Since 8-oxoG and other oxidative DNA modifications are known to be mutagenic, it has long been suspected that they might play an important role in carcinogenesis and other diseases.A strategy to verify this hypothesis is to modulate the steady-state levels and determine the resulting effects on spontaneous mutation rates and other consequences. Recently, the generation of mice deficient in Ogg1 protein, the repair glycosylase that initiates the base excision of 8-oxoG, has opened a new field for this type of research. The Ogg1 defect alone increased the steady-state levels of 8-oxoG only moderately due to the presence of an unexpectedly efficient back-up repair mechanism for 8-oxoG. Nevertheless, the spontaneous mutation rates were clearly increased. We now have demonstrated that the Ogg1-independent back-up repair involves the Cockayne syndrome protein Csb. Thus, the steady-state levels of 8-oxoG in the hepatocytes of csb-/-/ogg1-/- double knockout mice were found to be severalfold higher than in ogg1-/- mice and to increase with age. Accordingly, the repair of additional oxidative DNA base modifications induced by photosensitization in immortalized embryonic fibroblasts (mostly 8-oxoG) was only slightly retarded in csb-/- cells, more compromised in ogg1-/- cells, but virtually absent in csb-/-/ogg1-/- cells. The influence of the elevated oxidative DNA base damage in csb-/-/ogg1-/- mice on the incidence of cancer and other diseases remains to be established. contact: Prof Bernd Epe Universität Mainz Institute of Pharmacy [email protected] Staudingerweg 5 55099 Mainz (D)

Jürgen Alves, Dorothee von Witzendorff, Minh-Cam Ha-Thi, Susanne von Pall de Tolna, Gabriele Grabowski Restriction endonuclease EcoRI - fusions for a change of specificity Restriction enzymes very accurately cleave DNA at a specific nucleotide sequence. Recognition of this sequence by amino acid side chains is highly redundant. Therefore, all attempts to change sequence specificity have failed so far. In order to widen the recognition sequence of the restriction endonuclease EcoRI we have inserted short DNA binding domains into the enzyme structure. The tips of the outer or inner arm were chosen as insertion points to place the additional domains above the major groove. One zinc finger of Zif268 or the third repeat of the Myb oncogene were used as DNA binding modules which are likely to have a stable fold. In a site selection assay one of the fusion mutants showed a preference for the sequence AAGGAATTCCTA. Although the EcoRI sequence GAATTC in varying sequence contexts was still cleaved with low velocity. We also created fusions of the two monomers of EcoRI. These fused dimers are even more toxic to cells than the wild type enzyme. This sheds light on a hitherto unrecognized evolutionary advantage of the dimeric state for restriction enzymes. contact: Prof. Jürgen Alves Medizinische Hochschule Hannover Biophysikalische Chemie [email protected] Carl-Neuberg-Str. 1 30625 Hannover (Deutschland)

Frank Czubayko, Achim Aigner Ribozyme-PEI-complexes as ready-to-use reagents for efficient down-regulation of gene expression Ribozymes represent potentially interesting agents in gene therapy since they allow the effective abrogation of the expression of any selected protein. Also, they offer powerful stategies in proteomics / target validation applications. Due to ribozyme instability and poor cellular uptake, however, the use of enzymatically active RNA molecules like hammerhead ribozymes has been severely hampered and largely unsuccessful so far. We have developed a method for protection and cellular delivery of bioactive ribozymes by complexation with a low molecular weight polyethylenimine (PEI). Polyethylenimines are synthetic branched polymers with high cationic charge density which form non-covalent complexes with nucleic acids and have been used as DNA transfection reagents. We show that ribozyme-PEI complexation allows complete stabilization of ribozymes or any RNA against degradation. Upon their highly efficient cellular uptake in vitro, non-toxic PEI-complexed ribozymes display intracellular bioactivity already at low concentrations as demonstrated by down-regulation of two different genes in different tumor cell lines. Moreover, in vivo delivery of PEI-complexed ribozymes against the growth factor pleiotrophin (PTN) results in marked reduction of tumor growth and of intratumoral PTN levels in a mouse xenograft model. Thus, in this study we describe a novel, widely applicable method for exogenous delivery of any bioactive RNA ribozyme in vitro and in vivo without chemical modification. contact: Dr. Achim Aigner Philipps-Universität Marburg Institut für Pharmakologie und Toxikologie [email protected] Karl-von-Frisch-Strasse 1 35033 Marburg (Germany)

Josef H. Wissler RNA as a cytokine [ribokine]: Metalloregulated extracellular eRNA and bioaptamers, the uncovered missing links for cellular communication and signal transduction in an anticipated RNA world of today. Ribosomes, mRNA, tRNA, telomerases, snRNP and ribozymes are evaluated RNA biofunctions in an anticipated RNA world [1]. However, some basic requisites of such world to be alive escaped consideration, so far: Thus, no facts were disclosed, at all, how protocells, the earliest ancestors of contemporary living cells managed intercellular communication, signal reception and transduction. From culture media, cells and wound fluids, sequence-defined structures of highly modified and edited, [Cu,Ca,Zn,Mg,Na,K]- metalloregulated 5'-end phosphorylated extracellular small RNA with cellular, enzymic and bioaptamer functions could be isolated and characterized. Biomolecular switching between functions is operated by Fenton-type OH*-radical redox reactions and metal ions, including RNA modification by formation of some labile adenosine-N1-oxide in transit to isoguanosine nucleotides. As bioaptamers, copper ion-structured eRNA impart novel roles to [e.g. S100-EF-hand] proteins which these have not on their own [2-8]. A physiologic role of endothelial cell receptors for advanced glycosylation end products [RAGE] is shown up in angio-morphogenesis by transduction of signals of RNA bioaptamers complexed by copper ions to calcium ion-regulated S100A12-EF-hand proteins [calgranulins]. By 3D-rapid prototyping molecular modeling methods [9] applied to biomolecular behaviour in the process, eRNA is suggested presented as reformed ssRNA to intracellular RNA-binding RAGE domains as lid of RNA chaperone- shaped S100A12 hexamer assemblies: Activation of the cell is dependent on Ca, Zn/Mg and Cu ions and occurs upon Ca/Cu ion-mediated multivalent binding reactions, S100A12 ligand and RAGE receptor oligomerization to RNA chaperone-shaped S100A12 hexamer assemblies at oligomerized RAGE membrane sites with concomitant formation of cellular asymmetry and polarity centers. As general perspective, ribokines feature RNA as cytokines providing RNA tools for cellular communication. They complement RNA biofunctions of an implemented RNA world in requisite to be alive, at all: No life is possible without communication, since communication is the essence of feedback, homeostasis and defense. That is to be or not to be a living cell and a vital organism. Literature [1] Gesteland, R.F., Cech, T.R. and Atkins, J.F., eds: The RNA World, 2nd ed: The Nature of Modern RNA Suggests a Prebiotic RNA. Cold Spring Harbor Laboratory Press, New York 1999. [2] Wissler, J.H., Logemann, E., Meyer, H.E., Krützfeldt, B., Höckel, M. and Heilmeyer jr., L.M.G.: Structure and function of a monocytic blood vessel morphogen [angiotropin] for angiogenesis in vivo and in vitro: A copper-containing metallo-polyribonucleo-polypeptide as a novel and unique type of monokine. Protides of the Biological Fluids, Peeters, H., ed., Pergamon Press, Oxford 34: 525-536, 1986. [3] Wissler, J.H., Amselgruber, W.M., Schweiger, M. and Logemann, E.: Structure, function and cellular localization of angiotropin, a non-mitogenic leukocytic endothelial cell RNP-morphogen [ribokine] for organoid capillary pattern formation. Mol. Biol. Cell Suppl. 8: 231a, 1997. [4] Wissler, J.H. and Logemann, E.: RNA structure and modification of copper-RNP complex of angiotropin ribokines [non-mitogenic leukocytic endothelial cell morphogens]. FASEB J. 12: A1463, 1998. [5] Wissler, J.H. and Logemann. E.: Ribokines with modified and edited extracellular eRNA: Endogenous metalloregulated copper-ribonucleoprotein cytokines [CuRNP] and their components [S100-EF-hand protein, RNA, precursors] in cellular signal transduction. Biol. Chem. 381: S234 & S246, 2000. [6] Wissler, J.H.: Clonal selection of bioaptamers and diversity of oligonucleotide biolibraries in antibody-independent, antigen-specific delayed-type hypersensitivity [DTH], spongiform encephalopathy [TSE] and related transfer ["infectivity"] reactions. Biol. Chem. 382: S99, 2001. [7] Wissler, J.H.: Engineering of blood vessel patterns by angio-morphogens [angiotropins]: Non-mitogenic copper-ribonucleoprotein cytokines [CuRNP ribokines] with their metalloregulated constituents of RAGE-binding S100-EF-hand proteins and extracellular RNA bioaptamers in vascular remodeling of tissue and angiogenesis in vitro. Materialwiss. Werkstofftech. [Mat. Sci. Eng. Technol.] 32: 984-1008, 2001. [8] Wissler, J.H.: Engineering of capillary patterns in muscle by a nonmitogenic copper-ribonucleoprotein angiomorphogen [angiotropin CuRNP ribokine]. Ann. N.Y. Acad. Sci. 961: 292-297, 2002. [9] Laub, M., Seul, T., Schmachtenberg, E. and Jennissen, H.P.: Molecular modelling of bone morphogenetic protein-2 (BMP-2) by 3D-rapid prototyping. Materialwiss. Werkstofftech. [Mat. Sci. Eng. Technol.] 32: 926-930, 2001. contact: Prof. Dr. Josef H. Wissler ARCONS Applied Research, Education and Didactics [email protected] Postfach 1327 D-61231 Bad Nauheim (Germany)

Jovana Gasic-Milenkovic, Sladjana Dukic-Stefanovic, Winnie Deuther-Conrad, Gerald Münch ß-amyloid peptide potentiates inflammatory responses induced by lipopolysaccharide, interferon-gamma and "advanced glycation endproducts" in a mouse microglia cell line ß-amyloid peptide (Aß) and "Advanced glycation endproducts" (AGEs), protein bound oxidation products of sugars, accumulate in the senile plaques in Alzheimer’s disease (AD) brain. Activated microglial and astroglial cells are associated with the senile plaques, produce free radicals and inflammatory cytokines, and are suggested to accelerate the neurodegenerative process [1]. We hypothesized that Aß needs a pre-stimulated environment to exert its pro-inflammatory potential. Since AGEs accumulate in the AD brain over time, a gradual increase in the level of AGEs may gradually induce a low-level of neurotoxicity and chronic inflammation in the ageing brain [2]. Thus, we tested whether Aß acts as an amplifier of a sub-threshold pro-inflammatory response initiated by exposure to AGEs as the AD-specific physiological stimulus using chicken egg albumin-AGE (CEA-AGEs), or the established stimuli LPS or IFN-gamma. Synthetic Aß (1-40) was used to produce three different samples (large sedimentable Aß aggregates, small non-sedimentable Aß aggregates and Aß-AGE) which were further characterised for fibril content by Thioflavin-T assay and light microscopy. M-CSF, NO and TNF-alpha production were measured as markers of microglial activation. All three Aß preparations could not induce a detectable microglial response on their own. However, the combination of Aß preparations with CEA-AGEs, LPS or IFN-gamma provoked a strong microglial responce. Aß-AGE and the small non-sedimentable Aß aggregates were the most potent co-inducers of the pro-inflammatory response. Thus, we can assume that Aß induces significant microglial activation only in the presence of some other inflammatory stimulus. Literature [1] Wong A, Schinzel R, Wiesinger H, Riederer P, Münch G (2001) Anti-inflammatory antioxidants attenuate the expression of inducible nitric oxide synthase mediated by advanced glycation endproducts in murine microglia. Eur J Neurosci 14: 1-8 [2] Gasic-Milenkovic J, Loske C, Münch G (2001) "AGEing" of a protein – the cytotoxicity of a glycated protein increases with its degree of AGE modification. Z Gerontol Geriatrie 34: 457-60 contact: Dr Jovana Gasic-Milenkovic IZKF Leipzig Neuroimmunological Cell Biology [email protected] Johannisallee 30a 04103 Leipzig (Germany)

Anton Vichalkovski, Karin Kirschner, Kurt Baltensperger, Hartmut Porzig SDF-1 and thrombin signaling in human hematopoietic progenitor cells is regulated by cytokines While much is known about cytokine-activated cellular signaling pathways and their relevance for hematopoietic cell development, the functional role of signals mediated by G protein-coupled receptors (GPCR) that are expressed in these cells has remained unclear. We have now used a chemokine, stromal cell-derived factor 1 (SDF-1), and thrombin, two prototypic endogenous GPCR agonists, to study growth effects and cellular Ca2+ transients in multipotential and in lineage-committed normal hematopoietic progenitors. In multipotential cells, SDF-1 (5-50 nM) caused a 50 to 200% increase in stem cell factor (SCF)-,IL-3or granulocyte- macrophage colony-stimulating factor(GMCSF)-supported DNA synthesis. By contrast, the effects of thrombopoietin (TPO) and of erythropoietin (Epo) were not enhanced. The effect of SDF-1 could be largely blocked by an inhibitor of Rho kinase (Y27632), by inhibitors of protein kinase C and by pertussis toxin (PTX). On the other hand, thrombin (0.5-2 U),in committed erythroid cells, strongly inhibited Epo-stimulated DNA synthesis but did not antagonize the effects of TPO or GMCSF in multipotential cells. The effect of thrombin was blocked by botulinum C3 toxin but not by PTX or Y27632. Both SDF-1 and thrombin caused Ca2+ transients due to cellular release and store-operated influx. In single multipotential cells SDF-1 caused long lasting Ca2+ oscillations. The SDF-1- but not the thrombin-induced changes in cell growth might be correlated to their effects on cellular Ca2+ concentrations. We conclude that GPCR-mediated modulation of hematopoiesis is promoted by cooperative interactions with specific cytokines. contact: Prof. Hartmut Porzig Universität Bern Pharmakologie [email protected] Friedbühlstrasse 49 CH 3010 Bern (Schweiz)

Ernst Petzinger, Kerstin Lischka, Dieter Starke Secretion of intact oligonucleotides by mrp2 into bile As therapeutic antisense tools, oligonucleotides distribute in nearly each tissue with relatively high concentrations in kidney and liver from where excretion into urine and bile occurs. Since most ODNs are polyanions their passage through the cell membrane requires specific translocation processes. To investigate mechanisms involved in hepatic ODN transport, normal mixed backbone phosphodiester/phosphoro-thioate ODNs (n-ODN) and two different bile acid-conjugated mixed backbone ODNs (1BA-ODN, 2BA-ODN) were applied to two different rat strains, normal Wistar rats and Wistar TR(minus)rats. The latter lack a functional mrp2 canalicular export pump. In normal Wistar rats secretion of intact oligonucleotides into bile was observed with a remarkable increase of excretion of the cholic acid-conjugated ODNs. In TR(minus)rats n-ODN excretion was almost nil whereas 2BA-ODN excretion occured, however at low level only. Concomitant application of substrates of mrp2 such as bromosulfophthalein or the synthetic chlorogenic acid derivative S 3025 significantly reduced the biliary appearance of n-ODN and 2BA-ODN in normal Wistar rats. The results indicate that ODNs are secreted via mrp2 into bile. In the absence of mrp2 further excretory transport systems such as the bile salt export pump seem to be relevant for their excretion. Bile acid conjugated ODNs appear to be suitable antisense oligonucleotides with high affinity to the liver. Bile excretion occurs even under restricted conditions of mrp2 dysfunction (e.g. human Dubin-Johnson syndrome). contact: Prof. Dr. Ernst Petzinger Universitaet Giessen Institut f. Pharmakol. und Toxikol. [email protected] Frankfurter Str. 107 35392 Giessen (Deutschland)

Antonios Kyriakopoulos, Barbara Hoppe, Alexandra Graebert, Marcus Kühbacher, Dietrich Behne Selenoproteins in the perinuclear structures of rat kidney Dedt. Molecular Trace Element research in the Life Sciences Hahn- Meitner-Istitut, Glienicker Str.100, 14109 Berlin

The essentiality of the tracer element selenium was shown to be due to the biological function of several selenoproteins. In the selenoproteins identified so far the element is present as selenocysteine and in this form is part of the active site of some enzymes. For the determination of the selenium laves instrumental neutron activation analysis (INAA) via 75Se was used (1) Information on the membrane-bound selenoproteins was obtained by labelling of rats in vivo with 75Se-selenite, subcellular fractionation of the tissue homogenates, separation of the proteins by SDS-PAGE, two-dimensional electrophoresis and selenium detection by autoradiography (2). In this way more than 30 selenium-containing proteins were detected (3). Of those six selenium-containing proteins with the approximately molecular masses of 60kDa, 38kDa, 28kDa, 23-25kDa, 20kDa and 16kDa were detected in the perinuclear structures fraction. The 75Se-labeled protein bands of 60kDa, 38 kDa ,23-25 kDa and 16kDa after treatment with detergents, protein separation by SDS-PAGE and detection of selenium by autoradiography are present in the endoplasmatic reticulum fraction (ER) again. This finding is of great interest regarding the function of the cellular organells. Therefore several experiments have been curried out in order to characterize these selenium-containing proteins.

Literature 1. Behne, D., Kyriakopoulos, A., Weiss-Nowak, C,. Kalcklösch, M. Westphal, C., Gessner, H. Newly found selenium-containing proteins in the tissues of the rat. Biol. Trace Element Res. 55 (1996) 99-110. 2. Kyriakopoulos, A., Röthlein, D., Pfeifer, H., Bertelsmann, H , Behne, D. Detection of small selenium-containing proteins in tissues of the rat. J. Trace Elements. Med. Biol. 14 (2000) 179-183. 3. Behne, D., Scheid, S., Hilmert, H.Gessner, H., Gawlik, D and Kyriakopoulos, A. ombination of neutron activation analysis, tracer techniques, and biochemical methods in the investigation of selenium metabolism. Biological Trace element Research (1990) 439-447. contact: Dr. Antonios Kyriakopoulos HGF Hahn-Meitner-Institut [email protected] Glienicker Str. 100 14109 Berlin (D)

Maria Ludewig*, Carsten Schwarz+, Otmar Asperger*, Svante Pääbo+, Ulrich Hahn* Sequence Analysis of an Acinetobacter Plasmid Carrying the Genes of a P450-Dependent Alkane Monooxygenase In addition to a nonheme iron alkane hydroxylase Acinetobacter sp. EB104 contains a P450-dependent alkane monooxygenase that consists of the haem protein P450non (CYP153) as the terminal oxygenase, a ferredoxin (Fd) and a NADH-dependent ferredoxin reductase (FdR). It was shown by southern hybridisation that the three genes of the P450 system are localised on the native plasmid pAC450 of A. sp. EB104. They are organised in a single operon together with a putative transcription regulator of the AraC/XylS family. To exploit this system for the biotransformation of chemically inert and highly hydrophobic compounds genes of enzymes involved in further steps of alkane oxidation should be identified. Therefore we sequenced the entire plasmid by shot-gun cloning. The draft sequence implies that the P450 operon is part of a 17-kb composite transposon. In the non-transposon region an open reading frame for a protein putatively involved in conjugation was detected suggesting the acquirement of the P450 system by horizontal gene transfer. ORF's with similarities to those of DNA-interacting proteins indicate that the plasmid encodes its own replication assembly. Furthermore, an ORF with similarity to a transport protein for long-chain fatty acids was found, but sequences typical for alcohol or aldehyde dehydrogenases were not detected. contact: PD Dr. Otmar Asperger Universität Leipzig Institut für Biochemie [email protected] Talstr. 33 04103 Leipzig (D) additional information * University of Leipzig, Institute of Biochemistry + Max-Planck-Institute for Evolutionary Anthropology, Leipzig

Alexandra Koch, Annalisa Mancini, Regina Wilms, Teruko Tamura SH2-domain containing inositol 5-phosphatase (SHIP)-1 is a potential regulator of cytoskeletal rearrangements in NGF induced neurite outgrowth Phosphatidylinositol phosphate metabolism plays a crucial role in cytoskeletal rearrangement processes altering cellular properties such as cell shape and motility. To study the role of SH2-domain containing inositol 5-phosphatase (SHIP)-1 we first examined epithelial MDCK cells as a model system. In these cells overexpression of SHIP-1 causes dramatic changes in the cytoskeleton leading to higher cell motility and changes in cell-cell junctions. This phenotype is dependent on SHIP s phosphatase activity. Formation of neurites requires an enormous cytoskeletal rearrangement in the differentiating neuron. Therefore we extended our experiment to PC12 cells as neuronal model system to examine the role of 5 -phosphorylation of phosphatidylinositol phosphates. PC12 cells express SHIP-1 endogenously at very low levels. Overexpression of SHIP-1 in PC12 cells favors flattening of the cell body within 6 hours of treatment with nerve growth factor (NGF) and enhances the following neurite outgrowth throughout 3 days. Whereas within 10 minutes of NGF treatment cells expressing high levels of GFP tagged SHIP-1 form and retract small filopodia faster than controll cells. For the adaptor protein Shc, a well described interaction partner of the NGF receptor TrkA, is a known binding partner of SHIP-1 we expect a possible NGF dependent recruitment of SHIP-1 through Shc and an involvement of SHIP-1 in TrkA mediated cytoskeletal rearrangement. Literature Src homology 2-containing inositol 5-phosphatase 1 binds to the multifunctional docking site of c-Met and potentiates hepatocyte growth factor-induced branching tubulogenesis. Stefan M, Koch A., Mancini A., Mohr A., Weidner K.M., Nieman H., and Tamura T (2001) J. Biol. Chem. 276 (5), 3017-23 The SH2-containing inositol 5-phosphatase (SHIP)-1 is implicated in the control of cell-cell junction and induces dissociation and dispersion of MDCK cells. Mancini A., Koch A., Wilms R., and Tamura T. (2002) Oncogene 21, 1477-84 contact: Dr. rer. nat. Alexandra Koch Medizinische Hochschule Hannover Institut für Physiologische Chemie -OE [email protected] Carl-Neuberg-Str. 1 30623 Hannover (Deutschland)

Thorsten Nürnberger, Dierk Scheel Signal perception and transduction in the plant immune response Immunity of an entire plant species to microbial infection is determined by intertwined layers of defense including both constitutive barriers and inducible reactions. Activation of inducible defense responses is likely based upon recognition of pathogen-associated molecular patterns (PAMP), which bind to plant receptors. We have identified a calcium-dependent cell wall transglutaminase (TGase, EC 2.3.2.13) from the phytopathogenic oomycete, Phytophthora sojae, that triggers defense responses in parsley. Transcripts encoding TGase were expressed and enzyme activity was detectable in 10 Phytophthora species analyzed. A surface-exposed fragment (Pep-13) of this protein, which is sufficient for receptor-mediated activation of pathogen defense, is highly conserved in all Phytophthora TGases. Mutational analysis within the Pep-13 sequence revealed amino acid residues indispensible for both transglutaminase activity and activation of plant defense. Thus, PAMPs (such as Pep-13) recognized by plants show similiarities to those triggering innate immune responses in humans or Drosophila. A 100-kDa monomeric plasma membrane protein constitutes the Pep-13 receptor, which mediates transcriptional activation of pathogenesis-related genes and phytoalexin production in parsley. Ligand-induced receptor activation results in elevated levels of cytoplasmic [Ca2+], which is likely due to influx through plasma membrane [Ca2+] channels. [Ca2+]-dependent production of superoxide anions and activation of MAP kinase cascades are implicated in the activation of transcription factors which mediate the expression of plant defense responses. Literature Nürnberger,T., Nennstiel, D., Jabs, T., Sacks, W.R., Hahlbrock, K., Scheel, D. High-affinity binding of a fungal oligopeptide at parsley plasma membranes triggers multiple defense responses. Cell, 78, 449-460 (1994). Nürnberger, T., Nennstiel, D., Hahlbrock, K., Scheel, D. Covalent cross-linking of the Phytophthora megasperma oligopeptide elicitor to its receptor in parsley membranes. Proc. Natl. Acad. Sci. U.S.A., 92, 2338-2342 (1995). Zimmermann, S., Nürnberger, T., Frachisse, J.-M., Wirtz, W., Guern, J., Hedrich, R., Scheel, D. Receptor-mediated activation of a plant Ca2+-permeable ion channel involved in pathogen defense. Proc. Natl. Acad. Sci. U.S.A. 94, 2751-2755 (1997). Jabs, T., Tschöpe, M., Colling, C., Hahlbrock, K. and Scheel, D. Elicitor-stimulated ion fluxes and O2from the oxidative burst are essential components in triggering defense gene activation and phytoalexin synthesis in parsley. Proc. Natl. Acad. Sci. USA 94, 4800-4805, (1997). Ligterink, W., Kroj, T., zur Nieden, U., Hirt, H. and Scheel, D. Receptor-mediated activation of a MAP kinase in pathogen defense of plants. Science 276, 2054-2057, (1997). Blume, B., Nürnberger, T., Nass, N., Scheel, D. Receptor-mediated rise in cytoplasmic free calcium required for activation of pathogen defense in parsley. Plant Cell, 12, 1425-1440 (2000). Lee, J., Klüsener, B., Tsiamis, G., Stevens, C., Neyt, C., Cornelis, G.R., Panopoulos, N.J., Weiler, E.W., Mansfield, J., Nürnberger, T. HrpZPsph from the plant pathogen Pseudomonas syringae pv. phaseolicola is exported by the type III secretion pathway and forms an ion-conducting pore in vitro. Proc. Natl. Acad. Sci. U.S.A., 98, 289-294 (2001). Lee, J., Klessig, D.F., Nürnberger, T. A harpin binding site to tobacco plasma membranes mediates SIPK-dependent activation of the defense-related gene, HIN1. Plant Cell, 13, 1079-1093 (2001).

Nürnberger, T., Scheel, D. Signal transmission in the plant immune response. Trends Plant Sci. 6, 372-379 (2001). contact: Dr. Thorsten Nürnberger Institut für Pflanzenbiochemie [email protected] Weinberg 3 06120 Halle/Saale (Deutschland)

Julia Lintzel, Inga Franke, Wolfgang Hampe Signal transduction by SorLA, a member of the LDL-receptor family The neuropeptide head activator (HA) promotes proliferation of neuroendocrine and neuronal precursor cells. A pertussis-toxin sensitive G-protein is involved in signaling from the cell surface receptor to induction of mitosis. We identified the single-membrane spanning receptor SorLA (also called LR11) as a mammalian HA receptor. SorLA contains binding domains typical for the low-density lipoprotein receptors and a VPS10 domain, which in the related receptor sortilin binds the neuropeptide neurotensin. SorLA is synthesized as a proreceptor, which is processed to the mature form by a furin-like propeptidase. Pull-down assays using transiently expressed partial SorLA constructs showed that HA interacts with the furin-processed VPS10 domain. A filter binding assay with a radiolabeled HA-analogue revealed a nanomolar affinity of HA to the furin-processed VPS10 domain of SorLA. As in the pull-down assay HA-binding was abolished in the presence of SorLA's propeptide. We therefore used the propeptide as an antagonist to analyze SorLA's function in HA signal transduction. It inhibited HA-stimulated mitosis and proliferation in the human neuronal precursor cell line NT2 and the neuroendocrine cell line BON demonstrating the importance of SorLA for HA function. From these results we conclude that SorLA participates in HA signaling either as a signaling receptor itself or as a coreceptor for another signaling receptor which might belong to the family of G-protein coupled receptors. contact: Dr Wolfgang Hampe Uniklinikum Eppendorf Inst. für Med. Biochemie und Molekularbiologie [email protected] Martinistr. 52 20246 Hamburg (Deutschland)

Manfred Frasch, Hsiu-Hsiang Lee, Patrick Lo, Ingolf Reim, Hideyuki Nagaso SIGNALING AND TRANSCRIPTIONAL CONTROL OF VISCERAL MUSCLE AND HEART DEVELOPMENT IN DROSOPHILA The Drosophila heart, termed dorsal vessel, is a simple contractile tube that is reminiscent of the tubular heart found in early vertebrate embryos. Developmentally, early cardiogenesis in vertebrates and insects also share common features, including the requirement of inductive tissue interactions across germ layers which generate bilateral heart primordia in the lateral mesoderm. The respective linear tubes of the heart and dorsal vessel are then formed upon the fusion of these bilateral fields along the midline. Because there is clear evidence that these morphological and developmental similarities extend to the genetic and molecular level, Drosophila has become a fruitful model to study control mechanisms of early cardiogenesis as well as visceral muscle development. Spatially-restricted inductive signals from the ectoderm, in particular the combination of Dpp (=BMP) and Wingless (=Wnt), are required together with mesodermal activity of the NK homeodomain gene tinman to induce heart progenitors at defined positions within the dorsal mesoderm. Conversely, Dpp + tinman and the absence of Wingless signals are required for visceral muscle formation. Molecular and functional enhancer analysis of heart and visceral muscle progenitor-specific target genes has provided further insight into the mechanisms by which these regulators cooperate molecularly to induce the respective tissues. We will also discuss signaling and transcriptional pathways that determine the diversification of cell types within the developing heart and visceral muscle tissues. Literature Knirr S, Frasch M., Dev Biol. 2001;238:13-26. Zaffran S, Küchler A, Lee HH, Frasch M., Genes Dev. 2001;15:2900-15. Lo PC, Frasch M., Mech Dev. 2001;104:49-60. Xu X, Yin Z, Hudson JB, Ferguson EL, Frasch M., Genes Dev. 1998;12:2354-70. contact: Dr. Manfred Frasch Mount Sinai School of Medicine Dept. of Developmental and Cell Biology [email protected] One Gustave L. Levy Place 10029 New York NY (USA)

Ulrich Laufs Significance of cholesterol-independent effects of statins HMG CoA reductase inhibitiors (statins) have been shown to be effective lipid lowering agents and are able to significantly reduce cardiovascular mortality and morbidity in patients at risk for cardiovascular disease. Recent clinical and experimental data suggest that the benefit of statins may extend beyond their hepatic inhibition of cholesterolsynthesis. The lecture will summarize the current evidence and the molecular mechanisms of the potential direct effects of statins on plaque stability, inflammation, endothelial function, oxidative stress, thrombosis and stroke. Furthermore, the talk will focus on the recently recognised effects of statins on bone marrow derived endothelial progenitor cells. From a research perspective, statins have emerged as a novel tool to study protein isoprenylation, small G protein function, leukocyte activity and progenitor cells. The extra-hepatic properties of statins may have important clinical implications in addition to lowering of serum cholesterol, especially for normocholesterolemic patients with cardiovascular disease and after withdrawal of statin treatment. Literature Werner N, Nickenig G, Laufs U (2002) Pleiotropic effects of HMG-CoA reductase inhibitors. Basic Res Cardiol 97: 105-116 contact: Dr. Ulrich Laufs Universität des Saarlandes Klinik Innere Medizin III [email protected] Geb. 40 66421 Homburg (Germany)

Claudia von Montfort, Lars-Oliver Klotz Singlet Molecular Oxygen Reversibly Inhibits Tyrosine Phosphatases Singlet oxygen, an electronically excited form of oxygen, is a strong oxidant generated in vivo either photochemically, such as under the influence of ultraviolet-A radiation in the presence of cellular photosensitizers, or by stimulated neutrophils in inflammatory processes. Singlet oxygen is capable of activating cellular signal transduction pathways in human skin cells, including p38- and JNK mitogen-activated protein kinases. Regarding the mode of activation of these signaling pathways by oxidants, a commonly held hypothesis is that it is caused by the inhibition of negatively regulating phosphatases by oxidation. Here we demonstrate the inhibition of the two protein tyrosine phosphatases (PTPases) PTP1B and CD45 by chemically generated singlet oxygen. Isolated PTPases were exposed to the 1,4-endoperoxide of naphthalene 1,4-dipropionate (NDPO2) followed by determination of residual enzymatic activity. PTP1B (700 nM) activity was largely abolished by exposure to singlet oxygen steady-state concentrations in the range 4.5 nM. More than 50% activity was restored using DTT. Similarly, 100 nM CD45 were almost totally inhibited by 0.9 nM singlet oxygen, the activity again being restorable more than 70% with DTT, suggesting that mostly disulfides or sulfenic acids, but not sulfinic or sulfonic acids, are generated during oxidation. In addition to oxidation, the inactivation of PTPases may be achieved by arylation of essential cysteine residues. The vitamin K analog menadione, which is known to arylate nucleophilic moieties, is thought to arylate protein tyrosine phosphatases at their active site cysteine. In line with this, CD45 is inhibited by menadione (84%) stronger than by DMNQ, a quinone that cannot arylate. contact: Claudia von Montfort Heinrich-Heine-Universität Düsseldorf Physiologische Chemie 1 [email protected] Universitätsstraße 1 40225 Düsseldorf (Germany) additional information Supported by Deutsche Forschungsgemeinschaft (SFB 503/B1)

Ralph-Peter Elsner Solution structure of the Ras binding domain of AF6 1Guido Steiner, 1Werner Kremer, 1Ralph-Peter Elsner, 2Thomas Linnemann, 2Christian Herrmann, 1Michael Wenzler, 1Gudrun Horn and 1Hans Robert Kalbitzer 1Institut für Biophysik und physikalische Biochemie, Universität Regensburg, 93040 Regensburg 2Max-Planck-Institut für molekulare Physiologie, Otto-Hahn-Str. 11, 44227 Dortmund AF6, a protein with high sequence homology to Drosophila CANOE, was originally identified as a fusion partner of ALL-1 in acute myeloid leucaemia [1]. In 1996, Kuriyama et al. reported the discovery of a Ras binding protein in bovine brain extract [2]. The sequence of this protein with a molecular mass of 180 kDa turned out to be almost identical to human AF6. Thus, AF6 was identified as a putative effector in a Ras modulated signalling cascade. Binding of AF6 was also observed to ZO-1, a protein involved in the formation of tight junctions in epithelial cells [3]. It could be shown that a null mutation in the murine AF6 locus leads to abnormal development of cell-cell-junctions in early embryogenesis, with lethal consequences for the mouse embryo [4]. The amino terminus of AF6 (residues 1-141) adopts a stable fold and was characterized as the minimal Ras binding domain (RBD) [5]. AF6-RBD exhibits Ras binding properties similar to other effectors such as Raf and RalGDS [5]. On the other hand, AF6-RBD is significantly larger than those. Here we present the three dimensional structure of AF6-RBD in solution which was determined by high resolution NMR spectroscopy. The structure showed to be widely identical to other known Ras-binding domains. The fold of AF6 exhibits a five-stranded mixed ²-sheet complemented by two ±-helices, which is known as a ubiquitin ±/²-roll. In comparison to other known RBD structures we observe n-terminal an additional α helix and a longer loop region, thus extending the common fold of RBD´s. Literature [1] Prasad, R., Gu, Y., Alder, H., Nakamura, T., Canaani, O., Saito, H., Huebner, K., Gale, R. P., Nowell, P. C., Kuriyama, K., Miyazaki, Y., Croce, C. M., Canaani, E., (1993) Cancer Res. 53, 5624 [2] Kuriyama, M., Harada, N., Kuroda, S., Yamamoto, T., Nakafuku, M., Iwamatsu, A., Yamamoto, D., Prasad, R., Croce, C., Canaani, E., Kaibuchi, K. (1996) J. Biol. Chem. 271, 607 [3] Yamamoto, T., Harada, N., Kano, K., Taya, S., Canaani, E., Matsuura, Y., Mizoguchi, A., Ide, C., Kaibuchi, K. (1997) J. Cell. Biol. 139, 785 [4] Zhadanov, A., Provance, D. W., Speer, C. A., Coffin, J. D., Goss, D., Blixt, J. A., Reichert, C. M., Mercer, J. A. (1999) Current Biology 9, 880 [5] Linnemann, T., Geyer, M., Jaitner, B. K., Block, C., Kalbitzer, H. R., Wittinghofer, A., Herrmann, C., (1999) J. Biol. Chem. 274, 13556 contact: Ralph-Peter Elsner Uni Regensburg Biophysik/physikalische Biochemie [email protected] Universitätsstr. 31 93053 Regensburg (Deutschland)

Alexander Freiberg, Jan Lengefeld, Matthias Machner, Wolfgang Pfeil, Dirk Heinz, Robert Seckler Stability and Folding of Internalin B - A Biophysical Characterization Internalin B belongs to the internalin family of Listeria monocytogenes and promotes entry by phagocytosis of the Gram-positive pathogen into many normally non-phagocytic cells. The N-terminal fragment of Internalin B (residues 36-211, InlB241) (Schubert et al. 2001) used in this study consists of a leucine-rich repeat domain that is flanked by an N-terminal cap domain. Each single leucine-rich repeat consists of 22 amino-acid residues folded into a short beta-strand and an opposing 310-helix. The beta-strands and helices are connected to each other by turns. Our Internalin B fragment is a monomeric protein, with a molecular mass of 23 kDa and without disulfide bonds. In the present study, we characterized the spectroscopic properties, the stability, and the folding of this leucine-rich repeat protein using different biophysical probes. The pronounced circular dichroism spectra of InlB241 exhibit no fine structure in the near-UV. In contrast, the far-UV CD spectra display a maximum at 185 nm and minima at 219 and 199 nm. The latter may be characteristic for 310-helix. The single tryptophane residue at position 124 exhibits maximum fluorescence around 348 nm, compatible with its solvent exposure. InlB241 shows reversible folding behaviour after chemical denaturation measured by fluorescence at 10 µg/ml. The midpoint of the unfolding transition is at 0.63 M guanidinium chloride. Fitting the guanidinium chloride induced unfolding transition with a two-state model, yielded a free energy of stabilization deltaG (H2O) of - 4.8 ± 0.2 kcal/mol. The relative high denaturation m-value of 7.5 ± 0.3 kcal/(mol M) is indicative of the absence of intermediates and, thus, supports the two-state analysis. By contrast, thermal denaturation is not reversible but results in aggregation. The native structure of InlB241 is stable in the pH range from 6 to 10. Below pH 4, the maximum fluorescence emission wavelength is blue-shifted to 333 nm, suggesting the presence of a collapsed, non-native state. Renaturation kinetics indicate the presence of at least one folding intermediate forming on the stopped-flow time scale. Literature SCHUBERT, W.-D., GÖBEL, G., DIEPHOLZ, M., DARJI, A., KLOER, D., HAIN, T., CHAKRABORTY, T., WEHLAND, J., DOMANN, E., and HEINZ, D. W.: Internalins from the Human Pathogen Listeria monocytogenes Combine Three Distinct Folds into a Contiguous Internalin Domain. J. Mol. Biol. 312, 783-794 (2001) contact: Alexander Freiberg Universität Potsdam Institut für Biochemie und Biologie [email protected] Karl-Liebknecht-Strasse 24 - 25, Haus 25 14476 Golm (Deutschland)

Matthias Gaestel Stress-signalling downstream to p38 MAPK: Essential role of MAPKAP kinase 2 (MK2) MAPKAP kinase 2 (MK2) is regulated via direct phosphorylation through p38 MAP kinase, the central component of a stress-activated kinase cascade. MK2 is mainly localised in the nucleus of non-stressed cells and, as a consequence of activation and conformational change, rapidly exported to the cytoplasm in response to stress (Engel et al., 1998; Neininger et al., 2001). By introducing a targeted mutation into the mouse MK2 gene, we determined the physiological function of this protein kinase in vivo. Mice that lack MK2 show increased stress resistance and survive LPS-induced endotoxic shock. This is caused by an 90 % reduction in the production of tumor necrosis factor (TNF) protein, although the level and stability of TNF mRNA is not reduced and TNF secretion is not altered (Kotlyarov et al., 1999). Interestingly, deletion of the AU-rich element (ARE) in the 3' UTR of the TNF gene rescues the defect in LPS-induced TNF biosynthesis indicating that MK2 is upstream to the ARE at the same genetically defined pathway. We conclude that MK2 is an essential component in the inflammatory response which regulates biosynthesis of TNF at a post-transcriptional level probably by acting through an ARE-dependent mechanism within the cytoplasmic compartment (Neininger et al., 2002). In addition, MK2 is involved in signalling to actin remodelling. Compared with WT cells, MK2-deficient neutrophils show a partial loss of directionality but higher migration speed (Hannigan et al., 2001). MK2-deficient smooth muscle cells and embryonic fibroblasts show also clearly reduced migratory abilities. Interestingly, reintroduction of MK2 catalytic activity is not sufficient for the restoration of the migratory phenotype. In addition to catalytic activity, the proline-rich potential ena/Vasp-EHV1-binding N-terminal region of MK2 is necessary for the rescue of migration (Kotlyarov et al., 2002). These data indicate the requirement of MK2 for both cytokine production and cell migration. Literature Engel, K., Kotlyarov, A., and Gaestel, M. (1998). Leptomycin B-sensitive nuclear export of MAPKAP kinase 2 is regulated by phosphorylation. Embo J 17, 3363-3371. Hannigan, M. O., Zhan, L., Ai, Y., Kotlyarov, A., Gaestel, M., and Huang, C. K. (2001). Abnormal migration phenotype of mitogen-activated protein kinase-activated protein kinase 2(-/-) neutrophils in zigmond chambers containing formyl-methionyl-leucyl-phenylalanine gradients. J Immunol 167, 3953-3961. Kotlyarov, A., Neininger, A., Schubert, C., Eckert, R., Birchmeier, C., Volk, H. D., and Gaestel, M. (1999). MAPKAP kinase 2 is essential for LPS-induced TNF-alpha biosynthesis. Nat Cell Biol 1, 94-97. Kotlyarov, A., Yannoni, Y., Fritz, S., Laass, K., Telliez, J.-B., Pitman, D., Lin, L.-L., and Gaestel, M. (2002). Distinct cellular functions of MK2. Mol Cell Biol 22, in press. Neininger, A., Kontoyiannis, D., Kotlyarov, A., Winzen, R., Eckert, R., Volk, H. D., Holtmann, H., Kollias, G., and Gaestel, M. (2002). MK2 targets AU-rich elements and regulates biosynthesis of tumor necrosis factor and interleukin-6 independently at different post-transcriptional levels. J Biol Chem 277, 3065-3068. Neininger, A., Thielemann, H., and Gaestel, M. (2001). FRET-based detection of different conformations of MK2. EMBO Rep 2, 703-708.

contact: Prof Matthias Gaestel MHH Biochemie [email protected] Carl-Neuberg-Str. 1 30625 Hannover (Deutschland)

Stephan Ort, Manfred Konrad Structural and functional analysis of deoxyribonucleoside kinases dCK and dGK: design of novel mutant enzymes for therapeutic applications. In mammalian cells, there are four deoxyribonucleoside kinases (dNK) that catalyse the first phosphorylation step in the salvage pathway of nucleosides. These four dNKs have overlapping substrate specificities and differ in their cellular location. Deoxycytidine Kinase (dCK), deoxyguanosine kinase (dGK), and thymidine kinase 2 (TK2) are constitutively expressed throughout the cell cycle and are closely related members of an enzyme family. The fourth dNK, thymidine kinase 1 (TK1), is not sequence-related to the other enzymes and is cell cycle regulated. All dNKs are pharmacologically important because nucleoside analogs are inactive prodrugs that depend on phosphorylation within the cell to gain activity. Human dCK and dGK phosphorylate several clinically important anti-cancer and anti-viral nucleoside analogs such as 1-Β-D-arabinofuranosylcytosine (araC), araG and anti-HIV compounds such as 2',3'-dideoxycytidine (ddC). Sequence comparisons between dCK, dGK and TK2 show a high degree of similarity. One major difference between these 3 dNKs, that might contribute to their different substrate specificities, is an insert of 15 amino acids in dCK and 12 amino acids in dGK that is not present in TK2. To gain insight into how substrate specificities of these dNKs are created we have designed novel mutant dNKs and analysed their enzymatic activities. Promising candidates of these new enzyme variants might be used for therapeutic applications, such as gene therapy approaches. contact: Stephan Ort MPI f. biophysikalische Chemie [email protected] Am Fassberg 11 37077 Göttingen (Deutschland)

Ingo Paarmann, Manfred Konrad Structural requirements for calmodulin binding to membrane-associated guaylate kinases Membrane-associated guanylate kinases (MAGUKs) have been identified at cell-cell contact sites in organisms from Drosophila to man. In mammals, MAGUKs such as SAP90, SAP97, CASK, and p55 constitute a novel superfamily of multidomain proteins that are localized at synapses and participate in the formation of the membrane cytoskeleton. MAGUKs contain at least one PDZ domain, an SH3 domain, a HOOK region, and a domain with striking sequence homology to the known low molecular mass guanylate kinases (GKs). In MAGUK proteins, the SH3 domain interacts intramolecularly with the GK domain. We have analyzed the functional role of the HOOK region localized between the SH3 and the GK domains. Our results clearly show that (Ca2+)4-calmodulin binds to the HOOK regions of both SAP97 and CASK belonging to two different MAGUK subfamilies preferentially under conditions where the intramolecular interaction between the SH3 and the GK domains can occur. Calmodulin binding might stabilize this interaction and serve as an effector molecule to modulate the association of various domain-specific binding partners. contact: Dr. Ingo Paarmann Max-Planck-Institut für biophysikalische Chemie Abteilung für Molekulare Genetik [email protected] Am Fassberg 11, Turm 5 37077 Göttingen (Germany)

Colin Robinson Structure and mechanism of the twin-arginine translocase The twin-arginine translocation (Tat) system operates in the bacterial plasma membrane and the thylakoid membrane of plant chloroplasts. The system recognises proteins bearing N- terminal signal peptides that contain an invariant twin-arginine motif, and has the unique ability to translocate fully-folded proteins across tightly sealed membranes. To date, three important tat genes have been identified in bacteria (tatABC) and homologous genes are present in plants. We have purified a Tat complex from Escherichia coli and hsow that it contains only TatABC, with no evidenmce for the presence of hitherto unidentified proteins. TatB and TatC form the core of the complex and are present in a 1:1 ratio. We further show that a translational fusion between these proteins is fully active, indicating that they form a structural and functional unit within the Tat complex. Preliminary electron microscopy images reveal a stable complex of TatBC that becomes considerably enlarged in the presence of TatA; the architecture of this complex will be discussed in the light of models for the mechanism of this system. The structures and functions of other bacterial Tat systems will also be discussed. contact: Prof. Colin Robinson University of Warwick Department of Biological Sciences [email protected] Coventry (GB)

Oliver Ohlenschläger, Jens Wöhnert, Enrico Bucci, Ramadurai Ramachandran, Roland Zell, Matthias Görlach Structure of an RNA Involved in Enteroviral Replication Enteroviruses are human pathogens responsible for a number of acute and chronic diseases. The initiation of enteroviral positive-strand synthesis requires a ribonucleoprotein complex consisting of a cloverleaf-like RNA structure at the 5´-end of the viral positive-strand RNA, and a set of proteins including the viral protease 3Cpro. The 5´-cloverleaf RNA comprises one stem (A) and three stemloop subdomains (B, C, D). In vitro binding experiments using the 3Cpro of coxsackievirus B3 revealed that the subdomain D of the 5´-cloverleaf is necessary and sufficient for specific 3Cpro binding. Binding analysis using a set of cloverleaf and stemloop D mutants, together with CD spectroscopy, suggested that the apical loop of subdomain D is a major determinant for the specific RNA-protein interactions here (1). To elucidate the structural basis of the recognition of subdomain D by 3Cpro we have determined the structure of a 30-mer RNA from subdomain D by heteronuclear multidimensional NMR spectroscopy. Hydrogen bonds indicating the presence of canonical base pairs and one unusual U:C base pair have been directly detected by the HNN-COSY experiment. Resonance assignments have been obtained using standard triple resonance experiments and a new experiment to link the exchangeable with the H5/H6 protons in pyrimidine bases simultaneously. The stem of stemloop D is in an all-helical conformation and includes a central base paired triple pyrimidine (U:U-U:C-U:U) mismatched region. The apical tetraloop has a defined structure closed by a U:G base pair. Even though the sequence and the closing base pair of the apical tetraloop differ from known stable tetraloops its structure is surprisingly similar to the structure of one of the known stable tetraloops. Literature (1) Zell, Sidigi, Bucci, Stelzner and Görlach, RNA (2002), 8:188-201. contact: Dr. Matthias Görlach Institut für Molekulare Biotechnologie Abt. Mol. Biophys. / NMR-Spektroskopie [email protected] Beutenbergstr. 11 07745 Jena (Deutschland) additional information J. Wöhnert: Institut für Organische Chemie, J.W.G.-Universität, Frankfurt Enrico Bucci: Istituto di Biostrutture e Bioimmagini, Naples, Italy Roland Zell: Institut für Virologie, Friedrich-Schiller-Universität, Jena

Christoph Kluck, Holger Patzelt, Bernd Bukau, Matthias Mayer Structure-function analysis of HscC, the E. coli member of a novel subfamily of specialized Hsp70 chaperones Hsp70 chaperones assist protein folding processes through nucleotide controlled cycles of substrate binding and release. In our effort to understand the structure-function relationship within the Hsp70 family of proteins, we characterized the E. coli member of a novel Hsp70 subfamily, HscC, and identified considerable differences to the well studied E. coli homologue, DnaK, which together suggest that HscC is a specialized chaperone. The basal ATPase cycle of HscC had kcat and Km values that were 8- and 10,000-fold higher than for DnaK. The HscC ATPase was not affected by the nucleotide exchange factor of DnaK GrpE, and stimulated 8-fold by DjlC, a DnaJ protein with a putative transmembrane domain, but not by other DnaJ proteins tested. Substrate binding dynamics and substrate specificity differed significantly between HscC and DnaK. These differences are explicable by distinct structural variations. HscC does not have general chaperone activity as it did not assist refolding of a denatured model substrate. In vivo, HscC failed to complement temperature sensitivity of ΔdnaK cells. Deletion of hscC caused a slow growth phenotype that was suppressed after several generations. Triple knockouts of all E. coli genes encoding Hsp70 proteins (ΔdnaK ΔhscA ΔhscC) were viable, indicating that Hsp70 proteins are not strictly essential for viability. Extensive search for ΔhscC phenotypes revealed a hypersensitivity to Cd2+-ions and UV irradiation, suggesting roles of HscC in the cellular response to these stress treatments. Together our data show that the Hsp70 structure exhibits an astonishing degree of evolutionary plasticity in order to adapt to specialized functions. contact: Dr. Matthias Mayer Universität Heidelberg ZMBH [email protected] Im Neuenheimer Feld 282 69120 Heidelberg (Germany)

Michael Arand Structure-function relationships of epoxide hydrolases Epoxide hydrolases (EH) efficiently detoxify potentially genotoxic epoxides that frequently arise as intermediates in xenobiotic metabolism. In contrast to the many different oxidoreductases that can form these epoxides there are only two separate human EH known to date that detoxify xenobiotic epoxides. Especially one of them, the microsomal EH, combines broad substrate specificity with an apparently high substrate affinity which is mandatory for the proper protection from the hazardous effects of the substrates. The mammalian EH catch and inactivate their substrate in the first step of the enzymatic reaction as a substrate ester intermediate that is subsequently hydrolyzed to form the final product and regenerate the enzyme. The recent X-ray structures of a number of EH help to understand essential details of the reaction. As proposed earlier, epoxide hydrolases possess an α/β hydrolase fold as the central domain. The substrate is hydrolyzed by a catalytic triad in the active site. The major catalytic residue (catalytic nucleophile) is an aspartic acid that forms an ester bond to the substrate molecule under epoxide ring opening. Water activation and ester hydrolysis is carried out by a charge relay system composed of histidin and an acidic residue. A pair of tyrosines that hydrogen bond to the epoxide oxygen during substrate docking supports the nucleophilic attack by proton donation to the oxygen. Kinetic analyses have revealed that the nucleophilic attack is very fast while the subsequent hydrolysis is often much slower, which results in the accumulation of the covalent intermediate and a much lower Km as compared to the "real" dissociation constant and thus a high apparent substrate affinity. Literature 1. Arand M, Grant DF, Beetham JK, Friedberg T, Oesch F and Hammock BD, Sequence similarity of mammalian epoxide hydrolases to the bacterial haloalkane dehalogenase and other related proteins. Implication for the potential catalytic mechanism of enzymatic epoxide hydrolysis. FEBS Lett. 338: 251-256, 1994. 2. Arand M, Wagner H and Oesch F, Asp333, Asp495, and His523 form the catalytic triad of rat soluble epoxide hydrolase. J. Biol. Chem. 271: 4223-4229, 1996. 3. Arand M, Müller F, Mecky A, Hinz W, Urban P, Pompon D, Kellner R and Oesch F, Catalytic triad of microsomal epoxide hydrolase: replacement of Glu404 with Asp leads to a strongly increased turnover rate. Biochem. J. 337: 37-43, 1999. 4. Zou J, Hallberg BM, Bergfors T, Oesch F, Arand M, Mowbray SL and Jones TA, Structure of Aspergillus niger epoxide hydrolase at 1.8 A resolution: implications for the structure and function of the mammalian microsomal class of epoxide hydrolases. Structure Fold. Des. 8: 111-122, 2000. contact: Dr. Michael Arand Universität Mainz Institute of Toxicology [email protected] Obere Zahlbacher Str. 67 55131 Mainz (D)

Christian Lücke, André Richardt*, Heinz Rüterjans, Vicky Katsemi Study on the function and kinetics of diisopropylfluorophosphatase Diisopropylfuorophosphatases (DFPases) are capable of detoxifying chemical warfare agents like diisopropylfluorophosphate (DFP), soman, sarin, tabun and cyclosarin by hydrolysis. These organophosphorus triesters act as nerve agents by irreversibly inhibiting the acetylcholinesterase [1]. The DFPase reported here was originally isolated from head ganglion of Loligo vulgaris, but the protein used in this work has been recombinantly expressed in E. coli. It hydrolyzes DFP three to four times faster than any of the other nerve agents [2]. This ‘squid-type’ DFPase contains a high- and a low-affinity calcium binding site. The high-affinity calcium is essential for the structural integrity of the protein [3], whereas the low-affinity calcium has been shown to play a crucial role in the catalysis reaction [3, 4]. Based on the crystal structure, a reaction mechanism has been proposed [4]. Since neither the biological function of the enzyme is known, nor an inhibitor could be found, the proposed mechanism has been further explored by means of site-directed mutagenesis as well as by kinetic studies. In this work several residues that are important for DFP hydrolysis have been identified as well as their effects on the hydrolysis of sarin, soman and tabun determined. Moreover we were able to demonstrate the relative importance of the hydrophobicity of the binding pocket and of the charges in the neighborhood of the Ca2+ atoms. Literature 1. F.C.G. Hoskin; Science, 172 1243-1245 (1971) 2. W. Schulz, B. Schäfer, G. Stroop, and H. Rüterjans; “Forschungsbericht aus der Wehrtechnik,” Auftragsnummer T/R 770/D 00001/D 1750 (1987) 3. J. Hartleib, S. Geschwindner, E.I. Scharff and H. Rüterjans; Biochem. J., 353, 579-589 (2001) 4. E.I. Scharff, J. Koepke, G. Fritzsch, C. Lücke and H. Rüterjans; Structure, 9, 493-502 (2001) contact: Dipl.-Biologin Vicky Katsemi J. W. Goethe Universität Biophysikalische Chemie [email protected] N230, 1OG, Marie-Curie-Str. 9 60439 Frankfurt am Main (Deutschland) additional information *German Armed Forces Scientific Institute for Protection Technologies (WIS), Munster, Germany

Cordula Enenkel Targeting mechanism of proteasomes to nuclear / ER membranes and into the nucleus of yeast Andrea Lehmann, Marion Fehlker, Petra Wendler, Ulf Dühring, Peter-Michael Kloetzel and Cordula Enenkel* Institut für Biochemie, Humboldt-Universität / Charité, Monbijoustr. 2, 10117 Berlin. 20S proteasomes are matured from short-lived precursor complexes. In our recent work we report on Ump1-associated precursor complexes to be predominantly present in the nuclear / ER fraction of yeast. Our data suggest that precursor complexes are imported into the nucleus via the classical NLS receptor before being finally matured into 20S proteasomes (1). To elucidate Ump1-independent proteasome maturation pathways, we analyzed the subcellular distribution of precursor complexes in ump1 deletion mutants by following proproteins especially of those beta subunits that are incorporated late into precursor complexes. Pro-beta5 subunits, whose processing is vital, are found in high molecular mass complexes and appear to be mainly localized inside the nucleus of ump1 deletion mutants. We are currently investigating whether Ump1-lacking precursor complexes are imported into the nucleus via the cNLS receptor. According to our working hypothesis, both soluble receptors that target proteasomal subcomplexes to NE / ER membranes and membrane-anchored receptors for proteasomal subcomplexes may exist. We identified a high molecular mass protein within proteasomal complexes of the nuclear / ER fraction. By native gel electrophoresis this new protein migrates with a not-yet characterized form of the 20S proteasome and an Ump1-containing precursor complex. Our data suggest that this new protein is involved in the final assembly of 26S proteasomes. Literature (1) Lehmann, A., Janek, K., Braun, B., Kloetzel, P.-M. and Enenkel, C. (2002) J. Mol. Biol. 317, 401-413. contact: Dr.rer.nat. Cordula Enenkel Humboldt Universität / Charité Institut für Biochemie [email protected] Monbijoustr. 2 10117 Berlin (Germany)

Joao Pedro Marques, Ingrid Dudeck, Martin Schattat, Ralf Bernd Klösgen Targeting of chimaeric GFP-polypeptide fusions within chloroplasts In order to get further insights into the transport specificity of the different thylakoid import pathways, we have performed in vitro import experiments using protein fusions among the GFP protein and several thylakoid transit peptides. The later were part of proteins assigned to both the delta pH/TAT (16 and 23 kDa subunits of the water splicing complex of PS2) and Sec pathways (plastocyanin and 33 kDa subunit of the water splicing complex of PS2). In the presence of isolated pea thylakoids we were able to demonstrate the efficient and specific targeting of 16/GFP and 23/GFP fusion proteins by the delta pH pathway to the thylakoid lumen. In contrast, even after the addition of isolated stroma fraction, no lumenal import of the PC/GFP and 33/GFP could be observed in isolated pea thylakoids. In the presence of intact pea and spinach chloroplasts, however, besides an efficient import of the 16/GFP and 23/GFP fusions, a low level of 33/GFP import into the thylakoid lumen could also be noticed. The data obtained suggest a preferential import of GFP fusions into the thylakoid lumen via the delta pH pathway. The explanation for this behaviour might reside in the transport specificity of the delta pH pathway for folded proteins. In order to assay the folding status of GFP during the course of plastid import and internal sorting, transgenic Arabidopsis lines expressing GFP fusions are currently being produced. contact: Doktor Joao Pedro Marques MLU-Halle Institut für Pflanzenphysiologie [email protected] Weinbergweg 10 06120 Halle (Saale) (Germany)

Heinrich Jung, Maike Rochon, Corinna Weber-Sparenberg Targeting of proteins to the flagellar type III secretion system of Escherichia coli A number of Gram-negative animal and plant pathogenic bacteria use type III secretion systems to inject bacterial effector proteins into eukaryotic cells. The type III secretory pathway is also required for transport of flagellar structural proteins beyond the cytoplasmic membrane whereby playing an essential role in the biogenesis of the bacterial flagella. The molecular mechanism underlying this process is still enigmatic. Studies on type III systems involved in bacterial virulence suggest that protein substrates are marked for secretion via a mRNA mediated signal and by their cognate chaperones. Alternatively substrates may be recognized by a N-terminal amino acid sequence 1. To investigate substrate recognition during flagella biogenesis, we used FlgD, a scaffolding protein in flagellar hook assembly, as a model substrate. Type III system-dependent export of a FlgD-PhoA hybrid into the periplasm of E. coli was determined enzymatically as well as by Western blotting. Truncation-analysis of FlgD reveals that the N-terminal 83 amino acids are crucial for export. Furthermore, frame shift mutations altering the sequence of amino acids 2 to 20 of FlgD prevent export of FlgD-PhoA, whereas replacement of N-terminal amino acids with an amphipathic sequence stimulates the transport of the hybrid protein. Further studies showed that the amphipathic sequence is essential but not sufficient for FlgD-PhoA transport. Literature 1 S.A. Lloyd, A. Forsberg, H. Wolf-Watz, M.S. Francis (2001) Trends Microbiol. 9, 367-71. contact: Corinna Weber-Sparenberg Universität Osnabrück Mikrobiologie [email protected] Barbarastr. 11 49069 Osnabrück (Germany) additional information PD Dr. Heinrich Jung Universität Osnabrück Mikrobiologie Barbarastrasse 11 49069 Osnabrück Germany [email protected]

Peter Heimann, Harald Jockusch, Christiane Wiegand Temperature-sensitive TMV Coat Proteins as Pathogens in Animal Cells The rod shaped particles of tobacco mosaic virus (TMV) consist of a 6Kb single stranded RNA and about 3000 identical subunits of the 158 amino-acid-residue coat protein (CP). Wildtype (temperature resistant, tr) TMV forms paracrystalline masses in host plant cells but the infected tissue survives and grows. Single amino acid replacements in TMV-CP may, at moderately elevated temperature, destroy its ability to form rods and cause dramatic cytopathic effects, thus acting as toxic structural proteins [1]. We have used TMV-CP tr wildtype and temperature-sensitive (ts) mutant constructs to transfect animal cells to test whether the ts mutants are toxic also in a heterologous system. Cell lines CHO, COS-7 and NSC (neuronal) were used. At 37° C, tr TMV-CP was diffusely distributed in the cytoplasm whereas with mutant ts TMV-CPs distinct lumps of antigen were observed, presumably consisting of denatured aggregated CP. COS-7 cells containing these lumps were inflated and vacuolized, and thus pathologically altered in contrast to tr CP transfected cells. Several neurodegenerative disease are thought to be caused by mutant structural proteins with altered conformations, solubility or degradation characteristics. TMV-CP and its mutants in mammalian cells, provide a "synthetic" model for toxic protein deposits which can be manipulated at will and studied with respect to stress responses (as already shown in plant cells, e.g. HSPs [2], ubiquitination [3]). Supported by Fonds der Chemischen Industrie. Literature 1. Jockusch, H. (1968) Naturwiss. 55, 514-518 2. Jockusch, H. et al. (2001) Mol. Plant-Microbe Interact. 14, 914-917 3. Jockusch, H., and Wiegand, C. in preparation contact: Dipl.Biol. Christiane Wiegand Uni-Bielefeld Entwicklungsbiologie & Molekulare Pathologie [email protected] Universitätsstr. 25 33516 Bielefeld (Deutschland)

Volkmar Gieselmann, Uwe Rauch, Karl Schilling, Penka Pesheva, Sebastian Franken, Stephan L. Baader, Joachim Kappler Tenascins are associated with lipid rafts isolated from mouse brain Lipid rafts are microdomains of the plasma membrane which are enriched in glycosphingolipids and specific proteins. The reported interactions of several raft-associated proteins (such as e.g. F3) with tenascin C and tenascin R prompted us to consider that these oligomeric multidomain glycoproteins of the extracellular matrix (ECM) could associate with rafts. Here, we show punctate immunocytochemical distributions of tenascin C (TN-C) and tenascin R (TN-R) at the membrane surface of neural cells resembling the pattern reported for raft-associated proteins. Moreover, cholesterol depletion with methyl-b-cyclodextrin reduced the punctate surface staining of TN-C. Consistently, TN-C was associated with lipid rafts of neonatal mouse brain according to sucrose density gradient centrifugation experiments. Furthermore, TN-R, was also found in rafts prepared from myelin of adult mice. Thus, brain-derived tenascins are able to associate with lipid rafts. contact: Dr. Joachim Kappler Universität Bonn Physiologische Chemie [email protected] Nussallee 11 53115 Bonn (Deutschland)

heiko funke-kaiser, julia lemmer, christian von langsdorff, alexander thomas, slobodan d. Kovacevic, meike strasdat, frank zollmann, martin paul, hans-dieter orzechowski The cardiac transcription factor Nkx2-5 regulates the alternative promoters of the endothelin-converting enzyme-1 gene in H9c2 cardiomyoblasts Mouse knock-out models have established a fundamental role for the homeobox transcription factor Nkx2-5 and for the metalloprotease, endothelin-converting enzyme-1 (ECE-1), in heart development. Here, we demonstrate for the first time a functional link between Nkx2-5 and ECE-1. Using luciferase reporter gene assays, site-directed mutagenesis and gel shift assays we show that Nkx2-5 regulates the alternative promoters of ECE-1 isoforms. In transiently transfected rat H9c2 cardiomyoblasts, the promoters specific for ECE-1a, ECE-1b and ECE-1c are activated by Nkx2-5 coexpression. Functional analysis of ECE-1 promoter constructs, harbouring point mutations in Nkx2-5 consensus sequences, and gel shift assays support an indirect mechanism of activation in case of ECE-1a and -1c promoters, whereas the ECE-1b promoter might also be regulated directly. In addition, the activating effect of Nkx2-5 on ECE-1a, ECE-1b and ECE-1c mRNA expression was also confirmed in the chromatin context by northern blot or real-time PCR analysis respectively, using stably transfected cells overexpressing Nkx2-5. The interaction presented in this work provides a possible explanation of certain phenotypic aspects of patients with Nkx2-5 mutations and may also be of significance for the pathophysiology of heart failure.

contact: dr heiko funke-kaiser freie universität berlin, UKBF klinische pharmakologie und toxikologie [email protected] hindenburgdamm 30 12200 berlin (deutschland)

Gabi Bauer-Manz, Andreas Kuhn, Lambertus van den Berg, Justyna Serek The E.coli membrane insertase YidC: A Chaperone for Protein Folding into the Membrane YidC, the homologe of the mitochondrial inner membrane protein Oxa-1 has been cloned from the E.coli chromosome, modified with a his-tag and purified to homogeneity. The 60kD protein was reconstituted with E. coli phospholipids for proteoliposomes. These proteoliposomes were capable in supporting the insertion of purified membrane proteins in chemical amounts. We suggest that YidC is a membrane insertase that promotes membrane protein insertion. The two model proteins we tested for membrane insertion in vitro, were the single-spanning Pf3 coat protein and the double-spanning M13 procoat protein. Both have been recently shown to depend on YidC for membrane insertion in vivo (1,2). The Pf3 coat protein was purified by reverse phase and size exclusion chromatography and soluble in 100mM Tris-HCl pH 8,5, 10% isopropanol. The M13 procoat protein was purified by affinity chromatography. When the proteins were added to the YidC containing proteoliposomes the proteins readily integrated in a transmembrane configuration. This was tested by addition of protease to the outside showing that the integrated proteins were protected from proteolysis. In the absence of YidC both proteins bind to the membrane surface but only minute amounts are resistant to the protease and therefore translocated across the membrane. Since the YidC protein catalytically supports the integration of the two model proteins we propose that it acts similar to a chaperone by binding the non-translocated form of the substrate and releasing the translocated membrane protein. Literature 1.Samuelson, J.C., F. Jiang, L. Yi, M.Chen, A.Kuhn and R.E.Dalbey (2001). Funktion of YidC for the insertion of M13 procoat protein in E. coli:Translocation of mutants that show differences in their membrane potential dependence and Sec-requirement. J. Biol Chem.276,34847-34852. 2.Chen, M., J. samuelson, F.Jiang, M. Müller, A.Kuhn and R.Dalbey (2002) Direct interaction of YidC with the Sec-independent Pf3 coat protein during ist membrane insertion. J.Biol.Chem. 277, 7670-7675. contact: Justyna Serek Universität Hohenheim Mikrobiologie und Molekularbiologie [email protected] Garbenstr.30 70599 Stuttgart (Baden Württemberg)

Leopold Flohé The element of the moon moonlighting for fertility Selenium is contained in spermatozoa primarily as an integral part of phospholipid hydroperoxide glutathione peroxidase (GPx-4). In testis, GPx-4 is expressed as a cytosolic, a mitochondrial and a nuclear variant, most abundantly in spermatids. The differentiation of spermatids to mature spermatozoa is accompanied by a shift of the redox equilibrium characterized by a loss of GSH and oxidation of protein SH groups. In parallel, GPx-4 is transformed from an active peroxidase into an enzymatically inactive structural protein that represents the major component of the mitochondrial capsule.1 Biochemically, this transformation of GPx-4 is explained by the use of protein SH groups as alternate substrates, which results in protein corss-linking via Se-S bonds. The clinical relevance of this “moonlighting” process for male fertility is underscored by significant correlations between GPx-4 content of sperm and fertility-related parameters such as sperm motility and morphological integrity.2 Further, clustered single base polymorphisms were detected in gpx-4 of patients with idiopathic infertility.3 It is concluded that selenium deficiency may compromise male fertility by impairing GPx-4 synthesis, yet infertility may equally be due to genetic gpx-4 variants or any other disturbance of the time- and site-specific gpx-4 expression or the GPx-4 transformation process in late spermatogenesis. Literature 1F. Ursini, S. Heim, M. Kiess, M. Maiorino, A. Roveri, J. Wissing, L. Flohé, Science 285, 1393 (1999) ; 2C. Foresta, L. Flohé, A. Garolla, A. Roveri, F. Ursini, M. Maiorino Biol. Reprod. (2002), in press 3M. Maiorino, V. Bosello, F. Ursini, C. Foresta, M. Scapin, L. Flohé Biol. Reprod. submitted contact: Prof. Dr. Leopold Flohé Universität Braunschweig Institut für Biochemie [email protected] Mascheroder Weg 1 38124 Braunschweig (Deutschland) additional information L. Flohé,* M. Maiorino,** S. Pilawa,* H. Stzajer,* and F. Ursini,** * Dept. Biochemistry, Technical University of Braunschweig, D-38124 Braunschweig; ** Dept. Biological Chemistry, University of Padova, I-35121 Padova

Heide Schmid, Thai-Hoa Nguyen-Xuan, Minh-Huy Tran, Gerold Schwarz, Hubert Kalbacher, Heinrich Wiesinger The endosomal/lysosomal distribution of cathepsins (cat) B, L and S and N-acetyl-ß-D-glucosaminidase (NAG) is differentially modulated in murine cell lines of microglial and monocytic origin Lysosomal hydrolases are involved in turnover of cellular constituents and therefore highly regulated by metabolic conditions. Moreover, in immunocompetent cells like phagocytes endosomal cathepsins act as antigen-processing enzymes. In order to assess such functional differences in murine microglial (N11) and monocytic (RAW 264.7) cell lines, the catalytic activity of the lysosomal marker enzyme NAG was compared with that of cathepsins B, L and S in crude organelle preparations, endosomal and lysosomal fractions under different culture conditions. In contrast to RAW 264.7 cells N11 cells exhibited higher endosomal than lysosomal activities of all four enzymes when cultured in DMEM with 10 % FCS. 24 h of culture in DMEM with 1 % FCS and 1 mM L-NAME resulted in reduced organellar enzyme activities of both cell lines with the exception of cat L and cat S in N11 cells, however, with lysosomal enzyme activities exceeding endosomal activities. Simulation of immunomodulatory conditions with interferon-γ (200 U/ml) reduced the ratio of endosomal versus lysosomal enzyme activities in N11 cells, but increased this ratio in RAW 264.7 cells. Thus this cytokine may convert N11 cells into a more immunogenic state, RAW 264.7 cells, however, into a more degradative state. contact: Dr. rer.nat. Heide Schmid Universitätsklinikum Institut für Pathologie [email protected] Liebermeisterstr. 8 72076 Tübingen (Deutschland) additional information H. Schmid, T.-H. Nguyen-Xuan, M.-H. Tran, Institut für Pathologie, Liebermeisterstr. 8, G. Schwarz, H. Kalbacher, H. Wiesinger, Physiologisch-chemisches Institut, Hoppe-Seyler-Str. 4, Universität Tübingen, D-72076 Tübingen

Astrid Schön, Olaf Gimple, Christian Heubeck The evolution of RNase P in eukaryotic organelles Many essential metabolic reactions are dependent on nucleic acids as cofactors or catalysts. Examples are reactions in nucleic acid biosynthesis, RNA-dependent conversions of small metabolites, and RNA processing reactions. Many of of these pathways are highly conserved in extant organisms, suggesting an ancient origin in an "RNA world". The essential tRNA processing enzyme RNase P is a striking example for enzyme evolution: it is a ribonucleoprotein (RNP) in bacteria, archea, and eukaryotic nuclei, with an essential RNA component. This RNA is a true ribozyme in bacteria and some archaea. In contrast, no RNA component exists in the mitochondria and chloroplasts of multicellular organisms, posing the question of enzyme evolution in these bacteria-derived organelles. The primitive plastids from certain unicellular algae, however, contain an RNP-type RNase P. Their essential RNA components are similar to those of cyanobacteria, and show high structural diversity. The overall composition of the plastid RNase P, with a rather large protein contribution, is more similar to the eukaryote- than to the bacterial-type enzymes. However, as in nuclear RNase P, the RNA is an essential enzyme component, but is not catalytically active as a ribozyme. Surprisingly, a fully active RNase P holoenzyme can be reconstituted from the corresponding organelle RNAs and cyanobacterial protein subunits. This suggests that, despite the observed structural diversity, the catalytic activity of the enzyme still resides in the RNA subunit.

Literature Baum, M., Cordier, A. and Schön, A. (1996). J. Mol. Biol. 257, 43-52. Cordier, A. and Schön, A. (1999). J. Mol. Biol. 289, 9-20. Heubeck, C. and Schön, A. (2001). In: Meth. Enzymol. 342, 118-134. Gimple, O. and Schön, A. (2001). Biol. Chem. 382, 1421-1429. contact: Dr. Astrid Schön University of Leipzig Biochemistry [email protected] Talstr.33 04109 Leipzig (Germany) additional information Adress of Co-Authors: Institute of Biochemistry, Biozentrum, Am Hubland, 97074 Würzburg

Detlef PIETROWSKI, Gia Anh Bao PHAN, Paulina WITT, Christoph KECK THE EXPRESSION OF ANGIOGENIC GROWTH FACTORS IN HUMAN GRANULOSA CELLS ARE CONTROLLED BY HUMAN CHORION GONADOTROPIN. Introduction: Human granulosa cells (GC) derived from IVF-procedures are a well established model system to study the regulation of various cytokines and growth factors involved in corpus luteum development and function in vitro. Human chorion gonadotropin (hCG) is required for growth and development of the corpus luteum from avascular GCs in vivo. To nourish the growing corpus luteum a dramatic vessel growth into this developing gland take place. Here we characterise and analyse the influence of hCG on the expression of various growth factors involved in angiogenesis in human GC. Material and methods: GCs were isolated from follicular fluid of patients undergoing IVF therapy and cultured as described elsewhere (Keck et al., 1998). After 48 hours incubation in Medium 199 / 5% FCS cells were stimulated with hCG (10U/ml). RNA was isolated by the method of Chomczynski and Sacchi (1987) and messenger RNA expression of VEGF-A and related molecules (VEGF-B, VEGF-C, VEGF-D, PGF, PDGF-A, PDGF-B) as well as messenger RNA of Angiopoietins (ANG), ephrins, tissue inhibitor of matrix metalloproteinases 1 (TIMP-1), meningioma associated complimentary DNA (mac25) and connective tissue growth factor (CTGF) was determined by RT-PCR. Results were relatively quantitated by TaqMan Real Time PCR or semiquantitated by densitometry and the amount of protein were determined by ELISA techniques. Results: We could demonstrate that the expression level of angiogenic factors VEGF-A, ANG-2, TIMP-1 and CTGF significantly changed during the stimulation of GC with hCG. Whereas VEGF-A and TIMP-1 expression increases during the incubation time, the expression of ANG-2 and CTGF is remarkable decreased. Additionally, we provide evidence that the regulation of Ang-2 and VEGF-A in GC is differentially mediated via the PKA and PKC dependent signalling cascades. Conclusions: Human chorion gonadotropin has a significant influence on the expression of growth factors involved in angiogenesis in the human corpus luteum. It mediates both the upregulation and the downregulation of factors which are involved in blood vessel proliferation as well as blood vessel maturation and stability. We hypothesize that hCG influences the growth of the corpus luteum by mediating the angiogenic process in this developing gland. contact: Dr.med. Gia Anh Bao PHAN Universität Freiburg Universitäts-Frauenklinik [email protected] Hugstetterstrasse 79106 Freiburg im Breisgau (Deutschland)

Christina Deschermeier, Georg Mayr, Werner Fanick, Marcus Nalaskowski The human inositol phosphate multikinase (IPMK) is targeted to the nucleus of mammalian cells by a basic cluster in its C-terminus Four proteins (ArgRI, ArgRII, ArgRIII, Mcm1) regulate the expression of several genes involved in the metabolism of arginine in yeast. In the presence of arginine, they repress the synthesis of five anabolic enzymes and induce the synthesis of two catabolic enzymes. It was previously reported that an ARGRIII deletion yeast mutant shows dramatic changes in its inositol phosphate levels, indicating that its multikinase activity is in-vivo important for the maintenance of the balance of the different inositol phosphates. Deletion yeast mutants also show an accumulation of poly(A) RNA in the nucleus caused by an impediment to nuclear mRNA export. We present here the cloning of a full-length 1248 bp cDNA encoding a human inositol phosphate multikinase (IPMK). This protein has a calculated molecular mass of 47.219 kDa. Functionally important motifs are conserved between the human IPMK and yeast ArgRIII. NRK 52E cells were transiently transfected with the HsIPMK cDNA fused with a C-terminal and a N-terminal fluorescent protein tag, respectively. Both fusion proteins were localized predominantly within the nuclei of the transfected cells. By sequence analysis we found that the position of the basic residues in a C-terminal cluster is conserved between this sequence and a fragment from the human c-myc protein, for which a weak NLS activity was shown. It can be concluded from our results that this basic cluster is positively involved in the nuclear targeting, because its functional elimination changes the predominantly nuclear localization to an equal distribution between nucleus and cytoplasm. These findings are consistent with the concept of a nuclear inositol phosphate signalling and phosphorylation pathway in mammalian cells. Literature Dubois, Dewaste, Erneux and Messenguy (2000) FEBS Lett. 486, 300-304 El Bakkoury, Dubois and Messenguy (2000) Mol. Microbiol. 35, 15-31 Odom, Stahlberg, Wente and York (2000) Science 287, 2026-2029 Saiardi, Nagata, Luo, Sawa, Luo, Snowman and Snyder (2001) Proc. Natl. Acad. Sci. U S A 98, 2306-2311 Shears (2000) Bioessays 22, 786-789 contact: Marcus Nalaskowski Universität Hamburg Institut für Medizinische Biochemie [email protected] Martinistraße 52 20246 Hamburg (Deutschland)

Armin Huber, Reinhard Paulsen, Claudia Franz The INAD signaling complex of Drosophila photoreceptor: Recombinant Expression and Assembly in a Cell Culture System In fly photoreceptors phototransduction is accomplished by a prototypical G-protein coupled signaling pathway which is in part assembled into a multimolecular signaling complex. The core components of the signaling complex, comprising a phospholipase C (PLC²), an eye-specific protein kinase C (ePKC) and the major light-activated cation channel TRP, are tethered to the PDZ domains of INAD which constitutes a scaffold for the complex (Huber et al., 1996; Chevesich et al., 1997; Tsunoda et al., 1997). The correct localization to the rhabdomeral photoreceptor membrane and the stability of these core proteins depends on the proper assembly of the signaling complex. The site and the mechanism of the assembly process are not yet investigated in detail. We have examined the suitability of Drosophila S2 cells, which have previously been used to express TRP and TRPL channels (Hardie et al., 1997), for recombinant expression of the INAD signaling complex of Drosophila melanogaster. We succeeded to express the four major complex components TRP, ePKC, PLC² and INAD in S2 cells. The recombinant expression of the signaling proteins was verified immunocytochemically as well as via Western Blot analysis. We carried out co-immunoprecipitation studies to investigate wether or not the INAD core signaling complex is assembled in the cell culture system. So far, we showed the co-immunoprecipitation of all four INAD signaling complex components recombinantly expressed in S2 cells. Literature Huber et al. (1996) EMBO J. 15: 7036-7045 Chevesich et al. (1997) Neuron 18: 95-105 Tsunoda et al., 1997 Nature 388: 243-249 Hardie et al., 1997 Cell Calcium 21: 431-440 contact: Claudia Franz Universität Karlsruhe Zell- und Neurobiologie [email protected] Haid-und-Neu-Str. 9 76131 Karlsruhe (Germany)

Anne Navarrete Santos, Sarah Tonack, Michaela Kirstein, Bernd Fischer The insulin receptor and the insulin-dependent glucose transporter isoform 4 are expressed in rabbit preimplantation embryos Glucose is the most important energy substrate for mammalian blastocysts. Glucose uptake is mediated by glucose transporters. In muscle and adipocyte cells insulin stimulates glucose uptake by activation of the insulin receptor (IR) pathway and translocation of glucose transporter 4 (GLUT4) from the cytoplasm to the outer cell membrane. In mammalian embryos an insulin treatment during embryo culture has beneficial effects for embryo development with an increase in cell number, protein and DNA synthesis. The mechanism of insulin action in embryonic cells, however, is still not clear. So far, insulin stimulated glucose uptake and expression of GLUT4 has not been shown in embryos. Investigating bovine embryos, we were able to demonstrate for the first time the expression of GLUT4 in mammalian preimplantation embryos in a stage-specific manner [1,2]. In present study the expression of the IR and Glut4 was studied in rabbit blastocysts using RT-PCR, in situ hybridisation, western blot and immunohistochemistry. The rabbit mRNA sequences for the 3690bp coding region of IR and the 3'-(1600bp) terminus of rabbit Glut4 were determined by RACE-PCR and sequencing. Using specific primers for RT-PCR, we found GLUT4 being expressed in 3 days old morulae and in 4 and 6 days old blastocyst. The IR transcript was detectable only in the blastocyst stage. The protein expression of IR and GLUT4 in blastocysts was confirmed by western blotting. GLUT4 is localised in both the embryoblast and trophoblast cells. Our results on IR and GLUT4 expression in rabbit blastocysts could explain the positive effects of insulin in early embryonic development and indicate (i) a developmental control and (ii) an insulin-sensitive glucose transport in mammalian blastocysts. Literature [1] Navarrete Santos A, Augustin R, Lazzari G, Galli C, Sreenan JM, Fischer B (2000) The insulin-dependent glucose transporter isoform 4 is expressed in bovine blastocysts. Biochem. Biophys. Res. Comm. 271, 753-760 [2] Augustin R, Pocar P, Navarrete-Santos A, Wrenzycki C, Gandolfi F, Niemann H, Fischer B (2001) Glucose transport is developmentally regulated in in vitro derived bovine preimplantation embryos. Mol. Reprod. Dev. 60, 370-6 contact: Dr. Anne Navarrete Santos MLU Inst. f. Anatomie u. Zellbiologie [email protected] Gr. Steinstraße 52 06097 Halle (BRD)

Jens Wulfaenger, Alexander Navarrete Santos, Tanja Blosz, Juergen Langner, Dagmar Riemann The intracellular N-terminus of aminopeptidase N/CD13 affects association with caveolae Aminopeptidase N (APN)/CD13 is a zinc-dependent metalloprotease with ubiquitous expression and a broad functional repertoire. It is a type II-integral membrane protein with a short (8-10 amino acids) cytoplasmic domain. APN modulates signal transduction by processing bioactive peptides (enkephalins, kinins, chemokines). Otherwise, APN ligation via antibodies is sufficient to activate intracellular signalling in monocytes (increase in [Ca2+]i, MAP kinase activation). A possible explanation could be the association of APN in specialized membrane microdomains, also referred to as rafts or caveolae, which are involved in signal transduction. To investigate the role of the intracellular part in the signalling via APN, a mutant lacking the intracellular tail was constructed and transiently transfected in CHO-K1 cells. Mutation did not change neither membrane localization nor enzyme activity of APN. However, raft association was strongly diminished in the tailless variant. Further studies have to deal with the function of the intracellular part in signal transduction. contact: Dr. med. Dagmar Riemann Martin-Luther-University Halle/Wittenberg Institute of Med. Immunology [email protected] Magdeburger Str. 2 06097 Halle (Germany)

Jeroen Buters The knockout approach to understand the function of xenobiotic metabolising enzymes Toxicology is a science that makes predictions of the toxicity of compounds in humans. Extrapolating results from animals to the human situation is hampered by many drawbacks. Physiological modeling is the method of choice. However, to make a good physiological model detailed knowledge of the mechanism of toxicity is needed. Mechanisms in toxicology often include a balance between competing metabolic pathways, some leading to detoxification and some leading to toxic effects. Important for modeling is the knowledge of the toxicity determining steps in vivo. Cytochrome P450 are often the rate limiting step in the metabolic activation of compounds to toxic or carcinogenic metabolites. Because cytochrome P450 is a superfamily of 57 human enzymes with overlapping substrate specificity, cytochrome P450 knockout mice proved to be excellent models to determine the most important metabolic step in vivo. For instance cytochrome P450 1A1, CYP1A1, was for decades thought to be the main activating enzyme of polycyclic aromatic hydrocarbons (PAH) to carcinogenic metabolites. PAH are present in food, diesel exhaust gases, cigarette smoke etc.. Knockout mice for CYP1A1 however developed more, not less tumors after PAH exposure. In contrast, CYP1B1, previously an unknown cytochrome P450, knockout mice were resistant to PAH induced tumors. It is thus important to look at the polymorphisms of CYP1B1, and not CYP1A1, for the etiology of i.e. PAH induced lung cancers. These toxic mechanism are obtained in mice with mice cytochrome P450. To bridge the gap to the human situation, the critical enzyme of mice and it homologue in humans can be cDNA-expressed and compared. More interesting are humanized mice, knockout for the mouse enzyme and transgenic for the human enzyme, including human regulatory elements (up to 10 Mbases constructs). Conditional knockouts are not very useful for cytochrome P450 as all cytochrome-null mice born to date were viable and without any obvious defects. Deleting cytochrome P450 oxidoreductase, the energy donating enzyme to CYP, proved lethal and conditional knockouts of this enzyme have since been made. This shed light on the physiological role of cytochrome P450. Mice, and perhaps man can do without one or two cytochromes P450, but die when they lack all functional cytochromes. contact: PD Dr. Jeroen Buters Zentrum f. Allergie und Umwelt [email protected]> Biederstreinerstr. 29 80802 München (D)

Ralf Bernd Klösgen, Bo Hou The mechanism of deltapH/TAT-dependent protein translocation across the thylakoid membrane Protein translocation into or across the thylakoid membrane is accomplished by at least four distinct pathways: the Sec-dependent pathway, the SRP-dependent pathway, the pH/TAT-dependent pathway and the spontaneous protein insertion pathway. The pH/TAT-dependent protein translocation pathway is the sole system that is capable of transporting both folded and unfolded proteins across the thylakoid membrane. Translocation of precursors by this pathway does not need stromal factors or nucleoside triphosphates, but is strictly dependent on the thylakoidal pH. In order to analyze the mechanism of pH/TAT-dependent protein translocation, we have constructed a chimeric 16/23 protein, which consists of the transit peptide of the 16 kDa protein and the mature part of the 23 kDa protein. Like the two corresponding authentic precursor proteins (pre-16 kDa and pre-23 kDa, respectively), the chimeric 16/23 is targeted by pH/TAT-dependent pathway. During thylakoid import, the chimeric protein is retarded in the pH/TAT-dependent translocation machinery, resulting in transmembrane translocation intermediates. Two distinct steps of the transport process have been identified, which are characterized by different topologies of the translocation intermediates. Taking advantage of this finding, we could make detailed studies on the mechanism of each processing step. Data on the differences between these two steps, e.g. considering their dependence upon pH/TAT-translocase or the energy requirement, will be presented. contact: Master Bo Hou Martin-Luther-Universität Halle-Wittenberg Institut für Pflazenphysiologie [email protected] Weibergweg 10 06120 Halle (Saale) (Germany)

Hassan Dihazi, Klaus Eschrich The Mechanism of regulation of yeast 6-phosphofructo-2-kinase by glucose and hypertonic stress We investigated the posttranslational regulation of the metabolic signaling enzyme PFK2 in Saccharomyces cerevisiae. Both glucose and hypertonic stress induce the in vivo phosphorylation and concomitant activation of yeast PFK2. The activation of PFK2 by glucose was chieved by a single phosphorylation near the COOH-terminus of the protein (Ser644) and an additional fivefold phosphorylation near the NH2-terminus on the peptide fragment T67-101. The Ser644 site was confirmed by in vitro phosphorylation with protein kinase A. Increasing external osmolarity causes a threefold activation of PFK2 accompanied by the multiple phosphorylation of the protein of the same NH2-terminal peptide fragment as by glucose induction. The nature of the responsible protein kinase(s) is unknown. Furthermore, we have shown the main role of PFK2 in the maintenance of the internal osmolarity. The activation of PFK2 and the resulting increase of the rate of glycolysis are essential for survival and growth of yeast cells in hypertonic environment. Yeast cells expressing PFK2 accumulate three times more glycerol than cells without PFK2 which are not able to grow in the presence of high salt concentrations. contact: Hassan Dihazi University Leipzig Biochemistry [email protected] Liebig strasse 16 04103 Leipzig (Germany) additional information Eschrich, K., Institute of Biochemistry, Medical Faculty, University of Leipzig, Liebigstrasse 16, Leipzig 04103, [email protected]

Brigitte Schmitz, Julia Kellersmann The O-GlcNAc modification of proteins interferes with signaling by PKA and CDK5 dependent phosphorylation in neurons N-acetylglucosamine linked O-glycosidically (O-GlcNAc) to serine and threonine residues of proteins is a posttranslational modification of proteins of the cytosol, the nucleus and of the cytosolic domains of transmembrane proteins (Wells et al. 2001). We have previosly shownthat the O-GlcNAc levels of proteins from neurons in culture respond reciprocally to inhibition or activation of phosphorylation by treatment of cells with specific pharmacological agents. Vice versa, inhibiton of O-GlcNAc hydrolase with the specific inhibitor PUGNAc results in increased O-GlcNAc levels of proteins. Extending these earlier studies we show that treatment of N2a neuroblastoma cells with PUGNAc and with the PKA activator dbcAMP paradoxically leads to an increase in the O-GlcNAc level of several proteins which is significantly higher than in the presence of PUGNAc alone. Addition of inhibitors in the reverse order did not yield increased O-GlcNAc levels. Since CDK5 is a downstream target of PKA in neurons, cells were treated with PUGNAc and the CDK5 inhibitor roscovitine. The same strong increase in O-GlcNAc expression was detected as in the presence of PUGNAc and dbcAMP. Western blot studies of immunprecipitates indicate that CDK5 is O-GlcNAc-modified, suggesting that in addition to phosphorylation the O-GlcNAc modification may regulate the activity of CDK5. These observations provide further evidence that signaling mechanisms are modulated by a cross talk between O-GlcNAcylation and phosphorylation. Literature Wells et al.,Science 291, 2376-2378 (2001) contact: Julia Kellersmann Uni Bonn Institut für Physiologie, Biochemie und Hygiene der Tiere [email protected] Katzenburgweg 9a 53115 Bonn (Deutschland)

Andreas Strub, Karin Röttgers, Wolfgang Voos The peptide-binding domain determines the interaction of mitochondrial Hsp70s with the essential partner protein Tim44 The import of cytosolic preproteins into the mitochondrial matrix is driven by the ATP-dependent activity of an import motor complex located at the inner face of the inner membrane. The core component of this complex is the Hsp70 chaperone Ssc1. It is anchored to the inner membrane by the interaction with the membrane translocase component Tim44. As a third component, the nucleotide exchange factor Mge1 enhances the ATP activity of Ssc1. In the mitochondrial matrix two other members of the Hsp70 family have been identified. However, despite an extremely high amino acid sequence conservation, Ssq1 and Ssc3 (Ecm10) are not able to replace the essential function of Ssc1. By assaying protein interactions in mitochondria we found that neither Ssq1 nor Ssc3 were able to interact with the essential partner protein Tim44 although all three Hsp70s compete for the nucleotide exchange factor Mge1. To address the functional correlation between Ssc1, Ssq1 and Ssc3, we performed a comprehensive analysis of the behavior of Hsp70 domain exchange constructs in vitro and in vivo. In the full length protein context, chimeric proteins that contained the peptide-binding domain of Ssc1 were found to interact with Tim44. However, the single peptide-binding domains were not able to interact with Tim44 while the ATPase domains of all three mitochondrial Hsp70s did bind specifically to Tim44. A fully functional domain interaction was found to be essential for the translocation function of Ssc1. We conclude that the presence of the peptide binding domains determines the binding behavior of the respective ATPase domain with Tim44, suggesting a new regulatory principle of Hsp70 partner protein interaction. Literature Voisine, C. et al. (1999). Cell 97, p. 565 Krimmer T. et al. (2000). Mol. Cell Biol. 20, p. 5879 Lim, J. H. et al. (2001). EMBO J. 20, p. 941 Schmidt S. et al. (2001). J. Mol. Biol., 313, p. 13 Strub, A. et al. (2002). EMBO J., in press contact: PD Dr. Wolfgang Voos Universität Freiburg Institut für Biochemie [email protected] Hermann-Herder-Str. 7 79104 Freiburg (Germany)

Doris Kloor1, Angelika Lüdtke1, Stanka Stoeva2, Hartmut Osswald1 The ratio of the cofactor NAD +/NADH of S-adenosylhomocysteine hydrolase controls the adenosine binding sites. S-adenosylhomocysteine (SAH) hydrolase catalyzes the reversible hydrolysis of SAH to adenosine (Ado) and homocysteine and regulates S-adenosylmethionine dependent transmethylation reactions. One mol of SAH hydrolase contains 4 mol tightly bound NAD+. Ado leads to a gradually reduction of the tightly bound NAD+/ to NADH in the SAH hydrolase. In the present investigation we examined the Ado binding characteristics of modified SAH hydrolase with defined NAD+/NADH ratios. Methods: SAH hydrolase was purified to homogeneity from bovine kidney. To prepare the enzyme with defined NAD+/NADH ratios, bound NAD was removed by dialysis and the apo-enzyme was reconstituted with NAD+/NADH ratios between 1/0 and 0/1. For saturation binding experiments the enzyme (10 mg/ml) was incubated with 3H-Ado (0.01-60 µmol/l) over night at 4°C in Tris/Hepes pH 7.4. To identify the Ado binding sites, SAH hydrolase was incubated with [2-3H]-8-azido-Ado. After irradiation the reaction mixture was digested by Asp-N or trypsin and the peptides, purified by HPLC, were sequenced. Results: SAH hydrolase in its native form and at a ratio of NAD+/NADH 0.5/0.5, exhibits two binding sites for Ado with a KD1 of 9.2±0.6 nmol/l and a KD2 1.4±0.1 µmol/l, respectively, which correspond to peptides of native SAH hydrolase identified as Asp307-Val325 and Tyr379-Thr410. Binding of Ado to SAH hydrolase with NAD+/ NADH ratios of 1/0 and 0/1 revealed only one binding site with low affinity of 4.9±0.3 µmol/l and high affinity of 48.2±2.7 nmol/, respectively. Only one photolabelled peptide was identified as Asp391-Ala396 from the NAD+ form and as Asn312-Lys318 from the NADH form of the enzyme. Conclusion: Our data show that SAH hydrolase has two different binding sites for Ado. This finding may help to elucidate the enzymatic mechanism. Depts. of 1Pharmacology, Faculty of Medicine and 2Physical Biochemistry, Eberhard-Karls Universität, 72074 Tübingen, Germany contact: Dr. Doris Kloor Universität Tübingen Institut für Pharmakologie [email protected] Wilhelmstrasse 56 72074 Tübingen (Deutschland)

Jörg Tatzelt, Konstanze F. Winklhofer The role of co- and posttranslational modifications in the propagation of scrapie-prions A hallmark of mammalian prion diseases is the conversion of the cellular prion protein PrPC into an abnormally folded isoform, designated PrPSc, which represents the major component of infectious prions. During transit through the secretory pathway the C-terminal domain of PrP is posttranslationally modified by the attachment of two N-linked complex carbohydrate moieties and a glycosylphosphatidylinositol (GPI) anchor at the C terminus. Glycosylation of the prion protein is implicated in neuronal targeting of PrPSc and the phenomenon of prion strain diversity. Here we show that inhibition of terminal glycosylation promotes propagation of PrPSc and creates a new glycotype pattern. Interestingly, geldanamycin, a hsp90/grp94-binding drug, interfered with glycan processing in a similar manner to α-mannosidase inhibitors. Moreover, in geldanamycin-treated cells grp94 was found in complex with PrP containing high mannose N-linked glycans. Our study reveals that glycan processing in the secretory pathway modulates PrPSc propagation and the signature of prion strains. contact: Dr. Konstanze F. Winklhofer Max-Planck-Institut fuer Biochemie [email protected] Am Klopferspitz 18a 82152 Martinsried (Germany)

Seigo Izumo The role of Csx/Nkx 2.5 homeobox gene in cardiac development and congenital heart disease. Accumulating evidence indicates that the molecular pathway of cardiogenesis is highly conserved throughout evolution. The Drosophila tinman gene is a homeobox transcription factor essential for early heart development in Drosophila. Csx (or Nkx 2.5) is a murine homologue of tinman and is initially expressed in the precardiac mesoderm and the pharyngeal endoderm . The null mutation of Csx/Nkx 2.5 in the mouse causes the arrest of heart development soon after completion of the looping of the heart tube. Interestingly, heterozygous mutations of the human Csx/Nkx 2.5 were shown to cause a variety of congenital heart disease, including atrial septal defect (ASD), AV conduction block, tetralogy of Fallot, Ebstein s anomaly, and left ventricular noncompaction. Heterozygous mutations of Csx/Nky 2.5 in the mouse cause ASD, AV conduction block, and vulnerability to arrhythmia. Targeted deletions of specific regions of Csx/Nkx 2.5 have indicated that the evolutionarily conserved domains serve essential functions at different stages of cardiac development. Tix/Nkx 2.6 is another murine homologue of tinman. It is expressed in the pharyngeal endoderm and in the cardiac outflow tract. Targeted mutation of Tix/Nkx 2.6 has revealed no obvious phenotype. However, double knockout of Tix/Nkx 2.6 and Csx/Nkx 2.5 caused a severe pharyngeal defect and cardiac abnormalities. Together with the data obtained in the round worm C elegance, the evolutionary origin of the heart is the upper gastrointestinal muscle, rather than the body wall muscle. Thus, study of the transcription factor Csx/Nkx 2.5 family of genes has provided new insight into the evolutionary history of the heart as well as the molecular basis of congenital heart disease in humans. contact: Professor Seigo Izumo Harvard Medical School Beth Israel Deaconess Medical Center [email protected] 330 Brookline Avenue MA 02245 (USA)

Garret A. Fitzgerald The role of cyclooxygenases in the cardiovascular system Habermann Lecture 2002 contact: Dr. Garret A. Fitzgerald University of Pennsylvania School of Medicine 3620 Hamilton Walk Philadelphia, PA (USA)

Simone Diestel, Claudia Müller, Marie-Therese Fergen, Brigitte Schmitz The role of NCAM phosphorylation on NCAM mediated signal transduction pathways The neural cell adhesion molecule NCAM is a posttranslational modified protein with multiple phosphorylation sites located in the cytoplasmic tail. To study the role of intracellular phosphorylation of human NCAM, one potentially phosphorylated amino acid was exchanged against an amino acid which can not be phosphorylated. The neuronal rat B 35 cell line which shows low endogenous NCAM expression was stably transfected with wild type and mutant human NCAM 140 and NCAM 180 cDNA, respectively. Positive clones with nearly identical expression levels were chosen for further analysis. We compared the phosphorylation levels of wild type and mutant proteins and measured the effects of the mutation on different signal transduction pathways and neurite outgrowth. contact: Dr. Simone Diestel Universität Bonn Institut für Physiologie, Biochemie und Hygiene der Tiere [email protected] Katzenburgweg 9a 53115 Bonn (Deutschland)

Martina Schultheis, Brigitte Schmitz The role of serine-phosphorylation of the cell adhesion molecule L1 in basal neurite outgrowth The cytoplasmic domain of the cell adhesion molecule L1 is highly conserved between species and known to be phosphorylated by a few specific kinases. To investigate the significance of L1 phosphorylation for basal (i.e. unstimulated) neurite outgrowth mutants of human L1were generated, in which several known phosphorylated serine residues were converted into non phosphorylatable amino acids. Neuronal B35 rat cells, which have a very low endogenous L1 expression, were stably transfected with the L1 constructs and positive clones were identified by indirect immunofluorescence and Western blot analysis. Neurite outgrowth of B35 cells expressing similar levels of wild type or mutated L1 was determined under different condititons. The results show that L1 phosphorylation can directly influence basal neurite outgrowth. Further investigations into underlying signaling mechanisms are being carried out to understand the functional role of L1 phosphorylations in basal neurite outgrowth. contact: Martina Schultheis Universität Bonn Institut für Physiologie, Biochemie und Hygiene der Tiere [email protected] Katzenburgweg 9a 53115 Bonn (Deutschland)

Stefanie Barbirz The tailspike from E.Coli H bacteriophage HK620: Another Parallel beta-Helix Protein? Recently, sequencing of the E.Coli H phage HK620 resulted in cloning of the gene for its tailspike protein, HK620 TSP. The first about 100 amino acids of the 710 residue protein exhibit high sequence homology in its N-terminal capsid binding domain with the tailspikes of two other phages of the Podoviridae family, Salmonella phage P22 and Shigella phage Sf6. However, sequence homology between the three tailspikes is completly lacking in the major C-terminal part which forms a large parallel-b-helix domain in P22 TSP. In spite of the completely differing sequences we propose the same parallel b-helix fold for HK620 TSP. To support this, we present here the spectroscopical and biochemical characterization of HK620 TSP. The protein was recombinantly expressed in E.coli and purified. HK620 TSP is a SDS-resistant thermostable trimer with an molecular weight of 230 kDa as shown by size-exclusion chromatography. The secondary structure of the protein is similar to that of P22 TSP as shown by circular dichroism and FT-IR spectroscopy. The N-terminal capsid binding domain can be deleted without affecting the ability of the protein to form a trimer. This truncated protein is resistant to proteases and shows thermostability comparable to P22 TSP. It was predominantly used for crystallization experiments. Conclusive evidence for the similarity of the two tailspike structures can only be provided by crystallographic data once available. However, biochemical and spectroscopical characterization strongly suggest that HK620 tailspike protein joins the family of trimeric parallel beta-helix-proteins as a third member together with P22 and Sf6 tailspikes. contact: Stefanie Barbirz Universität Potsdam Biologie und Biochemie [email protected] Karl-Liebknechtstr. 24-25, Haus 25 14476 Golm (Deutschland)

Peter Rehling, Peter Kovermann, Richard Wagner, Nikolaus Pfanner, Kaye, N. Truscott, Katrin Brandner The TIM22 translocase mediates insertion of carrier precursors into the mitochondrial inner membrane via a two-step process The vast majority of mitochondrial proteins are nuclear-encoded and synthesized in the cytosol. Depending on their final destination their import may require translocation machinery present in both the outer and inner membranes. Specific targeting signals, contained within preproteins, direct them to the mitochondrial surface where they are imported into or through the lipid bilayer by the translocase of outer membrane (TOM). Two distinct translocases of the inner mitochondrial membrane (TIM), facilitate import of preproteins into or across this internal lipid bilayer. Preproteins with positively charged presequences at their N-terminus are inserted via the TIM23 complex, whereas preproteins with internal targeting information such as carrier proteins are imported via the TIM22 complex. In both cases a membrane potential across the inner membrane is essential for their import. Carrier preproteins are imported into mitochondria in five defined stages: stage I – binding to cytosolic chaperones; stage II – recognition by receptors on the outer membrane surface; stage III –transport through the outer membrane; stage IV – insertion into the inner membrane; stage V – dimerisation. At stage IV carrier preproteins are directed to the TIM22 complex (formed by Tim54, Tim22, Tim18 and small Tims) where insertion into the inner membrane is mediated by the pore forming protein Tim22. We have examined the requirements for stage IV import using the functionally intact purified TIM22 complex that associate in a stable manner with carrier translocation intermediates. Electrophysiological measurements with the isolated TIM22 complex and in vitro import assays with carrier preproteins together indicate that carrier membrane insertion is a two-step process intimately linked to the energy source, the membrane potential. contact: Dipl.-Ing. (FH) Katrin Brandner Universität Freiburg, Pfanner Institiut f. Biochemie und Molekularbiologie [email protected] Hermann-Herder 7 79104 Freiburg (Deutschland) additional information Brandner, K.1, Rehling, P.1, Kovermann, P.2, Wagner, R.2, Pfanner, N.1 & Truscott, K. N.1 1Institut für Biochemie und Molekularbiologie, Universität Freiburg, Hermann-Herder-Strabe 7, D-79104 Freiburg, Germany, 2Biophysik, Universität Osnabrück, FB Biologie/Chemie, D-49034 Osnabrück, Germany.

Steffen Liedtke The Use of Multidimensional Capillary LC in Proteomics The Use of Multidimensional Capillary LC in Proteomics Steffen Liedtke (LC Packings – A Dionex Company, Idstein, Germany) After completing a large number of genome sequences during the last decade attention is now focused on the structure and function of proteins in biological systems. For more than two decades, commercial 2D electrophoretic techniques have provided the highest resolving power available. Large-scale protein profiling is generally performed by two-dimensional gel electrophoresis (2D GE) followed by MS or LC-MS. The limitations of the separation method, i.e. 2D GE, however, are by now well documented (e.g., the identification of low-abundance and/or water insoluble [membrane] proteins is problematic). Chromatographic techniques, in particular Capillary- and Nano LC, coupled to electrospray mass spectrometry (ESI-MS) are increasingly applied for the analysis of complex peptide samples. However the peak capacity of a typical Nano LC run is between 50 and 75 peaks. Although modern MS instruments can fractionate multiple components simultaneously, still the cycle time is limiting the amount of information when too many peaks are co-eluting. Therefore, the analysis of very complex samples requires more resolving power of the chromatographic system. In recent years, the advantages of two-dimensional methods based on liquid chromatography have become evident. In this application a FAMOS micro autosampler, a UltiMate Nano LC system and a Switchos microcolumn switching unit are used for 2D LC analysis of a complex peptide mixtures. The method is based on fractionation of peptides on a strong cation exchange stationary phase, desalting the petides on a reversed phase precolumn and separation on a reversed phase Nano LC column prior to the LC-MS analysis. contact: Dr. Steffen Liedtke LC Packings - A Dionex Company [email protected] Am Wörtzgarten 10 65510 Idstein (Deutschland)

Simone Fulda, Klaus-Michael Debatin Therapeutic modulation of death receptor- or drug-mediated apoptosis Apoptosis, the cell’s intrinsic death program, plays a crucial role in the regulation of tissue homeostasis, and an imbalance between cell death and proliferation may result in tumor formation. Also, killing of tumor cells by diverse cytotoxic approaches such as anticancer drugs, irradiation, death receptor ligands, suicide genes or immunotherapy, is predominantly mediated through induction of apoptosis in tumor cells. Activation of the cascade of proteolytic enzymes known as caspases is a critical component of the execution phase of cell death in most forms of apoptosis. Two main caspase cascades, one triggered by death receptor stimulation and the other one initiated at the mitochondria, have been identified in response to various inducers of cellular stress such as DNA damage. Activation of caspases and apoptosis is tightly regulated at several levels, e.g. by Bcl-2 family proteins, by inhibitor of apoptosis proteins (IAPs) and upstream inhibitors such as FLIP. Failure to activate apoptotic pathways in response to drug treatment may lead to resistance of tumors cells to anticancer therapies. Therefore, factors affecting caspase activation and apoptosis might be important determinants of drug sensitivity. In addition to caspase-dependent apoptosis, caspase-independent forms of cell death may also play a role for treatment response. Insights into the mechanisms regulating apoptosis as well as other forms of cell death pathways provide a molecular basis for novel strategies targeting resistance of tumor cells. contact: PD Dr. Simone Fulda Universität Ulm Kinderklinik [email protected] Prittwitzstr. 43 89075 Ulm (Germany)

Tanja Richter, Kathrin Klein, Thomas E. Mürdter, Michel Eichelbaum, Matthias Schwab, Ulrich M. Zanger ThioTEPA and Clopidogrel are specific mechanism-based inhibitors of human CYP2B6 Human CYP2B6 catalyzes the metabolism of several drugs including cyclophosphamide and the antidepressant, bupropion. Investigation of this enzyme has long been hampered by the lack of suitable probes. The cytostatic ThioTEPA was recently shown to inhibit cyclophosphamide hydroxylation in vivo whereas the platelet aggregation inhibitor, clopidogrel, was suggested to be a substrate of CYP2B6. By measuring bupropion-hydroxylation in liver microsomes and in recombinant P450s, we found that these two substances strongly inhibit CYP2B6 in a time- and concentration-dependent manner by up to 90% and with IC50 values of about 5 and 0.5 µM, respectively. As shown by dialysis- and microfilter experiments, inhibition was irreversible and dependent on the presence of NADPH, strongly suggesting a mechanism-based mode of action for both thioTEPA and clopidogrel. High specificity of both inhibitors for CYP2B6 was demonstrated by analyzing enzyme activities selective for other human liver CYPs including ethoxycoumarin-O-dealkylation (CYP1A2 and CYP2E1), propafenon 5-hydroxylation (CYP2D6), coumarin 7-hydroxylation (CYP2A6), verapamil N-demethylation (CYP3A4/5), verapamil O-demethylation (CYP2C8), S-mephenytoin N-demethylation (CYP2C9) and S-mephenytoin 4'-hydroxylation (CYP2C19). These results suggest the possibility of drug interactions between these two drugs and other drug-substrates of CYP2B6. Clopidogrel in particular may prove to be a useful in-vitro probe for this enzyme. Supported by the DFG and by the Robert Bosch Foundation, Stuttgart. contact: Tanja Richter Dr. Margarete Fischer-Bosch Institut für Klinische Pharmakologie [email protected] Auerbachstr. 112 70376 Stuttgart (Deutschland)

Thomas Dandekar Tools of bioinformatics: Comparative genome analysis and pathway reconstruction Pathway reconstruction builds on genome analysis and biochemical data with the aim of reconstructing higher level interactions between identified enzymes in a specific genome (1). Comparative genome comparisons exploit approaches such as differential genome analysis. Building on this and further results different enzyme pathways are compared and reconstructed. Metabolite flow in a pathway is analyzed by further tools such as elementary mode analysis. An overview of bioinformatic tools and algorithms for these different tasks is presented. They reveal key enzymes and pharmacological targets in the enzyme network. Target selection and drug development can both be sped up by this. Literature (1) Dandekar T, Sauerborn R. (2002) Comparative genome analysis and pathway reconstruction. Pharmacogenomics. 3(2):245-256 contact: Prof. Dr. Thomas Dandekar Wuerzburg Bioinformatik [email protected] Biozentrum, Am Hubland 97074 Wuerzburg (Germany)

Franz Oesch Toxicology from experimental systems to man: drug metabolising enzymes as key determinants Extent and quality of the xenobiotic metabolism differ drastically depending on many factors such as the species differences in metabolism, that pose the severe problem of limited transferability of results obtained in a toxicological experiment with a model animal to man. Thus, it is evident that extensive knowledge on the differences in metabolism pathways between human and the model species is a prerequisite to extrapolate the obtained results in a meaningful way. Moreover, there are the inherent characteristics of a given individual, such as sex, age, and genetic predisposition, that represent important determinants of xenobiotic metabolism. Finally, environmental factors strongly modulate the expression and activity of xenobiotic metabolising enzymes. Chemical compounds themselves are often capable to induce, repress, inhibit or activate these enzymes. Nutritional behaviour and dietary ingredients substantially modulate the metabolic competence. Disease states, such as infections, chronic liver disease or simply fever, can selectively alter the capacity of defined xenobiotic metabolising enzymes significantly. Metabolic activation is a prerequisite for the genotoxicity of most chemical mutagens and carcinogens. Reactive metabolites are under the control of activating, inactivating and precursor sequestering enzymes. Such enzymes themselves are under the long-term control of induction and repression as well as under the short-term control of posttranslational modification. The latter is especially effective since it works fast and in many cases affects the enzymatic activity as well as the rate of degradation of the enzyme itself. Moreover, the compartmentalization of these systems and of the target molecules play a crucial role in the enzymatic control of toxicity. All these enzymes differ widely between available toxicological test systems, animal species and man. However, correct predictions of genotoxic risks are especially important because of the long latency time of genotoxic effects. Therefore, careful consideration of the basic mechanisms responsible for the control of the ultimately active species derived from genotoxic tumor initiators is very important for toxicological risk evaluations. contact: Prof. Dr. Franz Oesch Universität Mainz Institut für Toxikologie [email protected] Obere Zahlbacher Str. 67 55131 Mainz (BRD)

S. Breidert, R. Jacob, A. Ngezahajo, H.-A. Kolb, H. Naim Trafficking pathways of connexin49-GFP in living mammalian cells The present study examined the trafficking pathways of connexin49 (Cx49) labelled with green fluorescent protein (GFP) in polarized and nonpolarized eukaryotic cell lines. Connexin proteins are thought to follow several transport pathways including subcellular compartments as well as cytosolic transport routes towards the plasma membrane. There, they are integrated to form gap junction plaques between adjacent cells. Cx49 was cloned from ovine lens by RT-PCR. The Cx49 cDNA sequence was fused in frame to GFP (povCx49-GFP) and transfected into several eukaryotic cell lines. Synthesis, assembly, degradation and removal of povCx49-GFP was determined by pulse-chase experiments in nonpolar COS cells. The trafficking pathways were monitored by confocal microscopy. Fusion proteins were localized in subcellular compartments including ER, ERGIC, Golgi, trans Golgi network (TGN)as well as in transport vesicles travelling towards the plasma membrane. Gap junction plaque assembly resembling the classical punctuate distribution pattern could be demonstrated for nonpolar COS-7 and polar MDCK cells. A basolateral distribution of povCx49-GFP became apparent in polar MDCK cells indicating a specific sorting behaviour for polar organized cells. The results gained by confocal microscopy and pulse-chase experiments provide a conclusive evidence for a transport competent conformation of povCx49-GFP in all examined cell lines. contact: Prof. Dr. Hassan Naim Tierärztliche Hochschule Hannover Institut für Physiologische Chemie [email protected] Bünteweg 17 30559 Hannover (Deutschland) additional information PD Dr. R. Jacob Institut für Physiologische Chemie Tierärztliche Hochschule Hannover Bünteweg 17 e-mail:[email protected] Prof. Dr. H.-A. Kolb Dr. A. Ngezahajo Dipl.-Bio. S. Breidert Institut für Biophysik Universität Hannover e-mail: [email protected]

Markus von Nickisch-Rosenegk, Jenny Steffen, Frank F. Bier Transcription of Reporter Genes with Immobilized Templates Our first attempt regarding the reconstruction of cellular mechanisms on biochips (e.g. proteine synthesis) was the construction and the immobilization of transcriptionable DNA. Therefore we chose the EGFP gene which could report itself in further translation experiments. In a PCR reaction we amplified the flourescent protein gene using a forward primer which contains a T7 promotor region followed by a Shine-Delgardo-motiv upstream to the startcodon of the EGFP gene primer sequence and a fluorescein-modification (FAM) at the 5‘-end. The reverse primer was modified with biotin at its 5‘-end and has a poly-T spacer linked to the stopcodon of the gene. The amplicons were spotted on a neutravidin-covered glaschip and immobilized via the biotin molecule of the reverse primer. After a wash step the spots could be verified by UV excitation an the resulting fluorescence of the FAM molecule. These fixed DNA molecules were incubated with a reaction mixture containing T7-RNA-Polymerase for in vitro on-chip-transcription. The synthesized RNA was verified by DNA digestion using DNAse I followed by RT-PCR or by an incorporation of FITC-dUTP during the RNA synthesis. A third method was an incorporation of Cy5 dye-labeled dNTPs during a first strand cDNA synthesis. contact: Dr. Markus von Nickisch-Rosenegk Fraunhofer Gesellschaft IBMT [email protected] Arthur-Scheunert-Allee 114-116 14558 Bergholz-Rehbrücke (D)

Michael Metzlaff Transgene- and virus-mediated gene silencing - What plants can teach us! First observations that the addition of extra copies of genes can cause silencing of gene expression were published for transgenic petunia plants in 1990 (1,2). The transformation of petunias with transgenes coding for chalcone synthase, which is the key enzyme in flower pigmentation, did surprisingly result in plants with partial or total loss of flower color instead of plants with an expected enhancement in flower pigmentation. Because both types of genes, the endogenous chalcone synthase genes and the added transgenes, became silenced, the phenomenon was called co-suppression. After these initial observations, numerous cases in plants and shortly later in fungi, worms, insects and mammals revealed that the phenomenon seems to occur in all higher organisms and that it can act at the transcriptional and post-transcriptional level of gene expression. It was also demonstrated that the infection of plants with viruses can induce the same gene silencing pathways (3), whereof it is now assumed that the major biological role of gene silencing in eukaryotic cells consists in protection against invading foreign nucleic acids. The intense effort for tracing the molecular mechanisms acting in gene silencing led within a few years to the discovery of a number of new RNA species resulting from and interacting with gene silencing. Based on own research data (4,5) and those from literature I will draw a picture of an interactive network including all so far identified RNA species, i.e. aberrant sense and antisense RNAs, double stranded RNAs, small interfering and small regulatory RNAs. Double stranded RNAs turned out to be the central molecules for the induction of gene silencing (6). Several mutants with deficiencies in gene silencing gave first hints for which proteins play a role in gene silencing. I will position also these proteins within the interactive network. Today gene silencing is not only of academic interest but has already developed into a powerful genomics tool used for gene function discovery and validation. In animal and human cell systems, transient and recently also transgene-based RNA interference (RNAi) has been widely applied for gene-specific inhibition of gene expression (6,7). In plants, transgenes with inverted repeat structures (8) and chimeric viruses (9 and own results) have been exploited for down-regulating the expression of endogenous genes involved in various biochemical pathways. The 12-years "history" of gene silencing is another example that basic research in a specific field of biology, in this case in the plant field, can have an enormous influence on the direction and speed of developments in other research domains. Thus, silenced plants have not only given us exciting insights into novel mechanisms of gene expression control but have also taught us once more, how important it is in modern sciences to develop an interactive research culture without barriers between research domains. Literature (1) Napoli, C., Lemieux, C. & Jorgensen, R. Introduction of a chimeric chalcone synthase gene into petunia results in reversible co-suppression of homologous genes in trans. Plant Cell 2, 279-289 (1990). (2) Van der Krol, A.R., Mur, L.A., Beld, M., Mol, J.N.M. & Stuitje, A.R. Flavonoid genes in petunia: addition of a limited number of gene copies may lead to cosuppression of gene expression. Plant Cell 2, 291-299 (1990). (3) Lindbo, J.A., Silva-Rosales, L., Proebsting, W.M. & Dougherty, W.G. Induction of a highly specific antiviral state in transgenic plants: Implecations for regulation of gene expression and virus resistance. Plant Cell 5, 1749-1759 (1993). (4) Metzlaff, M., O'Dell, M., Cluster, P.D. & Flavell, R.B. RNA-mediated RNA degradation and chalcone synthase A silencing in petunia. Cell 88, 845-854 (1997). (5) Metzlaff, M., O'Dell, M., Hellens, R. & Flavell, R.B. Developmentally and transgene regulated nuclear processing of primary transcripts of chalcone synthase A in petunia. Plant J. 23, 63-72 (2000). (6) Fire, A., Xu, S., Montgomery, M.K., Kostas, S.A., Driver, S.E. & Mello, C.C. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391, 806-811 (1998). (7) Paddison, P.J., Caudy, A.A., Bernstein, E., Hannon, G.J. & Conklin, D.S. Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells. Genes & Develop. 16, 948-958 (2002). (8) Waterhouse, P.M., Graham, M.W. & Wang, M.B. Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA. Proc. Natl. Acad. Sci. USA 95, 13959-13964 (1998). (9) Baulcombe, D.C. Fast forward genetics based on virus-induced gene silencing. Curr; Opin. Plant Biol. 2, 109-113 (1999). contact: PD Dr. habil. Michael Metzlaff Aventis CropScience N.V. [email protected] Jozef Plateaustraat 22 9000 Gent (Belgium)

Dirk H. Ostareck Translational activation from DICE-mediated mRNA silencing by the Src tyrosine kinase Dirk H. Ostareck, Antje Ostareck-Lederer, Christophe Cans, Gitte Neubauer, Karol Bomsztyk* , Giulio Superti-Furga, Matthias W. Hentze EMBL, Meyerhofstr. 1, 69117 Heidelberg, Germany, *Dept. of Medicine Univ. of Washington, 98195 Seattle,WA, USA Reticulocyte-15-lipoxygenase (15-LOX) is a key enzyme in erythroid cell maturation. 15-LOX catalyses the dioxygenation of intact phospholipids in mitochondrial membranes and participates in their breakdown in mature reticulocytes. The 15-LOX mRNA is transcribed in bone marrow erythroblasts and its translation is silenced until enucleated reticulocytes in the peripheral blood undergo the final stages of maturation. Silencing is achieved via a motif in the 3’UTR of 15-LOX mRNA, the Differentiation Control Element (DICE). We purified two proteins that bind to the DICE, hnRNPs K and E1, which silence 15-LOX mRNA translation specifically in vitro and in vivo (1, 2). Here we address the question of how the silenced mRNA can be translationally activated. We show that hnRNP K and c-Src kinase specifically interact with each other, leading to c-Src activation and tyrosine phosphorylation of hnRNP K in vivo and in vitro. c-Src mediated phosphorylation reversibly inhibits the binding of hnRNP K to the DICE in vitro, specifically de-represses the translation of DICE-bearing mRNAs in vivo. An hnRNP K mutant that cannot be phosphorylated by c-Src functions as a Src-resistant translational repressor in vivo. The timing of c-Src expression during erythroid differentiation implicates c-Src in the activation of the LOX mRNA in maturing red blood cells(3). Literature 1)Ostareck D.H., Ostareck-Lederer A., et al., Cell 1997 (89) 597-606. 2)Ostareck D.H., Ostareck-Lederer A., et al., Cell 2001 (102) 281-290. 3)Ostareck-Lederer A., Ostareck, D.H., et al., Mol. Cell. Biol. 2002 (22) 4499-4511. contact: Dr. rer. nat. Dirk H. Ostareck EMBL [email protected] Meyerhofstr. 1 69117 Heidelberg (Germany) additional information http://www.embl-heidelberg.de/ExternalInfo/hentze/Dirkpage.html

Sabine Molik, Ralf Bernd Klösgen Transport and assembly of the Rieske Fe/S protein: What happens in the stromal space? The Rieske Fe/S protein is a nuclear-encoded subunit of the cytochrom b6/f complex in chloroplasts. It is synthesised in the cytosol as a precursor molecule with an N-terminal transit peptide, post-translationally transported into the organelle and subsequently targeted via the delta pH/TAT pathway to the thylakoids. In contrast to all other thylakoid proteins analysed so far, transport of the Rieske protein is retarded in the stromal space leading to the accumulation of the protein in several complexes of high molecular weight. Using various chimeric and mutant Rieske proteins, complexes interacting specifically with either the hydrophilic lumenal domain or the thylakoid targeting signal which is provided by the membrane anchor can be distinguished. Among the latter is the cpn60 system, the well known folding machinery in the chloroplast. Other stromal complexes might be responsible for Fe/S cofactor insertion, because delta pH/TAT pathway is capable of transporting folded proteins. In the case of the Rieske protein, correct folding and/or assembly of the iron-sufur cluster seems even to be a prerequisite for translocation across the thylakoid membrane. contact: Sabine Molik Martin-Luther-Universität Halle-Wittenberg Institut für Pflanzenphysiologie [email protected] Weinbergweg 10 06120 Halle (Saale) (Germany)

Joerg W. Bartsch, Dirk Wildeboer, Uwe Schlomann Tumor necrosis factor alpha signalling in reactive glia cells As a response to neuronal injury or cell death, glia cells, astrocytes and microglia, are activated, thereby changing the expression of many genes. In neurodegenerative diseases, reactive glia cells are able to remodel the extracellular matrix, forming a glial scar and produce a number of cytokines, among them Tumor Necrosis Factor (TNF), TNF-&alpha. Some target genes of TNF-&alpha in reactive glia cells were described, but their impact for the progression of CNS pathology remained unclear. One target gene is ADAM8, a metalloprotease-disintegrin, that is upregulated in reactive glia cells by TNF-&alpha in the Wobbler mouse model for neurodegeneration (1). The signalling pathway of ADAM8 transcriptional activation by TNF-&alpha was analysed in human cell lines 1321N1 (astrocytoma) and in hMC3 (microglia). In both cell lines, TNF-&alpha leads to transcriptional activation of Interferon Regulatory Factor-1 (IRF-1) by TNFRI signalling. IRF-1 activates ADAM8 transcription. The signalling pathway leading to IRF-1 transcription in both cell lines was different: whereas in astrocytoma cells, ADAM8 transcription was suppressed by AG490, an inhibitor of the JAK/STAT pathway, signalling in microglia cells was strongly dependent on NF-&kappaB, as demonstrated by complete inhibition of ADAM8 transcription with pyrrolidine dithiocarbamate, a selective inhibitor of NF-&kappaB. NF-&kappaB is well-documented as transcription factor related to CNS pathology. In contrast, our results demonstrate that IRF-1 seems to play an immunomodulatory role in reactive astrocytes and among the target genes of IRF-1, genes with neurotrophic properties were found, assuming that TNFRI-mediated TNF-&alpha activation of the IRF-1 pathway might serve neuroprotective roles. Supported by DFG, SFB 549. Literature (1) Schlomann, U., Rathke-Hartlieb, S., Yamamoto, S., Jockusch, H., Bartsch, J.W.(2000) J. Neurosci. 20, 7964-7971.

contact: Dr. Joerg W. Bartsch Universität Bielefeld Entwicklungsbiologie und Molekulare Pathologie [email protected] Universitätstr. 25 33615 Bielefeld (D)

Albert Sickmann, Helmut E. Meyer, Yvonne Wagner Two-dimensional chromatography in protein analysis The analysis of complex protein mixtures like ribosomes, centrosomes, peroxisomes and further multi protein complexes is still a difficult problem. In general such analyses are done by a high-resolution technique, e.g. 2D gel electrophoresis (2D-PAGE), followed by a protein identification via mass spectrometry. The 2D-PAGE is a time-consuming method for the separation and is not suitable for all kinds of proteins, e.g. hydrophobic proteins, basic proteins, very large or very small proteins. To overcome this problem we validated methods for the analysis of complex protein mixtures that manage with very small amounts of protein. The separation is done either with 2D-nano-HPLC or 1D-nano-HPLC. Both methods were validated by separation of yeast-ribosomes. The amount of the digested ribosomes was less than 1 µg. For 2D-HPLC an orthogonal system was used consisting of a capillary ion exchange chromatography for the first dimension and a nano-reversed phase chromatography for the second one. Between both dimensions the sample is cleaned up by a preconcentration step. Furthermore, the reversed phase column was coupled with an ion trap mass spectrometer (ESI-MS) recording the mass spectra automatically. A comparison between the two-dimensional chromatography and a one-dimensional separation showed a larger sum of identified proteins and furthermore a better sequence coverage within the two-dimensional system using the same protein amount. In addition the possibility of combining different one-dimensional separation methods adapt this technique to almost any biological sample. contact: Yvonne Wagner Ruhr-Universität Bochum Medical Proteom-Center [email protected] Universitätsstr. 150 44780 Bochum (Germany)

Andrè Reis Understanding the genetic basis of common human diseases and its potential for new concepts in drug therapy contact: Prof. Andrè Reis Universität Erlangen [email protected] Erlangen (D)

Gunter Fischer Unfolding the Chemistry of Protein Folding Protein folding is the process by which the polypeptide backbone can be thought of as undergoing simple motions like torsional movements and chain diffusion. There appears at first glance little evidence of factors other than physical determinants that are able to dominate protein folding and backbone reorganization. Whether chemical events like catalysis are possibly involved relate to the magnitude of bond order changes in approaching the transition state of folding. The traditional picture of the amide resonance illustrates the bond order changes implicated in nitrogen rehybridization. Consequently, geometric isomers exist, where a high energy barrier separates two ground states per peptide bond in all folding states of a polypeptide (1). If the interconversion of the isomers becomes rate-limiting in folding and biorecognition, isomer-specific reactions result as could be found for proteolysis, protein phosphorylation/dephosphorylation, ligand-receptor interactions and chain rearrangements in protein folding. Typically, the relaxation rate for secondary amide peptide bond isomerization was found on the order of one second, whereas peptidyl prolyl imide bonds interconvert on a time scale of minutes. Had isomerization rates been too slow, nature would have evolved enzymes for specific acceleration. The characterization of the hsp70 chaperone DnaK as the first member of the secondary amide peptide bond cis/trans isomerases (APIases)(2), and biochemical functions of members of the currently existing families of the peptidyl prolyl cis/trans isomerases (cyclophilins, FK-506 binding proteins and parvulins, PPIases) will lead toward a generalization of the folding control mediated by chemical properties of the protein backbone. Literature (1) U. Reimer and G. Fischer Biophys. Chem. 96, 203-212 (2002). (2) C. Schiene-Fischer, J. Habazettl, F. X. Schmid and G. Fischer Nature Struct. Biol. 9, 419-424 (2002) contact: Prof. Dr. Gunter Fischer Max Planck Forschungsstelle für Enzymologie der Proteinfaltung [email protected] Weinbergweg 22 06120 Halle (D)

Annette Deul, Susanne Stevens, Uta Sander, Christian Kirchhoff, Joern Jungmann, Thomas Hesterkamp Use of cytochrome P450 isoenzyme assays in the hit-to-lead process of drug discovery Drug-drug interactions at the level of cytochrome P450 (CYP) mediated drug metabolism are a frequent cause of failure of clinical candidates. Of outstanding medical relevance are the human CYP isoenzymes 1A2, 2C9, 2C19, 2D6, and 3A4. In order to routinely test inhibition of these isoenzymes during early stages of drug discovery, i.e. the hit-to-lead process, available fluorogenic assay principles were adapted to a medium throughput 384 well assay format. The novel assay format was validated using (i) known standard inhibitors for these isoenzymes and (ii) a library of 560 synthetic compounds made up of 20 different structural scaffolds with unknown interference with CYP activity. The good assay performance allowed for statistically significant correlations of inhibition results with coarse compound characteristics like clogP and molecular weight or volume. contact: Dr. Thomas Hesterkamp Evotec OAI AG [email protected] Schnackenburgallee 114 22525 Hamburg (Germany)

Stefan Welte, Günter Müller, Matthias Dreyer, Stefan Petry, Norbert Tennagels, Robert Schwenk Use of PTP1B mutants for selective inhibitor screening Protein-tyrosine phosphatase 1B (PTP 1B) has been implicated as a negative regulator of insulin action and as a potentially mediator in the pathogenesis of insulin-resistance and non-insulin dependent diabetes. Inhibiting this enzyme and preventing the insulin receptor downregulation has been suggested as a target for the treatment of type II diabetes. The development of highly selective Inhibitors for this enzyme requires direct insight into the structural elements that determine the substrate binding pocket. The 3D structure for more than 10% of the PTP family members has been solved. In the case of PTP1B substrate specificity is supposed to depend on recognition of the catalytic cleft and a secondary binding pocket. Altering this site by point mutation might be a way to identify selective small molecule inhibitors. Utilizing the crystal structure information of PTP1B we identified several amino acid residues as possible selectivity modules. The exchange of these residues alters the kinetic parameters and reduces the substrate recognition capacity using a peptide substrate derived from the insulin receptor. In contrast, no differences were observed when a small fluorogenic substrate was used indicating that the mutations did not alter the catalytic site. This was confirmed by detailed crystal structure analysis and correlated with the inhibitory properties of small molecules targeting either the catalytic cleft, the secondary binding site or both. Therefore the use of these mutants is a powerful tool to identify the binding region of inhibitors and will help to develop highly selective compounds. contact: Robert Schwenk Aventis Pharma Deutschland GmbH DG Metabolic Diseases [email protected] Industriepark Höchst 65926 Frankfurt am Main (Germany)

Andreas Bichet, Frank Hannemann, Rita Bernhardt Using the yeast two-hybrid system as in vivo screening-system for interactions between adrenodoxin and its redoxpartners Adrenodoxin (Adx) is a [2Fe - 2S] ferredoxin involved in steroid hormone biosynthesis in the adrenal gland mitochondrial matrix of mammals. It is a small soluble protein that transfers electrons from adrenodoxin reductase (AR) to different cytochrome P450 isoforms where they are consumed in hydroxylation reactions. The yeast two-hybrid system is a technique to detect interactions between two proteins. It was developed to provide a genetic means of identifying proteins that physically interact in vivo. We adapted this system to our purposes so that it can be used as a screening-system for adrenodoxin mutants and cytochrome P450 mutants to detect proteins with increased interaction to their respective partners. Different adrenodoxin mutants were generated by site-directed mutagenesis which lead to better interactions between adrenodoxin and its redox partners. On the one hand these results are important to complete the knowledge of the electron transfer from Adx to its redox partners by showing the importance of specified amino acid residues for this transfer. On the other hand there is a direct correlation between interaction and steroid production showing that the results obtained might be of relevance for biotechnological production of steroids. Literature Fields et al., Nature 1989; 340: 245-246 Frederickson, Current Opinion in Biotechnology 1998; 9:90-96 Schiffler et al., The Journal of Biological Chemistry 2001; 276: 36225-36232 contact: Diplom Biologe Andreas Bichet Universität des Saarlandes Biochemie Ak Prof.Bernhardt [email protected] im Stadtwald, Gebäude 9 Postfach 151150 66041 Saarbrücken (Deutschland)

Paul Pfluger, Marc Birringer, Regina Brigelius-Flohé, Ralph Rühl, Nico Landes vitamin E Vitamin E contact: Nico Landes Deutsches Institut f. Ernährungsforschung [email protected] Arthur Scheunert Allee 114-116 14558 Bergholz Rehbrücke (Brandenburg)

Peter Kühl Who discovered the Michaelis-Menten hyperbola? There exist a number of eponyms which carry the names of L. Michaelis (1875-1949) and M.L. Menten (1879-1960) and derive from their sole joint paper(1), e.g., Michaelis-Menten [MM] complex, MM constant, MM equation, MM hyperbola, MM hypothesis, MM mechanism, MM plot, MM theory. All these eponyms – with one exception, the MM constant Km – are incorrect (or of questionable legitimation) because they give M & M undeserved credit for the originality and priority of concepts which M & M never claimed for themselves; actually, M & M adopted expressis verbis the ideas of V. Henri(2) (1872-1940) and corroborated them by an improved experimental technique. Concerning the “MM hyperbola” it may also be mentioned that M & M(1) – contrary to previous enzyme kineticists(2,3) – neither used the term “hyperbola” or “hyperbolic” nor did they present their kinetic data as hyperbolic graphs. Clearly, the discovery of hyperbolic enzyme kinetics and its mass-action theoretical foundation belong to the achievements of Henri, not to those of M & M; for this reason, the usage of the misnomers “MM hyperbola” and “MM kinetics” should be discontinued. To eradicate or change an established eponym is difficult but not impossible as an example in physics illustrates: the unit of radioactivity was changed in 1975 from curie to becquerel in order to honour the true discoverer of radioactivity. So why not rename “MM kinetics” after the true discoverer and founder of hyperbolic kinetics into “Henri kinetics”? Literature 1) Michaelis, L., and Menten, M. L. (1913). Biochem. Z. 49, 333-369. 2) Henri, V. (1903). Lois générales de l’action des diastases (Paris: Librairie Scientifique A. Hermann). 3) Philoche, C. (1908). J. Chim. Phys. 6, 212-293 and 355-423. contact: Dr. Peter Kühl University of Basel Institute of Theoretical Biology [email protected] Schaulistr. 2 4142 Münchenstein BL (Switzerland)

Kerstin Reichert, Matthias Rödel, Ralph Menzel Xenobiotically induced cytochrome P450 gene expression in Caenorhabditis elegans The nematode Caenorhabditis elegans possesses the remarkable number of 80 different cytochrome P450 genes. The functions of the encoding proteins are almost unknown. In order to study xenobiotically induced gene expression in C. elegans liquid cultures were exposed to different well-known xenobiotic inducers. The mRNA expression was detected by two different DNA arrays and semi-quantitative RT-PCR (1). In addition, using concentration-effect experiments, a classical reproduction test was compared with CYP gene expression screens. Beta-naphthoflavone, PCB52, Lansoprazol and Fluoranthene were the most active CYP inducers. They mainly induced almost all CYP35 isoforms. The reproduction test with nine different xenobiotics revealed Tributylzinn, Endosulfan and Fluoranthene as most toxic substances. The threshold of the semi-quantitative RT-PCR experiments showed a significant higher sensitivity than the reproduction tests. Obviously, the xenobiotic inducible gene expression of C. elegans is a very sensitive tool to reveal defense mechanisms against potential toxic substances and can be used to develop a biomonitoring system (2). Interestingly, the CYP35 isoforms promoter regions contain xenobiotic response elements (XRE) similar to mammalian CYP1 forms. A transgenic C. elegans line expressing GFP under control of the CYP35A2 promoter was constructed. With this tool it was possible to follow the GFP production in vivo depending on the added inducer. In the control worms a slight but distinct label was visible at the anterior and posterior part of the intestine. In beta-naphthoflavone treated worms the total gut emitted a very strong GFP fluorescence. Literature (1) Menzel, R., Bogaert T. and Achazi A. (2001) A systematical gene expression screen of the Caenorhabditis elegans cytochrome P450 genes revealed CYP35 as strong xenobiotic inducible. Arch. Biochem. Biophys. 395, 158-168 (2) Menzel, R., Reichert, K. and Achazi A. (2002) Nutzung der induzierbaren Genexpression des Nematoden Caenorhabditis elegans als Biomonitior. UWSF 14, 18-23. contact: Dr. Ralph Menzel Free University Berlin Institute of Biology - Ecotoxicology & Biochemistry [email protected] Ehrenbergstr. 26-28 10319 Berlin (Germany)

Ross Dalbey YidC, a novel evolutionarily conserved protein, mediates membrane protein assembly in bacteria Membranes contain proteins that catalyze a variety of reactions, which lead to the selective permeability of the membrane. For membrane proteins to function as receptors, transporters, channels, and ATPases, they must be targeted to their correct membrane and inserted into the lipid bilayer. Our goal is to understand the biogenesis of polytopic membrane proteins in bacteria. While most E. coli membrane proteins use the Sec YEG pathway for insertion into the plasma membrane, there are some proteins that insert independent of the Sec pathway. Recently we identified a new membrane component called YidC that is essential for insertion of these Sec-independent proteins. YidC is essential for cell viability and is found in mitochondria and chloroplasts. Depletion of YidC interferes also with the insertion of Sec-dependent membrane proteins. We find that YidC directly interacts with membrane proteins during the process of membrane protein insertion. The chloroplast homolog Albino3 can functionally complement the bacterial YidC depletion strain, demonstrating that the chloropast and bacterial YidC homologs are truly functional homologs. The function of YidC for Sec-independent proteins may be analogous to a chaperone because YidC has been shown to bind and fold the inserting membrane proteins and not to interact with the fully synthesized and assembled protein. For sec-dependent proteins, YidC is proposed to catalyze the movement of hydrophobic regions out of the Sec channel and into the lipid bilayer.

contact: Professor Ross Dalbey Ohio State University Department of Chemistry [email protected] 100 West 18th Ave 43210 Columbus,OH (USA)

Authors (total 987) Name Abdelmohsen, Kotb Aigner, Achim Aktories, Klaus Aktories, Klaus Albig, Werner Alexander, Navarrete Santos Altmann, Reinhold Alves, Jürgen Ammon, Susanne Andrä, Jörg Arand, Michael Arbuckle, Julian Asakura, Atsushi Asperger, Otmar Asperger*, Otmar Augsten, Martin

Augustin, Robert

Aussel, Gudrun Baader, Stephan L. Baake, Matthias Bache, Matthias Bache, Matthias Baer, Patrick Baker, T. Ball, Claudia Baltensperger, Kurt Baltrusch, Simone Bamann, Christian Ban, Nenad Banning, Antje Bär, Jörg Baraka, A. M.

Abstract Activation of EGF receptor-dependent signaling pathways by alkylating and redox-cycling quinones Ribozyme-PEI-complexes as ready-to-use reagents for efficient down-regulation of gene expression Clostridium botulinum C3 exoezyme like ADP-ribosyltransferases Clostridium botulinum C2 toxin as a protein transport system Nuclear import of histones On the use of a histone-like protein for drug delivery using doxorubicin as a model system for tumor therapy Molecular modeling as a tool to identify putative interacting sites in the IL-1 receptor complex Restriction endonuclease EcoRI - fusions for a change of specificity Microarray analysis fo genes expressed in the frontal cortex of rats chronically treated with morphine Molecular basis for membrane selectivity of a potent peptide antibiotic derived from NK-lysin Structure-function relationships of epoxide hydrolases Pharmacogenetic based clinical trials to improve drug development Myogenic Specification of Adult Stem Cells in Skeletal Muscle P450non System from Acinetobacter sp. EB104: A Unique Bacterial Alkane Hydroxylase Sequence Analysis of an Acinetobacter Plasmid Carrying the Genes of a P450-Dependent Alkane Monooxygenase Multivalent Scavengers of Oncogenic Ras: A novel approach to tracking and therapeutical inhibition of Ras activity Induction of apoptotic cell death in bovine cumulus-oocyte complexes after treatment with Aroclor-1254 during in vitro maturation. PRO- AND ANTIAPOPTOTIC EFFECTS OF QUERCETIN AND FISETIN IN H4IIE-CELLS: IMPLICATION OF OXIDATIVE STRESS Tenascins are associated with lipid rafts isolated from mouse brain Nuclear import of histones Elevated expression level of survivin protein in soft-tissue sarcomas Growth reduction of a xenotransplanted human Soft Tissue Sarcoma by MDM2 Antisense Therapy Aldosterone action in humans -new members of the aldosterone signaling pathwayATP-dependent protein degradation and unfolding by ClpXP Analysis of oxidative stress due to fly ash and ultrafine particles in macrophages and epithelial cells of the lung SDF-1 and thrombin signaling in human hematopoietic progenitor cells is regulated by cytokines Effect of PFK-2 overexpression in insulin-producing cells upon glucokinase activity and glucose metabolism High-temperature structure of TmCsp Cytosolic proteins at birth: linking translation and chaperone assisted protein folding Redox events in IL-1 signaling Interactions between the two types of subunits forming an enzymatically active oligomeric phosphofructokinase-1 from Saccharomyces cerevisiae Effect of Lacidipine on induced liver fibrosis / cirrhosis.

Barbirz, Stefanie Barbosa, Eduardo Barbosa Sicard, Eduardo Bartel, Frank Bartel, Frank Barth, Gerold Barth, Henning Barth, Sandra Bartsch, Joerg W. Bartunik, Hans D. Bauer-Manz, Gabi Bäuerle, Marc Baumann, Frank Baumann, Ralf Bechert, Kim Beck, Mike Beck, Sabine Becker, Cord-Michael Behne, Dietrich Bengtsson, Luiza Bergmann, Jana Bergmann, Silke Bergmann, Silke Bernhagen, Jürgen Bernhardt, Rita Bernhardt, Rita Bernhardt, Rita

Berthold, F. Besong, Daniela Bichet, Andreas

Bier, Frank Bier, Frank F.

The tailspike from E.Coli H bacteriophage HK620: Another Parallel beta-Helix Protein? Cytochrome P450-dependent eicosapentaenoic acid epoxygenation produces novel vasoactive metabolites Metabolism of eicosapentaenoic acid by CYP4A and CYP2C enzymes Elevated expression level of survivin protein in soft-tissue sarcomas Growth reduction of a xenotransplanted human Soft Tissue Sarcoma by MDM2 Antisense Therapy FUNCTIONAL EXPRESSION OF HETEROLOGOUS CYTOCHROME P450 IN YARROWIA LIPOLYTICA AND ITS USE IN BIOTRANSFORMATION OF STEROIDS Aut10p, Mai1p and Aut5p shed new lights on the molecular mechanisms of autophagy in S.cerevisiae Polyomavirus-like particles as a system for gene therapy and drug delivery Tumor necrosis factor alpha signalling in reactive glia cells Crystal structure of the noncollagenous (NC1) domain of human placenta collagen IV: Stabilization via a novel type of covalent Met-Lys cross-link The E.coli membrane insertase YidC: A Chaperone for Protein Folding into the Membrane Nuclear import of histones Protein insertion into the inner membrane of mitochondria MACROPHAGE MIGRATION INHIBITORY FACTOR (MIF) IS A SURVIVAL FACTOR FOR NEUTROPHILS A new world of tiny RNAs - siRNAs and miRNAs GRIF-1 - a novel GABAA receptor associated protein Exogenous phospholipases A2 mediate gene expression in rat mesangial cells via PPARalpha activation Educational aspects of molecular medicine in basic science and clinical training Selenoproteins in the perinuclear structures of rat kidney Dedt. Molecular Trace Element research in the Life Sciences Hahn- Meitner-Istitut, Glienicker Str.100, 14109 Berlin Characterization of Inner Nuclear Membrane Proteins Nasal DNA vaccination with an Interleukin-10 cDNA vector in a murine asthma modell Analysis of EWS/Fli-1 induced gene expression in HEK293 cells using DNA-microarrays From genes to antigens: Potential target structures for autologous and allogeneic cytotoxic T cells identified by oligonucleotide arrays Investigations on the binding of SYBR Green I to double-stranded (ds)DNA Cytochrome P450 dependent steroid hydroxylases and their application in biotechnology and medicine Production of steroid hormones by fission yeast cells expressing human mitochondrial cytochromes P450 Using the yeast two-hybrid system as in vivo screening-system for interactions between adrenodoxin and its redoxpartners Classification and diagnostic prediction of pediatric cancers using gene expression profiling and artificial neural networks Genexpression of the human membrane-associated Progesterone Receptor (hmPR) in HepG2 cells Using the yeast two-hybrid system as in vivo screening-system for interactions between adrenodoxin and its redoxpartners On-chip synthesis of extended, long-chain DNA by PCR Xenia Marschan, Dennie Andresen, Markus von Nickisch-Rosenegk, Frank F. Bier Transcription of Reporter Genes with Immobilized Templates

Characterization of the human platelet-type 6-phosphofructo-1-kinase gene promoter Listeriolysin of Listeria monocytogenes forms Ca2+-permeable pores leading to intracellular Ca2+ Birringer, Jan oscillations in host cells vitamin E Birringer, Marc Expression of ubiquitous Bleichert*, Franziska 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase in intracranial neoplasms and non-tumorous brain tissue The intracellular N-terminus of aminopeptidase N/CD13 Blosz, Tanja affects association with caveolae Aminopeptidase N/CD13 is associated with Lubrol rafts Blosz, Tanja Crystal structure of the noncollagenous (NC1) domain of human placenta collagen IV: Stabilization via a novel Bode, Wolfram type of covalent Met-Lys cross-link Efficient Dosage and Time dependent Protein Transduction of Pancreatic Carcinoma Cells using purified Boenicke, Lars VP22-EGFP Fusion Protein Functional organization of the yeast proteome by Boesche, Markus systemic analysis of protein complexes Böhm, Christopher Interactionpartners of SorLA Polyomavirus-like particles as a system for gene therapy Böhm, Gerald and drug delivery Binding of RNA to the hyperthermophilic histone-like Böhm, Gerald protein TmHU INDUCTION OF THE HUMAN NEURONAL NITRIC OXIDE SYNTHASE EXPRESSION BY EPIDERMAL GROWTH Boissel, Jean-Paul FACTOR IS MEDIATED THROUGH THE JAK/STAT SIGNAL TRANSDUCTION PATHWAY CHARACTERIZATION OF THE MULTIPLE PROMOTERS OF Boissel, Jean-Paul THE HUMAN NEURONAL NITRIC OXIDE SYNTHASE GENE Redox events in IL-1 signaling Böl, Gaby Loss of function of the naturally occurring Arg219Leu Bönisch, Heinz variant of the human 5-HT1A receptor C(alpha)-Formylglycine: a novel protein modification in Borissenko, Ljudmila V. pro- and eukaryotes CYTOMEGALOVIRUS-A NEW VECTOR FOR GENE Borst, Eva TRANSFER Molecular modeling as a tool to identify putative Botzki, Alexander interacting sites in the IL-1 receptor complex Crystal structure of the noncollagenous (NC1) domain of human placenta collagen IV: Stabilization via a novel Bourenkov, Gleb P. type of covalent Met-Lys cross-link Oligomeric assembly of G protein-coupled receptors: Role Bouvier, Michel in receptor trafficking and function Brakmann, Susanne Optimal enzymes for single molecule sequencing Interaction of the organic base cimetidine with the renal Brändle, Edgar basolateral p-aminohippurate (PAH) transport system The TIM22 translocase mediates insertion of carrier precursors into the mitochondrial inner membrane via a Brandner, Katrin two-step process Breast Cancer and Predictive Factors: Association with Brauch, Hiltrud Polymorphisms Regulation of muscle cell development and regeneration Braun, Thomas by growth factors and bHLH proteins Evolution of Feedback-inhibited (&beta/&alpha)8 Barrel Braus, Gerhard Isoenzymes by Gene Duplication and A Single Mutation Trafficking pathways of connexin49-GFP in living Breidert, S. mammalian cells Natural ligand identification and functional Brézillon, Stéphane characterization of orphan G protein-coupled receptors GRIF-1 - a novel GABAA receptor associated protein Brickley, Kieran Brigelius-Flohé, Redox events in IL-1 signaling Regina Brigelius-Flohé, Regina vitamin E Listeriolysin of Listeria monocytogenes forms Ca2+-permeable pores leading to intracellular Ca2+ Brockmeier, Dierk oscillations in host cells Bigl, Marina

Brockmöller, Jürgen Brodde, Otto-Erich Bros, Matthias

Bross, Matthias

Bruck, Heike Brüning, Thomas Brunner, Eike Brunner, Herwig Brüss, Michael Bucci, Enrico Buchmann, Albrecht Bukau, Bernd Bukau, Bernd

Buniatian, Gayane²

Burdach, Stefan Burdach, Stefan Burdach, Stefan

Burdach, Stefan Burdach, Stefan Bureik, Matthias Burgering, Boudewijn

Pharmacogenetic studies on drug-metabolizing cytochromes P450 and possibilities for individual drug therapy Beta-adrenoceptor polymorphisms: does altered in vitro function predict in vivo effects? CHARACTERIZATION OF THE MULTIPLE PROMOTERS OF THE HUMAN NEURONAL NITRIC OXIDE SYNTHASE GENE INDUCTION OF THE HUMAN NEURONAL NITRIC OXIDE SYNTHASE EXPRESSION BY EPIDERMAL GROWTH FACTOR IS MEDIATED THROUGH THE JAK/STAT SIGNAL TRANSDUCTION PATHWAY Beta-adrenoceptor polymorphisms: does altered in vitro function predict in vivo effects? Breast Cancer and Predictive Factors: Association with Polymorphisms High-temperature structure of TmCsp Investigations on the binding of SYBR Green I to double-stranded (ds)DNA Loss of function of the naturally occurring Arg219Leu variant of the human 5-HT1A receptor Structure of an RNA Involved in Enteroviral Replication Promotion of hepatocarcinogenesis is mediated by overexpression of glutamine synthetase in beta-catenine-mutated mouse liver tumors Cytosolic proteins at birth: linking translation and chaperone assisted protein folding Structure-function analysis of HscC, the E. coli member of a novel subfamily of specialized Hsp70 chaperones Inverse Dynamics of Glial Fibrillary Acidic Protein (GFAP) Expression and Synthesis of Erythropoietin (EPO), EPO-Receptor (EPOR) and Proliferating Cell Nuclear Antigen (PCNA) in Astrocytes Analysis of EWS/Fli-1 induced gene expression in HEK293 cells using DNA-microarrays Nasal DNA vaccination with an Interleukin-10 cDNA vector in a murine asthma modell Differential analysis of interleukin 2 (IL-2) versus IL-15 induced immediate-early genes: A contribution for the elucidation of the AICD paradoxon? From genes to antigens: Potential target structures for autologous and allogeneic cytotoxic T cells identified by oligonucleotide arrays Cleavage of EWS/FLI-1 fusion transcripts by Hammerhead Ribozymes Production of steroid hormones by fission yeast cells expressing human mitochondrial cytochromes P450 cell cycle and death control: long live Forkheads

ATP-dependent protein degradation and unfolding by ClpXP NHE-1 inhibitors: From acute cardioprotection to heart Busch, Andreas failure Beta-adrenoceptor polymorphisms: does altered in vitro Büscher, Rainer function predict in vivo effects? Investigations on the binding of SYBR Green I to Buta, Christiane double-stranded (ds)DNA The knockout approach to understand the function of Buters, Jeroen xenobiotic metabolising enzymes Protein therapy: Applications in tissue regeneration and Cardoso, M. Cristina stem cell research Pharmacogenetics in the Cardiovascular Field Cascorbi, Ingolf Advanced Glycation Endproducts induced cell signalling in Casselmann, Christian cardiac fibroblasts Caspase inhibition strongly improves the recovery of cassens, uwe cryopreserved hematopoietic and other cells Listeriolysin of Listeria monocytogenes forms Ca2+-permeable pores leading to intracellular Ca2+ Chakraborty, Trinad oscillations in host cells PRO- AND ANTIAPOPTOTIC EFFECTS OF QUERCETIN AND FISETIN IN H4IIE-CELLS: IMPLICATION OF OXIDATIVE Chovolou, Yvonni STRESS Burton, R.

H2O2-INDUCED OXIDATIVE STRESS IN H4IIE CELLS: INFLUENCE OF THE FLAVONOID KAEMPFEROL ALTERED SENSITIVITY OF TNF-ALPHA OVEREXPRESSING RAT HEPATOMA CELLS AGAINST EXOGENOUS TNF-ALPHA Chovolou, Yvonni AND OXIDANTS On the use of a histone-like protein for drug delivery Claudia, Immisch using doxorubicin as a model system for tumor therapy On the use of a histone-like protein for drug delivery Claudia, Reinke using doxorubicin as a model system for tumor therapy Enzym polymorphisms and risk factors in bladder cancer Claudia, Roetzel patients in Lutherstadt Wittenberg Effect of BMP-2 on the migratory and invasive properties Clement, Joachim of breast cancer cells Natural ligand identification and functional Communi, David characterization of orphan G protein-coupled receptors Activities of oncogenic STAT3 in colorectal cancer cells Corvinus, Florian Proteins mediating vectorial transport across plasma Cui, Yunhai membrane domains Ribozyme-PEI-complexes as ready-to-use reagents for Czubayko, Frank efficient down-regulation of gene expression YidC, a novel evolutionarily conserved protein, mediates Dalbey, Ross membrane protein assembly in bacteria Tools of bioinformatics: Comparative genome analysis Dandekar, Thomas and pathway reconstruction Inverse Dynamics of Glial Fibrillary Acidic Protein (GFAP) Expression and Synthesis of Erythropoietin (EPO), Danielyan, Lusine¹ EPO-Receptor (EPOR) and Proliferating Cell Nuclear Antigen (PCNA) in Astrocytes Advanced Glycation Endproducts induced cell signalling in Daoud, Sherif cardiac fibroblasts Listeriolysin of Listeria monocytogenes forms Ca2+-permeable pores leading to intracellular Ca2+ Darji, Ayub oscillations in host cells Characterization of Inner Nuclear Membrane Proteins de Graaf, Daniel Therapeutic modulation of death receptor- or Debatin, Klaus-Michael drug-mediated apoptosis Characterization of Inner Nuclear Membrane Proteins Denner, Philip Protein therapy: Applications in tissue regeneration and Derer, Wolfgang stem cell research The human inositol phosphate multikinase (IPMK) is Deschermeier, targeted to the nucleus of mammalian cells by a basic Christina cluster in its C-terminus Natural ligand identification and functional Detheux, Michel characterization of orphan G protein-coupled receptors Involvement of different signalling pathways in cytoskeletal reorganisation during tributyltin (TBT) and Detzel, Tanja H2O2 induced apoptosis Cytosolic proteins at birth: linking translation and Deuerling, Elke chaperone assisted protein folding Use of cytochrome P450 isoenzyme assays in the Deul, Annette hit-to-lead process of drug discovery ß-amyloid peptide potentiates inflammatory responses induced by lipopolysaccharide, interferon-gamma and Deuther-Conrad, Winnie "advanced glycation endproducts" in a mouse microglia cell line Proinflammatory signal transduction pathways in N11 Deuther-Conrad, mouse microglial cell line activated by "Advanced Winnie Glycation Endproducts" Differential regulation of gap junction protein expression Dhein, Stefan in cardiomyocytes Effects of vitamin E on endothelial function in diabetes Dhein, Stefan mellitus and hyperglycemia Oxidative stress in rat alveolar epithelial cells and Diabaté, Silvia co-cultures upon treatment with fly ash Analysis of oxidative stress due to fly ash and ultrafine Diabate, Silvia particles in macrophages and epithelial cells of the lung C(alpha)-Formylglycine: a novel protein modification in Dierks, Thomas pro- and eukaryotes The role of NCAM phosphorylation on NCAM mediated Diestel, Simone signal transduction pathways Chovolou, Yvonni

Dietz, Frank Dihazi, Hassan Doenecke, Detlef Domann, Eugen Doods, Karin Dormann, Danuta Dove, Stefan Dragan, Calin-Aurel Dreißigacker, Ute Dreyer, Florian Dreyer, Matthias Dudeck, Ingrid Dudeck, Ingrid Dukic-Stefanovic, Sladjana Dukic-Stefanovic, Sladjana Dukic-Stefanovic, Sladjana Dürrschmidt, Peter Easwaran, Hariharan P. Eckhardt, Matthias Edelmann, Anke Ehrbar, Kristin Eichelbaum, Michel Eichelbaum, Michel El-Mallah ***, A. I. El-Reweni, S. H. El-Sharaki **, O. Elbashir, Sayda M. Elsner, Ralph-Peter Enenkel, Cordula Engeland, Kurt Epe, Bernd Epple, Ulrike D. Eschrich, Klaus

Characterization of a new member of the Hepatoma-derived growth factor protein family The Mechanism of regulation of yeast 6-phosphofructo-2-kinase by glucose and hypertonic stress Nuclear import of histones Listeriolysin of Listeria monocytogenes forms Ca2+-permeable pores leading to intracellular Ca2+ oscillations in host cells INHIBITION OF EROD-ACTIVITY IN RAT HEPATOMA CELLS Putative role of the nuclear factor NCNF in neuronal differentiation Molecular modeling as a tool to identify putative interacting sites in the IL-1 receptor complex Production of steroid hormones by fission yeast cells expressing human mitochondrial cytochromes P450 K-Ras-dependent signal tranduction in human pancreatic carcinoma cells Listeriolysin of Listeria monocytogenes forms Ca2+-permeable pores leading to intracellular Ca2+ oscillations in host cells Use of PTP1B mutants for selective inhibitor screening Targeting of chimaeric GFP-polypeptide fusions within chloroplasts Import and plastid sorting of chimaeric GFP-polypeptides Differential effects of "advanced glycation endproducts" and ß-amyloid peptide on glucose utilization and ATP levels in the neuronal cell line SH-SY5Y ß-amyloid peptide potentiates inflammatory responses induced by lipopolysaccharide, interferon-gamma and "advanced glycation endproducts" in a mouse microglia cell line Proinflammatory signal transduction pathways in N11 mouse microglial cell line activated by "Advanced Glycation Endproducts" Identification of an intermediate in the unfolding of neutral protease from Bacillus stearothermophilus by fluorescence spectroscopy Protein therapy: Applications in tissue regeneration and stem cell research Effect of N-glycan removal on catalytic activity of cerebroside sulfotransferase Interactions between the two types of subunits forming an enzymatically active oligomeric phosphofructokinase-1 from Saccharomyces cerevisiae Identification of the secretion and translocation domain of Salmonella typhimurium effector protein SopE FUNCTIONAL SIGNIFICANCE OF GENETIC POLYMORPHISMS OF HUMAN CYP2B6 ThioTEPA and Clopidogrel are specific mechanism-based inhibitors of human CYP2B6 Effect of Lacidipine on induced liver fibrosis / cirrhosis. Effect of Lacidipine on induced liver fibrosis / cirrhosis. Effect of Lacidipine on induced liver fibrosis / cirrhosis. A new world of tiny RNAs - siRNAs and miRNAs Solution structure of the Ras binding domain of AF6 Targeting mechanism of proteasomes to nuclear / ER membranes and into the nucleus of yeast Apoptosis induction and cell cycle regulation at the G2/M-checkpoint by p63, p73 and the tumor suppressor p53 Relevance of endogenous oxidative DNA damage: lessons from repair-deficient mice Aut10p, Mai1p and Aut5p shed new lights on the molecular mechanisms of autophagy in S.cerevisiae The Mechanism of regulation of yeast 6-phosphofructo-2-kinase by glucose and hypertonic stress

Nitrogen oxide-induced cell death in rabbit cultured Müller cells is modulated by glucose Characterization of the human platelet-type Eschrich, Klaus 6-phosphofructo-1-kinase gene promoter Expression of ubiquitous 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase in Eschrich*, Klaus intracranial neoplasms and non-tumorous brain tissue Refolding and Characterization of the N-terminal extracellular fragment of the Pituitary Adenylate-Cyclase Eugster, Marcel Activating Peptide Receptor Comparison of the effects of the P2Y12 ADP-receptor antagonist (AR-C69931) and the GPIIb/IIIa antagonist Eylenstein, Roy GR 144053F on platelet-leukocyte conjugate formation. Molecular modeling as a tool to identify putative Falk, Werner interacting sites in the IL-1 receptor complex C(alpha)-Formylglycine: a novel protein modification in Fang, Qinghua pro- and eukaryotes The human inositol phosphate multikinase (IPMK) is targeted to the nucleus of mammalian cells by a basic Fanick, Werner cluster in its C-terminus The role of NCAM phosphorylation on NCAM mediated Fergen, Marie-Therese signal transduction pathways Effect of N-glycan removal on catalytic activity of Fewou, Simon cerebroside sulfotransferase C(alpha)-Formylglycine: a novel protein modification in Fey, Jens pro- and eukaryotes Pharmacogenetic profiling with Biochips: an innovative Fiebich, Bernd concept to improve the efficiency of clinical trials Induction of apoptotic cell death in bovine cumulus-oocyte complexes after treatment with Fischer, Bernd Aroclor-1254 during in vitro maturation. The insulin receptor and the insulin-dependent glucose transporter isoform 4 are expressed in rabbit Fischer, Bernd preimplantation embryos Unfolding the Chemistry of Protein Folding Fischer, Gunter Breast Cancer and Predictive Factors: Association with Fischer, Hans-Peter Polymorphisms Fitzgerald, Garret A. The role of cyclooxygenases in the cardiovascular system Molecular tools to design trypanocidal drugs Flohé, Leopold The element of the moon moonlighting for fertility Flohé, Leopold Characterisation of Protein Domains Responsible for Florian, Volker Recognition of P-Selectin by Sorting Nexin 17 Isolation of an Arabidopsis T-DNA mutant for the Fluegge, Ulf-Ingo chloroplast protein import receptor atToc33 ATP-dependent protein degradation and unfolding by Flynn, J. ClpXP GLUTATHIONE S-TRANSFERASE GSTP1 POLYMORPHISMS IN DE NOVO AND THERAPY-INDUCED HEMATOLOGIC Fonatsch, Christa MALIGNANCIES FUNCTIONAL EXPRESSION OF HETEROLOGOUS CYTOCHROME P450 IN YARROWIA LIPOLYTICA AND ITS Förster, André USE IN BIOTRANSFORMATION OF STEROIDS INDUCTION OF THE HUMAN NEURONAL NITRIC OXIDE SYNTHASE EXPRESSION BY EPIDERMAL GROWTH Förstermann, Ulrich FACTOR IS MEDIATED THROUGH THE JAK/STAT SIGNAL TRANSDUCTION PATHWAY CHARACTERIZATION OF THE MULTIPLE PROMOTERS OF Förstermann, Ulrich THE HUMAN NEURONAL NITRIC OXIDE SYNTHASE GENE MRP expression and modulation of/by glutathione in Foth, Heidi human lung cells Enzym polymorphisms and risk factors in bladder cancer Frank, Geller patients in Lutherstadt Wittenberg Activities of CYP11B isoforms from different species. Frank, Hannemann Differential subcellular distribution, structure and functions of the 14-3-3 proteins of the unicellular green Frank, Ronald alga Chlamydomonas reinhardtii Quantitative Proteomics with Escherichia coli Franke, Gudrun Signal transduction by SorLA, a member of the Franke, Inga LDL-receptor family Eschrich, Klaus

Characterization of a new member of the Hepatoma-derived growth factor protein family Tenascins are associated with lipid rafts isolated from Franken, Sebastian mouse brain Natural ligand identification and functional Franssen, Jean-Denis characterization of orphan G protein-coupled receptors The INAD signaling complex of Drosophila photoreceptor: Recombinant Expression and Assembly in a Cell Culture Franz, Claudia System SIGNALING AND TRANSCRIPTIONAL CONTROL OF VISCERAL MUSCLE AND HEART DEVELOPMENT IN Frasch, Manfred DROSOPHILA Stability and Folding of Internalin B - A Biophysical Freiberg, Alexander Characterization Nitrogen oxide-induced cell death in rabbit cultured Müller Frenzel, Jochen cells is modulated by glucose A Receptor for Glycolipid Headgroups Mediates Frick, Wendelin Insulin-mimetic Signaling in Rat Adipocytes Multivalent Scavengers of Oncogenic Ras: A novel approach to tracking and therapeutical inhibition of Ras Friedrich, Karlheinz activity Activities of oncogenic STAT3 in colorectal cancer cells Friedrich, Karlheinz DMRT Genes and Sex Determination in the Fish Froschauer, Alexander Xiphophorous Membrane affinity and pore forming properties of proteins/peptides - Easy and fast by using solid Fuchs, Kathrin supported lipid bilayers. Reduced NO synthesis during halothane exposure in Fuhrmann, Margarita stimulated rat alveolar macrophages Therapeutic modulation of death receptor- or Fulda, Simone drug-mediated apoptosis The cardiac transcription factor Nkx2-5 regulates the funke-kaiser, heiko alternative promoters of the endothelin-converting enzyme-1 gene in H9c2 cardiomyoblasts Refolding from Fragments: The case of the proteolytically Fürst, Frank processed Pea Lectin Elevated expression level of survivin protein in soft-tissue Füssel, Susanne sarcomas Stress-signalling downstream to p38 MAPK: Essential role Gaestel, Matthias of MAPKAP kinase 2 (MK2) Optimization of intein based isotope labeling methods for Gail, Robert NMR investigations of proteins "Carbonyl stress” by methylglyoxal causes mitochondrial Gariba de Garcia, dysfunction, ATP depletion and NMDA receptor Susana overstimulation in neurons ß-amyloid peptide potentiates inflammatory responses induced by lipopolysaccharide, interferon-gamma and Gasic-Milenkovic, Jovana "advanced glycation endproducts" in a mouse microglia cell line Proinflammatory signal transduction pathways in N11 Gasic-Milenkovic, mouse microglial cell line activated by "Advanced Jovana Glycation Endproducts" K-Ras-dependent signal tranduction in human pancreatic Gaugler, Monika carcinoma cells Characterization of the human platelet-type Gaunitz, Frank 6-phosphofructo-1-kinase gene promoter Promotion of hepatocarcinogenesis is mediated by overexpression of glutamine synthetase in Gaunitz, Frank beta-catenine-mutated mouse liver tumors Novel signalling pathways in striated muscles Gautel, Mathias Chaperons at the Ribosomal Tunnel Exit - First Contacts Gautschi, Matthias of a Newly Synthesized Polypeptide Functional organization of the yeast proteome by Gavin, Anne-Claude systemic analysis of protein complexes Promotion of hepatocarcinogenesis is mediated by overexpression of glutamine synthetase in Gebhardt, Rolf beta-catenine-mutated mouse liver tumors Modulation of spinal nociceptive processing through the Geisslinger, Gerd glutamate transporter GLT-1 Franken, Sebastian

A mathematical model to predict plasma concentrations of morphine and its active metabolite morphine-6-glucuronide in healthy young persons Activation of c-Jun-N-terminal-kinase by R- and S-flurbiprofen results in cell cycle arrest in human colon Geisslinger, Gerd carcinoma cells CyGene: Eine neue hefebasierte Methode zur Bestimmung des cyto- und genotoxischen Potentials von Gellert, Georg Umweltgiften im Wasserbereich. CyGene: A New Yeast Based Method for the Detection of Cyto- and Genotox On the use of a histone-like protein for drug delivery Gerald, Böhm using doxorubicin as a model system for tumor therapy FUNCTIONAL EXPRESSION OF HETEROLOGOUS CYTOCHROME P450 IN YARROWIA LIPOLYTICA AND ITS Gerber, Jana USE IN BIOTRANSFORMATION OF STEROIDS Pharmacogenetic studies on drug-metabolizing cytochromes P450 and possibilities for individual drug Gerloff, Thomas therapy Effect of BMP-2 on the migratory and invasive properties Geyer, Annette of breast cancer cells Ghoneim, M Tharwat Effect of Lacidipine on induced liver fibrosis / cirrhosis. K-Ras-dependent signal tranduction in human pancreatic Giehl, Klaudia carcinoma cells K-Ras-dependent signal tranduction in human pancreatic Gierschik, Peter carcinoma cells Characterization of a new member of the Gieselmann, Volkmar Hepatoma-derived growth factor protein family Tenascins are associated with lipid rafts isolated from Gieselmann, Volkmar mouse brain Effect of N-glycan removal on catalytic activity of Gieselmann, Volkmar cerebroside sulfotransferase The evolution of RNase P in eukaryotic organelles Gimple, Olaf Identification of aminocoumarin antibiotics from different Gleiter, Christoph Streptomyces species via ESI-MS and LC-MS/MS coupling Inverse Dynamics of Glial Fibrillary Acidic Protein (GFAP) Expression and Synthesis of Erythropoietin (EPO), Gleiter, Christoph¹ EPO-Receptor (EPOR) and Proliferating Cell Nuclear Antigen (PCNA) in Astrocytes INDUCTION OF THE HUMAN NEURONAL NITRIC OXIDE SYNTHASE EXPRESSION BY EPIDERMAL GROWTH Gödtel-Armbrust, Ute FACTOR IS MEDIATED THROUGH THE JAK/STAT SIGNAL TRANSDUCTION PATHWAY CHARACTERIZATION OF THE MULTIPLE PROMOTERS OF Gödtel-Armbrust, Ute THE HUMAN NEURONAL NITRIC OXIDE SYNTHASE GENE Comparison of the effects of the P2Y12 ADP-receptor antagonist (AR-C69931) and the GPIIb/IIIa antagonist Göhring, Isabel GR 144053F on platelet-leukocyte conjugate formation. Enzym polymorphisms and risk factors in bladder cancer Golka, Klaus patients in Lutherstadt Wittenberg Cytochrome P450-dependent eicosapentaenoic acid Gollasch, Maik epoxygenation produces novel vasoactive metabolites Structure of an RNA Involved in Enteroviral Replication Görlach, Matthias Loss of function of the naturally occurring Arg219Leu Göthert, Manfred variant of the human 5-HT1A receptor Nuclear receptors and their role in the regulation of Göttlicher, Martin xenobiotic metabolism Restriction endonuclease EcoRI - fusions for a change of Grabowski, Gabriele specificity Selenoproteins in the perinuclear structures of rat kidney Dedt. Molecular Trace Element research in the Life Graebert, Alexandra Sciences Hahn- Meitner-Istitut, Glienicker Str.100, 14109 Berlin Functional organization of the yeast proteome by Grandi, Paola systemic analysis of protein complexes ATP-dependent protein degradation and unfolding by Grant, R. ClpXP Comparison of the effects of the P2Y12 ADP-receptor antagonist (AR-C69931) and the GPIIb/IIIa antagonist Grau, Marion GR 144053F on platelet-leukocyte conjugate formation. Expression, Purification and Characterization of the Inner Grauschopf, Ulla Membrane Redox Catalyst DsbB from E. coli Geisslinger, Gerd

Refolding and Characterization of the N-terminal extracellular fragment of the Pituitary Adenylate-Cyclase Activating Peptide Receptor Interaction of the organic base cimetidine with the renal Greven, Joachim basolateral p-aminohippurate (PAH) transport system GLUTATHIONE S-TRANSFERASE GSTP1 POLYMORPHISMS IN DE NOVO AND THERAPY-INDUCED HEMATOLOGIC Griesinger, Frank MALIGNANCIES Activation of c-Jun-N-terminal-kinase by R- and S-flurbiprofen results in cell cycle arrest in human colon Groesch, Sabine carcinoma cells PHYSICAL INTERACTION BETWEEN NUCLEAR DNA HELICASE II AND WRN HELICASE STIMULATES WRN´S Grosse, Frank EXONUCLEASE ACTIVITY p53 is involved in the repair of human topoisomerase I Grosse, Frank cleavage complexes Mechanisms of Oxidations of Drugs and Carcinogens Guengerich, F. Peter Catalyzed by Human P450s Polyomavirus-like particles as a system for gene therapy Günther, Constanze and drug delivery P450non System from Acinetobacter sp. EB104: A Unique Gupta, Nishith Bacterial Alkane Hydroxylase Isolation of an Arabidopsis T-DNA mutant for the Gutensohn, Michael chloroplast protein import receptor atToc33 Comparison of the effects of the P2Y12 ADP-receptor antagonist (AR-C69931) and the GPIIb/IIIa antagonist Güttich, Sven GR 144053F on platelet-leukocyte conjugate formation. Restriction endonuclease EcoRI - fusions for a change of Ha-Thi, Minh-Cam specificity GLUTATHIONE S-TRANSFERASE GSTP1 POLYMORPHISMS IN DE NOVO AND THERAPY-INDUCED HEMATOLOGIC Haase, Detlef MALIGNANCIES Differential regulation of gap junction protein expression Hagendorff, Andreas in cardiomyocytes Sequence Analysis of an Acinetobacter Plasmid Carrying Hahn*, Ulrich the Genes of a P450-Dependent Alkane Monooxygenase Cytochrome P450-dependent eicosapentaenoic acid Haller, Hermann epoxygenation produces novel vasoactive metabolites GLUTATHIONE S-TRANSFERASE GSTP1 POLYMORPHISMS IN DE NOVO AND THERAPY-INDUCED HEMATOLOGIC Hallier, Ernst MALIGNANCIES Breast Cancer and Predictive Factors: Association with Hamann, Ute Polymorphisms Effect of Lacidipine on induced liver fibrosis / cirrhosis. Hammadi, S. H. Interactionpartners of SorLA Hampe, Wolfgang Signal transduction by SorLA, a member of the Hampe, Wolfgang LDL-receptor family A Receptor for Glycolipid Headgroups Mediates Hanekop, Nils Insulin-mimetic Signaling in Rat Adipocytes DMRT Genes and Sex Determination in the Fish Hanel, Reinhold Xiphophorous Fanconi anemia: basic molecular biology and clinical Hanenberg, Helmut implications Differential analysis of interleukin 2 (IL-2) versus IL-15 induced immediate-early genes: A contribution for the Hanewinkel, Karin elucidation of the AICD paradoxon? Characterization of the human platelet-type Hannemann, Anke 6-phosphofructo-1-kinase gene promoter Using the yeast two-hybrid system as in vivo screening-system for interactions between adrenodoxin Hannemann, Frank and its redoxpartners Hunting for proteins associated with the delta pH / Hanner, Peter Tat-pathway in chloroplasts Nasal DNA vaccination with an Interleukin-10 cDNA Hansen, Gesine vector in a murine asthma modell A new world of tiny RNAs - siRNAs and miRNAs Harborth, Jens Identification of the secretion and translocation domain of Hardt, Wolf-Dietrich Salmonella typhimurium effector protein SopE Cellular transformation mediated by Src kinase: the role Häring, Hans-Ulrich of PTPalpha Grauschopf, Ulla

Inverse Dynamics of Glial Fibrillary Acidic Protein (GFAP) Expression and Synthesis of Erythropoietin (EPO), Harsch, Simone¹ EPO-Receptor (EPOR) and Proliferating Cell Nuclear Antigen (PCNA) in Astrocytes Breast Cancer and Predictive Factors: Association with Harth, Volker Polymorphisms p53 is involved in the repair of human topoisomerase I Hartmann, Hella cleavage complexes Evolution of Feedback-inhibited (&beta/&alpha)8 Barrel Hartmann, Markus Isoenzymes by Gene Duplication and A Single Mutation G-CSF in infection prophylaxis Hartung, Thomas Interaction of the organic base cimetidine with the renal Haßler, Martin basolateral p-aminohippurate (PAH) transport system Phospholipase D from cabbage: Calcium ions essential for Haufe, Susanne activity destabilize the protein conformation Anti-infective intervention via Toll-like receptors Heeg, Klaus Characterizing the signaling pathway of rat p18 amyloid Heese, Klaus beta (Aß) responsive protein (p18AßrP). Identification of aminocoumarin antibiotics from different Heide, Lutz Streptomyces species via ESI-MS and LC-MS/MS coupling Enzym polymorphisms and risk factors in bladder cancer Heidi, Foth patients in Lutherstadt Wittenberg Temperature-sensitive TMV Coat Proteins as Pathogens in Heimann, Peter Animal Cells Evolution of Feedback-inhibited (&beta/&alpha)8 Barrel Heinrich, Gabriele Isoenzymes by Gene Duplication and A Single Mutation Folding Kinetics of Pectate Lyase from Bacillus subtilis Heinz, Benjamin Stability and Folding of Internalin B - A Biophysical Heinz, Dirk Characterization OPTIMIZED ANTISENSE OLIGONUCLEOTIDE-MEDIATED INHIBITION OF THE INDUCIBLE NITRIC OXIDE Hemmrich, Karsten SYNTHASE CO-SUPPRESSES NO-MEDIATED STRESS RESPONSE IN ENDOTHELIUM Crystal structure of the noncollagenous (NC1) domain of human placenta collagen IV: Stabilization via a novel Henrich, Stefan type of covalent Met-Lys cross-link Comparison of the effects of the P2Y12 ADP-receptor antagonist (AR-C69931) and the GPIIb/IIIa antagonist Heptinstall, Stan GR 144053F on platelet-leukocyte conjugate formation. Identification of the Major Tyrosin Phosphorylation Sites Herkner, Armin of Human Gab-1 Enzym polymorphisms and risk factors in bladder cancer Hermann M, Bolt patients in Lutherstadt Wittenberg Protein insertion into the inner membrane of Herrmann, Johannes mitochondria Analysis of Posttranslational Modifications of Herrmann, Marion alpha-A-Crystallin during Aging of the Eye Lens Catechins with a galloyl group in the 3-position inhibit the PDGF-BB-induced intracellular signal transduction Hescheler, Jürgen pathway and proliferation of vascular smooth muscle cells PDGF-BB induces selective differentiation of mouse Hescheler, Jürgen embryonic stem cells to cardiac cells Use of cytochrome P450 isoenzyme assays in the Hesterkamp, Thomas hit-to-lead process of drug discovery The evolution of RNase P in eukaryotic organelles Heubeck, Christian GLUTATHIONE S-TRANSFERASE GSTP1 POLYMORPHISMS IN DE NOVO AND THERAPY-INDUCED HEMATOLOGIC Heutelbeck, Astrid MALIGNANCIES Functions of the proteasome in cell cycle and apoptosis Hilt, Wolfgang Microarray analysis of mucosal gene expression profiles Höcker, Michael during gastric H. pylori infection Effect of BMP-2 on the migratory and invasive properties Höffken, Klaus of breast cancer cells FUNCTIONAL SIGNIFICANCE OF GENETIC Hofmann, Ute POLYMORPHISMS OF HUMAN CYP2B6 Optimization of intein based isotope labeling methods for Hofweber, Roland NMR investigations of proteins Clostridium botulinum C2 toxin as a protein transport Holger, Barth system

Holger, Dietrich Höllt, Volker Holmgren, Arne Homey, Bernhard Honeck, Horst Honeck, Horst Honeck, Horst

Hoppe, Barbara

Horn, Gudrun Hortschansky, Peter Hou, Bo Huber, Armin

Huber, Robert Hühn, Regina Humeny, Andreas Hust, Bianca Hutter, Christoph Hutter, Christoph Ilyina, Julia Immisch, Claudia Izumo, Seigo Jacob, R. Jäger, Christiane Jakob, Mario Jockusch, Harald Joshi, S. Jung, Astrid Jung, Heinrich Jungmann, Joern Juretzek, Thomas Just, Sören Justenhoven, Christina Jüttner, Stefan

Enzym polymorphisms and risk factors in bladder cancer patients in Lutherstadt Wittenberg Microarray analysis fo genes expressed in the frontal cortex of rats chronically treated with morphine Redox regulation by thioredoxin and glutaredoxin systems A role for Chemokine Receptors in Cancer Metastasis? Cytochrome P450-dependent eicosapentaenoic acid epoxygenation produces novel vasoactive metabolites Metabolism of eicosapentaenoic acid by CYP4A and CYP2C enzymes Cytochrome P450-Dependent Renal Metabolism of Arachidonic Acid in a Rat Model of Angiotensin II induced Hypertension and End-Organ Damage Selenoproteins in the perinuclear structures of rat kidney Dedt. Molecular Trace Element research in the Life Sciences Hahn- Meitner-Istitut, Glienicker Str.100, 14109 Berlin Optimization of intein based isotope labeling methods for NMR investigations of proteins Effect of BMP-2 on the migratory and invasive properties of breast cancer cells The mechanism of deltapH/TAT-dependent protein translocation across the thylakoid membrane The INAD signaling complex of Drosophila photoreceptor: Recombinant Expression and Assembly in a Cell Culture System Crystal structure of the noncollagenous (NC1) domain of human placenta collagen IV: Stabilization via a novel type of covalent Met-Lys cross-link Cleavage of EWS/FLI-1 fusion transcripts by Hammerhead Ribozymes MALDI-TOF-MS Based Analysis of Disease-related Gene Variants Isolation of an Arabidopsis T-DNA mutant for the chloroplast protein import receptor atToc33 Analysis of EWS/Fli-1 induced gene expression in HEK293 cells using DNA-microarrays From genes to antigens: Potential target structures for autologous and allogeneic cytotoxic T cells identified by oligonucleotide arrays Functions of the proteasome in cell cycle and apoptosis Binding of RNA to the hyperthermophilic histone-like protein TmHU The role of Csx/Nkx 2.5 homeobox gene in cardiac development and congenital heart disease. Trafficking pathways of connexin49-GFP in living mammalian cells Polyomavirus-like particles as a system for gene therapy and drug delivery Hunting for proteins associated with the delta pH / Tat-pathway in chloroplasts Temperature-sensitive TMV Coat Proteins as Pathogens in Animal Cells ATP-dependent protein degradation and unfolding by ClpXP High-temperature structure of TmCsp Targeting of proteins to the flagellar type III secretion system of Escherichia coli Use of cytochrome P450 isoenzyme assays in the hit-to-lead process of drug discovery FUNCTIONAL EXPRESSION OF HETEROLOGOUS CYTOCHROME P450 IN YARROWIA LIPOLYTICA AND ITS USE IN BIOTRANSFORMATION OF STEROIDS Chaperons at the Ribosomal Tunnel Exit - First Contacts of a Newly Synthesized Polypeptide Breast Cancer and Predictive Factors: Association with Polymorphisms Microarray analysis of mucosal gene expression profiles during gastric H. pylori infection

Effects of vitamin E on endothelial function in diabetes mellitus and hyperglycemia Cytochrome P450-dependent eicosapentaenoic acid Kaergel, Eva epoxygenation produces novel vasoactive metabolites PRO- AND ANTIAPOPTOTIC EFFECTS OF QUERCETIN AND FISETIN IN H4IIE-CELLS: IMPLICATION OF OXIDATIVE Kahl, Regine STRESS H2O2-INDUCED OXIDATIVE STRESS IN H4IIE CELLS: Kahl, Regine INFLUENCE OF THE FLAVONOID KAEMPFEROL ALTERED SENSITIVITY OF TNF-ALPHA OVEREXPRESSING RAT HEPATOMA CELLS AGAINST EXOGENOUS TNF-ALPHA Kahl, Regine AND OXIDANTS Nuclear import of histones Kahle, Jörg Identification of aminocoumarin antibiotics from different Kahlich, Rainer Streptomyces species via ESI-MS and LC-MS/MS coupling The endosomal/lysosomal distribution of cathepsins (cat) B, L and S and N-acetyl-ß-D-glucosaminidase (NAG) is Kalbacher, Hubert differentially modulated in murine cell lines of microglial and monocytic origin Optimization of intein based isotope labeling methods for Kalbitzer, Hans Robert NMR investigations of proteins Kalbitzer, Hans-Robert Are GTP-analogs really good analogs? Kalbitzer, Hans-Robert High-temperature structure of TmCsp Efficient Dosage and Time dependent Protein Transduction of Pancreatic Carcinoma Cells using purified Kalthoff, Holger VP22-EGFP Fusion Protein Identification of aminocoumarin antibiotics from different Kammerer, Bernd Streptomyces species via ESI-MS and LC-MS/MS coupling PRO- AND ANTIAPOPTOTIC EFFECTS OF QUERCETIN AND Kampkötter, Andreas FISETIN IN H4IIE-CELLS: IMPLICATION OF OXIDATIVE STRESS H2O2-INDUCED OXIDATIVE STRESS IN H4IIE CELLS: Kampkötter, Andreas INFLUENCE OF THE FLAVONOID KAEMPFEROL ALTERED SENSITIVITY OF TNF-ALPHA OVEREXPRESSING Kampkötter, Andreas RAT HEPATOMA CELLS AGAINST EXOGENOUS TNF-ALPHA AND OXIDANTS GAF-domains as novel cAMP-sensor autoactivate a Kanacher, Tobias bacterial adenylyl cyclase Efficient Dosage and Time dependent Protein Transduction of Pancreatic Carcinoma Cells using purified Kang, Chu VP22-EGFP Fusion Protein Cellular transformation mediated by Src kinase: the role Kapp, Katja of PTPalpha Characterization of a new member of the Kappler, Joachim Hepatoma-derived growth factor protein family Tenascins are associated with lipid rafts isolated from Kappler, Joachim mouse brain Elevated expression level of survivin protein in soft-tissue Kappler, Matthias sarcomas Growth reduction of a xenotransplanted human Soft Kappler, Matthias Tissue Sarcoma by MDM2 Antisense Therapy Cytochrome P450-Dependent Renal Metabolism of Arachidonic Acid in a Rat Model of Angiotensin II induced Kärgel, Eva Hypertension and End-Organ Damage Functions of the proteasome in cell cycle and apoptosis Karnam, Harish Exogenous phospholipases A2 mediate gene expression in Kaszkin, Marietta rat mesangial cells via PPARalpha activation Effect of Lacidipine on induced liver fibrosis / cirrhosis. Kato *, S. Study on the function and kinetics of Katsemi, Vicky diisopropylfluorophosphatase Pharmacogenetic profiling with Biochips: an innovative Kaufmann, Martina concept to improve the efficiency of clinical trials Activities of oncogenic STAT3 in colorectal cancer cells Kaufmann, Roland THE EXPRESSION OF ANGIOGENIC GROWTH FACTORS IN HUMAN GRANULOSA CELLS ARE CONTROLLED BY HUMAN KECK, Christoph CHORION GONADOTROPIN. EXPRESSION OF A NEW SPLICE VARIANT OF IGFBP-7/MAC25 TUMOR SUPPRESSOR GENE IN HUMAN KECK, Christoph GRANULOSA CELLS LACKING THE IGFBP MOTIF (GCGCCXXC) Kabat, Armin

Kellersmann, Julia Kellner, Markus Kenniston, J. Keppler, Dietrich Keßler*, Renate Kettrup, Antonius Khan, J.

Kirberg, Bettina

Kirchberger, Jürgen

Kirchheiner, Julia Kirchhoff, Christian Kirschner, Karin Kirstein, Michaela Klein, Kathrin Klein, Kathrin Kloesgen, Ralf Bernd Kloesgen, Ralf Bernd Kloor1,

Doris

Klose, Joachim Klösgen, Ralf Bernd Klösgen, Ralf Bernd Klösgen, Ralf Bernd Klösgen, Ralf Bernd Klotz, Lars-Oliver Klotz, Lars-Oliver Klotz, Lars-Oliver Klotz, Lars-Oliver Kluck, Christoph Knauth, Peter Ko, Yon

The O-GlcNAc modification of proteins interferes with signaling by PKA and CDK5 dependent phosphorylation in neurons Aldosterone action in humans -new members of the aldosterone signaling pathwayATP-dependent protein degradation and unfolding by ClpXP Proteins mediating vectorial transport across plasma membrane domains Expression of ubiquitous 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase in intracranial neoplasms and non-tumorous brain tissue INHIBITION OF EROD-ACTIVITY IN RAT HEPATOMA CELLS Classification and diagnostic prediction of pediatric cancers using gene expression profiling and artificial neural networks CyGene: Eine neue hefebasierte Methode zur Bestimmung des cyto- und genotoxischen Potentials von Umweltgiften im Wasserbereich. CyGene: A New Yeast Based Method for the Detection of Cyto- and Genotox Interactions between the two types of subunits forming an enzymatically active oligomeric phosphofructokinase-1 from Saccharomyces cerevisiae Pharmacogenetic studies on drug-metabolizing cytochromes P450 and possibilities for individual drug therapy Use of cytochrome P450 isoenzyme assays in the hit-to-lead process of drug discovery SDF-1 and thrombin signaling in human hematopoietic progenitor cells is regulated by cytokines The insulin receptor and the insulin-dependent glucose transporter isoform 4 are expressed in rabbit preimplantation embryos FUNCTIONAL SIGNIFICANCE OF GENETIC POLYMORPHISMS OF HUMAN CYP2B6 ThioTEPA and Clopidogrel are specific mechanism-based inhibitors of human CYP2B6 Import and plastid sorting of chimaeric GFP-polypeptides Isolation of an Arabidopsis T-DNA mutant for the chloroplast protein import receptor atToc33 The ratio of the cofactor NAD +/NADH of S-adenosylhomocysteine hydrolase controls the adenosine binding sites. Analysis of Posttranslational Modifications of alpha-A-Crystallin during Aging of the Eye Lens Transport and assembly of the Rieske Fe/S protein: What happens in the stromal space? Targeting of chimaeric GFP-polypeptide fusions within chloroplasts Hunting for proteins associated with the delta pH / Tat-pathway in chloroplasts The mechanism of deltapH/TAT-dependent protein translocation across the thylakoid membrane (-)-Epicatechin Is Taken up by Cells and Protects against Peroxynitrite-Induced Oxidation and Nitration. Activation of EGF receptor-dependent signaling pathways by alkylating and redox-cycling quinones Oxidant-induced signaling: from intra- to intercellular communication Singlet Molecular Oxygen Reversibly Inhibits Tyrosine Phosphatases Structure-function analysis of HscC, the E. coli member of a novel subfamily of specialized Hsp70 chaperones Characterisation of Protein Domains Responsible for Recognition of P-Selectin by Sorting Nexin 17 Catechins with a galloyl group in the 3-position inhibit the PDGF-BB-induced intracellular signal transduction pathway and proliferation of vascular smooth muscle cells

Breast Cancer and Predictive Factors: Association with Polymorphisms SH2-domain containing inositol 5-phosphatase (SHIP)-1 is a potential regulator of cytoskeletal rearrangements in Koch, Alexandra NGF induced neurite outgrowth PHYSICAL INTERACTION BETWEEN NUCLEAR DNA HELICASE II AND WRN HELICASE STIMULATES WRN´S Köhler, Carsten EXONUCLEASE ACTIVITY Trafficking pathways of connexin49-GFP in living Kolb, H.-A. mammalian cells OPTIMIZED ANTISENSE OLIGONUCLEOTIDE-MEDIATED INHIBITION OF THE INDUCIBLE NITRIC OXIDE Kolb-Bachofen, Victoria SYNTHASE CO-SUPPRESSES NO-MEDIATED STRESS RESPONSE IN ENDOTHELIUM Genome wide search for disease genes as new drug Koller, Klaus-Peter targets Isolation of an Arabidopsis T-DNA mutant for the Kolukisaoglu, Uener chloroplast protein import receptor atToc33 Proteins mediating vectorial transport across plasma König, Jörg membrane domains Structural requirements for calmodulin binding to Konrad, Manfred membrane-associated guaylate kinases A designed variant of Herpes virus thymidine kinase selectively phosphorylates the anticancer and antiviral Konrad, Manfred prodrug ganciclovir Structural and functional analysis of deoxyribonucleoside kinases dCK and dGK: design of novel mutant enzymes Konrad, Manfred for therapeutic applications. cell cycle and death control: long live Forkheads Kops, Geert Polyomavirus-like particles as a system for gene therapy Körber, Sabine and drug delivery From genes to antigens: Potential target structures for autologous and allogeneic cytotoxic T cells identified by Körber, Sabine oligonucleotide arrays DMRT Genes and Sex Determination in the Fish Körting, Cornelia Xiphophorous Listeriolysin of Listeria monocytogenes forms Ca2+-permeable pores leading to intracellular Ca2+ Koschinski, Andreas oscillations in host cells Loss of function of the naturally occurring Arg219Leu Kostanian, Arina variant of the human 5-HT1A receptor Natural ligand identification and functional Kotani, Masato characterization of orphan G protein-coupled receptors Hsp90 effects on the regulation of soluble guanylyl Kots, Alexander Y. cyclase Elevated expression level of survivin protein in soft-tissue Kotzsch, Matthias sarcomas Effect of MAP kinase inhibitors on injury-induced Kovacevic, Slobodan neointima formation in the rat carotid artery The cardiac transcription factor Nkx2-5 regulates the Kovacevic, slobodan d. alternative promoters of the endothelin-converting enzyme-1 gene in H9c2 cardiomyoblasts The TIM22 translocase mediates insertion of carrier precursors into the mitochondrial inner membrane via a Kovermann, Peter two-step process Putative role of the nuclear factor NCNF in neuronal Koziollek-Drechsler, Ingrid differentiation Cytosolic proteins at birth: linking translation and Kramer, Günter chaperone assisted protein folding Functional organization of the yeast proteome by Krause, Roland systemic analysis of protein complexes Pharmacogenetics in the Cardiovascular Field Kroemer, Heyo K. Effect of BMP-2 on the migratory and invasive properties Kroll, Torsten of breast cancer cells High throughput gene expression profiling to screen for Kroll, Werner drug effects and toxicity Caspase-10 is the Key Initiator-Caspase involved in Krug, Harald F. TBT-mediated Apoptosis Oxidative stress in rat alveolar epithelial cells and Krug, Harald F. co-cultures upon treatment with fly ash Ko, Yon

Krug, Harald F. Krug, Harald F. Krug, Harald, F.

Krügel, V.

Kuçi, Selim³

Kühbacher, Marcus Kühl, Peter Kuhla, Björn

Kuhlmann, Juergen Kuhlmann, Jürgen Kuhn, Andreas Kühn, Diana

Kühn, Klaus Kuklinski, Stefan Kupper, Dagmar Kuster, Bernhard Kyriakopoulos, Antonios Lagos-Quintana, Mariana Lambeau, Gerard Lammers, Reiner Lämmle, Katrin Landes, Nico Lang, Thomas Langer, Thomas Langner, Juergen Langner, Jürgen Laufs, Ulrich Lautenschläger, Chritine Lauterbach, Birgit Le Poul, Emmanuel Lee, Hsiu-Hsiang Lehmann, Katrin

Analysis of oxidative stress due to fly ash and ultrafine particles in macrophages and epithelial cells of the lung Alkylphosphocholine Analogues induce Apoptosis in Tumour Cells of the Immune System Involvement of different signalling pathways in cytoskeletal reorganisation during tributyltin (TBT) and H2O2 induced apoptosis Inhibition of LiCl-induced induction of glutamine synthetase in HepG2 cells by flavonoids and anthraquinone derivatives is mediated by CK I Inverse Dynamics of Glial Fibrillary Acidic Protein (GFAP) Expression and Synthesis of Erythropoietin (EPO), EPO-Receptor (EPOR) and Proliferating Cell Nuclear Antigen (PCNA) in Astrocytes Selenoproteins in the perinuclear structures of rat kidney Dedt. Molecular Trace Element research in the Life Sciences Hahn- Meitner-Istitut, Glienicker Str.100, 14109 Berlin Who discovered the Michaelis-Menten hyperbola? Differential effects of "advanced glycation endproducts" and ß-amyloid peptide on glucose utilization and ATP levels in the neuronal cell line SH-SY5Y Development of an in vitro system for detection and quantitation of urothelial genotoxicity of tobacco smoke-specific constituents Localisation of fluorescently labelled Ras protein in the living cell The E.coli membrane insertase YidC: A Chaperone for Protein Folding into the Membrane Membrane affinity and pore forming properties of proteins/peptides - Easy and fast by using solid supported lipid bilayers. Crystal structure of the noncollagenous (NC1) domain of human placenta collagen IV: Stabilization via a novel type of covalent Met-Lys cross-link Altered O-GlcNAc level after neuronal differentiation of PC12 cells Redox events in IL-1 signaling Functional organization of the yeast proteome by systemic analysis of protein complexes Selenoproteins in the perinuclear structures of rat kidney Dedt. Molecular Trace Element research in the Life Sciences Hahn- Meitner-Istitut, Glienicker Str.100, 14109 Berlin A new world of tiny RNAs - siRNAs and miRNAs Exogenous phospholipases A2 mediate gene expression in rat mesangial cells via PPARalpha activation Cellular transformation mediated by Src kinase: the role of PTPalpha Investigations on the binding of SYBR Green I to double-stranded (ds)DNA vitamin E FUNCTIONAL SIGNIFICANCE OF GENETIC POLYMORPHISMS OF HUMAN CYP2B6 Quality control of membrane proteins in mitochondria The intracellular N-terminus of aminopeptidase N/CD13 affects association with caveolae Aminopeptidase N/CD13 is associated with Lubrol rafts Significance of cholesterol-independent effects of statins Elevated expression level of survivin protein in soft-tissue sarcomas Cytochrome P450-dependent eicosapentaenoic acid epoxygenation produces novel vasoactive metabolites Natural ligand identification and functional characterization of orphan G protein-coupled receptors SIGNALING AND TRANSCRIPTIONAL CONTROL OF VISCERAL MUSCLE AND HEART DEVELOPMENT IN DROSOPHILA P450non System from Acinetobacter sp. EB104: A Unique Bacterial Alkane Hydroxylase

Identification of the Major Tyrosin Phosphorylation Sites of Human Gab-1 Beta-adrenoceptor polymorphisms: does altered in vitro Leineweber, Kirsten function predict in vivo effects? The cardiac transcription factor Nkx2-5 regulates the alternative promoters of the endothelin-converting lemmer, julia enzyme-1 gene in H9c2 cardiomyoblasts Stability and Folding of Internalin B - A Biophysical Lengefeld, Jan Characterization p42/p44 MAP kinase (Erk) - Control of spatio-temporal LENORMAND, Philippe activity Effect of PFK-2 overexpression in insulin-producing cells Lenzen, Sigurd upon glucokinase activity and glucose metabolism Protein therapy: Applications in tissue regeneration and Leonhardt, Heinrich stem cell research OPTIMIZED ANTISENSE OLIGONUCLEOTIDE-MEDIATED INHIBITION OF THE INDUCIBLE NITRIC OXIDE Lerzynski, Guido SYNTHASE CO-SUPPRESSES NO-MEDIATED STRESS RESPONSE IN ENDOTHELIUM Functional organization of the yeast proteome by Leutwein, Christina systemic analysis of protein complexes ATP-dependent protein degradation and unfolding by Levchenko, I. ClpXP Identification of aminocoumarin antibiotics from different Li, Shuming Streptomyces species via ESI-MS and LC-MS/MS coupling CyGene: Eine neue hefebasierte Methode zur Bestimmung des cyto- und genotoxischen Potentials von Lichtenberg-Fraté, Hella Umweltgiften im Wasserbereich. CyGene: A New Yeast Based Method for the Detection of Cyto- and Genotox Cleavage of EWS/FLI-1 fusion transcripts by Liebig, Beate Hammerhead Ribozymes The Use of Multidimensional Capillary LC in Proteomics Liedtke, Steffen A mathematical model to predict plasma concentrations of morphine and its active metabolite Liefhold, Jürgen morphine-6-glucuronide in healthy young persons Functions of the proteasome in cell cycle and apoptosis Ligr, Martin GAF-domains as novel cAMP-sensor autoactivate a Linder, Jürgen bacterial adenylyl cyclase Signal transduction by SorLA, a member of the Lintzel, Julia LDL-receptor family Secretion of intact oligonucleotides by mrp2 into bile Lischka, Kerstin SIGNALING AND TRANSCRIPTIONAL CONTROL OF VISCERAL MUSCLE AND HEART DEVELOPMENT IN Lo, Patrick DROSOPHILA Optimal enzymes for single molecule sequencing Löbermann, Sylvia Promotion of hepatocarcinogenesis is mediated by overexpression of glutamine synthetase in Loeppen, Sandra beta-catenine-mutated mouse liver tumors Aldosterone action in humans -new members of the Loesel, Ralf aldosterone signaling pathwayBiomolecular switch from oxidative stress in inflammation to tissue morphogenesis in healing: Fenton-type OH* Logemann, Enno radical reactions and isoguanosine in metalloregulated RNA bioaptamers of S100-proteins Caspase inhibition strongly improves the recovery of los, marek cryopreserved hematopoietic and other cells Comparison of the effects of the P2Y12 ADP-receptor antagonist (AR-C69931) and the GPIIb/IIIa antagonist Lösche, Wolfgang GR 144053F on platelet-leukocyte conjugate formation. Differential effects of "advanced glycation endproducts" and ß-amyloid peptide on glucose utilization and ATP Loske, Claudia levels in the neuronal cell line SH-SY5Y A mathematical model to predict plasma concentrations of morphine and its active metabolite Lötsch, Jörn morphine-6-glucuronide in healthy young persons Characterization, Optimization and Application of Lu, XiaoLi Peptide-Mediated Protein Transduction Study on the function and kinetics of Lücke, Christian diisopropylfluorophosphatase Lehr, Stefan

Ludewig*, Maria

Sequence Analysis of an Acinetobacter Plasmid Carrying the Genes of a P450-Dependent Alkane Monooxygenase

The ratio of the cofactor NAD +/NADH of S-adenosylhomocysteine hydrolase controls the Angelika adenosine binding sites. CyGene: Eine neue hefebasierte Methode zur Bestimmung des cyto- und genotoxischen Potentials von Ludwig, Jost Umweltgiften im Wasserbereich. CyGene: A New Yeast Based Method for the Detection of Cyto- and Genotox Cytochrome P450-dependent eicosapentaenoic acid Luft, Friedrich C. epoxygenation produces novel vasoactive metabolites Cytochrome P450-Dependent Renal Metabolism of Arachidonic Acid in a Rat Model of Angiotensin II induced Luft, Friedrich C. Hypertension and End-Organ Damage Protein insertion into the inner membrane of Lutz, Tom mitochondria Optimization of intein based isotope labeling methods for Macek, Robert NMR investigations of proteins Stability and Folding of Internalin B - A Biophysical Machner, Matthias Characterization Appropriate TNF production - a decisive factor in sepsis Maennel, Daniela Differential regulation of gap junction protein expression Mahjour, Polin in cardiomyocytes Characterization, Optimization and Application of Mai, Jeff Peptide-Mediated Protein Transduction Redox events in IL-1 signaling Maiorino, Matilde SH2-domain containing inositol 5-phosphatase (SHIP)-1 is a potential regulator of cytoskeletal rearrangements in Mancini, Annalisa NGF induced neurite outgrowth Crystal structure of the noncollagenous (NC1) domain of human placenta collagen IV: Stabilization via a novel Mann, Karlheinz type of covalent Met-Lys cross-link Identification of an intermediate in the unfolding of neutral protease from Bacillus stearothermophilus by Mansfeld, Johanna fluorescence spectroscopy Generation of catalytically active neutral protease from Bacillus stearothermophilus does not require the presence Mansfeld, Johanna of the propeptide Differential subcellular distribution, structure and functions of the 14-3-3 proteins of the unicellular green Marquardt, Otfried alga Chlamydomonas reinhardtii Targeting of chimaeric GFP-polypeptide fusions within Marques, Joao Pedro chloroplasts Marques, Joao Pedro Import and plastid sorting of chimaeric GFP-polypeptides A new world of tiny RNAs - siRNAs and miRNAs Martinez, Javier Effect of PFK-2 overexpression in insulin-producing cells Massa, Maria Laura upon glucokinase activity and glucose metabolism Alkylphosphocholine Analogues induce Apoptosis in Massing, Ulrich Tumour Cells of the Immune System Alkylphosphocholine Analogues induce Apoptosis in Matzke, Astrid Tumour Cells of the Immune System FUNCTIONAL EXPRESSION OF HETEROLOGOUS Mauersberger, CYTOCHROME P450 IN YARROWIA LIPOLYTICA AND ITS Stephan USE IN BIOTRANSFORMATION OF STEROIDS Optimization of intein based isotope labeling methods for Maurer, Till NMR investigations of proteins Structure-function analysis of HscC, the E. coli member of Mayer, Matthias a novel subfamily of specialized Hsp70 chaperones Microarray analysis fo genes expressed in the frontal Mayer, Peter cortex of rats chronically treated with morphine The human inositol phosphate multikinase (IPMK) is targeted to the nucleus of mammalian cells by a basic Mayr, Georg cluster in its C-terminus cell cycle and death control: long live Forkheads Medema, Rene Protein insertion into the inner membrane of Meier, Stephan mitochondria Pharmacogenetic studies on drug-metabolizing cytochromes P450 and possibilities for individual drug Meisel, Christian therapy Mitochondrial translocases for precursor proteins Meisinger, Chris Lüdtke1,

Meister, Walter-Vesely Seb. Meixner, Silvia Meixner, Silvia Meltzer, P Menzel, Ralph Messerle, Martin Metzlaff, Michael Meye, Axel Meye, Axel Meyer, Helmut E. Meyer, Helmut E. Meyer, Helmut E. Meyer, Helmut E. Meyer, Jutta Meyer, Thomas F. Meyer*, Helmut E. Mi, Zhibao Michael, Schaupp Migeotte, Isabelle Mikhail, M. M. Molik, Sabine Monnerjahn, Christian Moriggl, Richard Moritz, Bernd Mühlberg, Katja Muller, Anja Müller, Claudia Müller, Cordula Müller, Dominik N. Müller, Günter Müller, Günter Müller, Helmut Müller, Meike Müller-Wieland, Dirk Münch, Gerald

Münch, Gerald

Polyvinylnucleobases: Synthesis, characterization and applications in nanotechnology Caspase-10 is the Key Initiator-Caspase involved in TBT-mediated Apoptosis Alkylphosphocholine Analogues induce Apoptosis in Tumour Cells of the Immune System Classification and diagnostic prediction of pediatric cancers using gene expression profiling and artificial neural networks Xenobiotically induced cytochrome P450 gene expression in Caenorhabditis elegans CYTOMEGALOVIRUS-A NEW VECTOR FOR GENE TRANSFER Transgene- and virus-mediated gene silencing - What plants can teach us! Elevated expression level of survivin protein in soft-tissue sarcomas Growth reduction of a xenotransplanted human Soft Tissue Sarcoma by MDM2 Antisense Therapy Quantitative Proteomics with Escherichia coli Analysis of Posttranslational Modifications of alpha-A-Crystallin during Aging of the Eye Lens Identification of the Major Tyrosin Phosphorylation Sites of Human Gab-1 Two-dimensional chromatography in protein analysis A new world of tiny RNAs - siRNAs and miRNAs Microarray analysis of mucosal gene expression profiles during gastric H. pylori infection High throughput mass spectrometry for proteomics Characterization, Optimization and Application of Peptide-Mediated Protein Transduction On the use of a histone-like protein for drug delivery using doxorubicin as a model system for tumor therapy Natural ligand identification and functional characterization of orphan G protein-coupled receptors Effect of Lacidipine on induced liver fibrosis / cirrhosis. Transport and assembly of the Rieske Fe/S protein: What happens in the stromal space? A designed variant of Herpes virus thymidine kinase selectively phosphorylates the anticancer and antiviral prodrug ganciclovir Activities of oncogenic STAT3 in colorectal cancer cells Quantitative Proteomics with Escherichia coli Differential regulation of gap junction protein expression in cardiomyocytes A role for Chemokine Receptors in Cancer Metastasis? The role of NCAM phosphorylation on NCAM mediated signal transduction pathways Redox events in IL-1 signaling Cytochrome P450-Dependent Renal Metabolism of Arachidonic Acid in a Rat Model of Angiotensin II induced Hypertension and End-Organ Damage A Receptor for Glycolipid Headgroups Mediates Insulin-mimetic Signaling in Rat Adipocytes Use of PTP1B mutants for selective inhibitor screening Hsp90 effects on the regulation of soluble guanylyl cyclase K-Ras-dependent signal tranduction in human pancreatic carcinoma cells Identification of the Major Tyrosin Phosphorylation Sites of Human Gab-1 Differential effects of "advanced glycation endproducts" and ß-amyloid peptide on glucose utilization and ATP levels in the neuronal cell line SH-SY5Y "Carbonyl stress” by methylglyoxal causes mitochondrial dysfunction, ATP depletion and NMDA receptor overstimulation in neurons

Münch, Gerald

Münch, Gerald Munck, Antonia Murad, Ferid Mürdter, Thomas E. Mürdter, Thomas E. Nagai, Yasuo Nagaso, Hideyuki Naim, H. Nalaskowski, Marcus Naumann, Anke Naumann, Michael Navarrete Santos, Alexander Navarrete Santos, Alexander Navarrete Santos, Alexander Navarrete Santos, Anne Nedvetsky, Pavel I. Neher, S. Neumann, Franz-Josef Ngezahajo, A. Nguyen-Xuan, Thai-Hoa Nieckchen, Petra Niederberger, Ellen Niederberger, Ellen

Niering, Petra Niering, Petra Nies, Anne T. Nißler, Ludwig Nöller, Joachim Nürnberger, Thorsten Oberle, Carolin

ß-amyloid peptide potentiates inflammatory responses induced by lipopolysaccharide, interferon-gamma and "advanced glycation endproducts" in a mouse microglia cell line Proinflammatory signal transduction pathways in N11 mouse microglial cell line activated by "Advanced Glycation Endproducts" Interactionpartners of SorLA Hsp90 effects on the regulation of soluble guanylyl cyclase FUNCTIONAL SIGNIFICANCE OF GENETIC POLYMORPHISMS OF HUMAN CYP2B6 ThioTEPA and Clopidogrel are specific mechanism-based inhibitors of human CYP2B6 Characterizing the signaling pathway of rat p18 amyloid beta (Aß) responsive protein (p18AßrP). SIGNALING AND TRANSCRIPTIONAL CONTROL OF VISCERAL MUSCLE AND HEART DEVELOPMENT IN DROSOPHILA Trafficking pathways of connexin49-GFP in living mammalian cells The human inositol phosphate multikinase (IPMK) is targeted to the nucleus of mammalian cells by a basic cluster in its C-terminus Effect of BMP-2 on the migratory and invasive properties of breast cancer cells Microarray analysis of mucosal gene expression profiles during gastric H. pylori infection The intracellular N-terminus of aminopeptidase N/CD13 affects association with caveolae Aminopeptidase N/CD13 is associated with Lubrol rafts Binding of RNA to the hyperthermophilic histone-like protein TmHU The insulin receptor and the insulin-dependent glucose transporter isoform 4 are expressed in rabbit preimplantation embryos Hsp90 effects on the regulation of soluble guanylyl cyclase ATP-dependent protein degradation and unfolding by ClpXP Molecular mechanisms of restenosis Trafficking pathways of connexin49-GFP in living mammalian cells The endosomal/lysosomal distribution of cathepsins (cat) B, L and S and N-acetyl-ß-D-glucosaminidase (NAG) is differentially modulated in murine cell lines of microglial and monocytic origin Optimal enzymes for single molecule sequencing Modulation of spinal nociceptive processing through the glutamate transporter GLT-1 Activation of c-Jun-N-terminal-kinase by R- and S-flurbiprofen results in cell cycle arrest in human colon carcinoma cells PRO- AND ANTIAPOPTOTIC EFFECTS OF QUERCETIN AND FISETIN IN H4IIE-CELLS: IMPLICATION OF OXIDATIVE STRESS H2O2-INDUCED OXIDATIVE STRESS IN H4IIE CELLS: INFLUENCE OF THE FLAVONOID KAEMPFEROL Proteins mediating vectorial transport across plasma membrane domains Binding of flavonoid and anthraquinone derivatives to nucleotide binding domains of ABC transporters. Membrane affinity and pore forming properties of proteins/peptides - Easy and fast by using solid supported lipid bilayers. Signal perception and transduction in the plant immune response Alkylphosphocholine Analogues induce Apoptosis in Tumour Cells of the Immune System

Odyvanova, Larissa Oehme, Gabriele Oesch, Franz Offermanns, Stefan Ohlenschläger, Oliver Ohly, Dorothea

Olbrich, Antje Ort, Stephan orzechowski, hans-dieter Orzechowski, Hans-Dieter Oselin, Kersti

Osswald1, Hartmut Ostareck, Dirk H. Otto, Henning Owen, David Pääbo+, Svante Paarmann, Ingo PAGES, Gilles Pamukci, Zuebeyde Parmentier, Marc Parthier, Christoph Patkaniowska, Agnieszka Patzelt, Holger paul, martin Paul, Martin Pauls, Regina

Paulsen, Reinhard Pesch, Beate Pesheva, Penka Petermann, Eva

Effect of BMP-2 on the migratory and invasive properties of breast cancer cells "Carbonyl stress” by methylglyoxal causes mitochondrial dysfunction, ATP depletion and NMDA receptor overstimulation in neurons Toxicology from experimental systems to man: drug metabolising enzymes as key determinants G-Protein mediated signalling: Studies in conditional Galpha knockouts Structure of an RNA Involved in Enteroviral Replication INDUCTION OF THE HUMAN NEURONAL NITRIC OXIDE SYNTHASE EXPRESSION BY EPIDERMAL GROWTH FACTOR IS MEDIATED THROUGH THE JAK/STAT SIGNAL TRANSDUCTION PATHWAY Effects of vitamin E on endothelial function in diabetes mellitus and hyperglycemia Structural and functional analysis of deoxyribonucleoside kinases dCK and dGK: design of novel mutant enzymes for therapeutic applications. The cardiac transcription factor Nkx2-5 regulates the alternative promoters of the endothelin-converting enzyme-1 gene in H9c2 cardiomyoblasts Effect of MAP kinase inhibitors on injury-induced neointima formation in the rat carotid artery Pharmacogenetic studies on drug-metabolizing cytochromes P450 and possibilities for individual drug therapy The ratio of the cofactor NAD +/NADH of S-adenosylhomocysteine hydrolase controls the adenosine binding sites. Translational activation from DICE-mediated mRNA silencing by the Src tyrosine kinase Characterization of Inner Nuclear Membrane Proteins Localisation of fluorescently labelled Ras protein in the living cell Sequence Analysis of an Acinetobacter Plasmid Carrying the Genes of a P450-Dependent Alkane Monooxygenase Structural requirements for calmodulin binding to membrane-associated guaylate kinases p42/p44 MAP kinase (Erk) - Control of spatio-temporal activity Listeriolysin of Listeria monocytogenes forms Ca2+-permeable pores leading to intracellular Ca2+ oscillations in host cells Natural ligand identification and functional characterization of orphan G protein-coupled receptors Polyomavirus-like particles as a system for gene therapy and drug delivery A new world of tiny RNAs - siRNAs and miRNAs Structure-function analysis of HscC, the E. coli member of a novel subfamily of specialized Hsp70 chaperones The cardiac transcription factor Nkx2-5 regulates the alternative promoters of the endothelin-converting enzyme-1 gene in H9c2 cardiomyoblasts Effect of MAP kinase inhibitors on injury-induced neointima formation in the rat carotid artery Efficient Dosage and Time dependent Protein Transduction of Pancreatic Carcinoma Cells using purified VP22-EGFP Fusion Protein The INAD signaling complex of Drosophila photoreceptor: Recombinant Expression and Assembly in a Cell Culture System Breast Cancer and Predictive Factors: Association with Polymorphisms Tenascins are associated with lipid rafts isolated from mouse brain Generation of catalytically active neutral protease from Bacillus stearothermophilus does not require the presence of the propeptide

Peterson, C Petry, Stefan Petry, Stefan Petzinger, Ernst Pfanner, Nikolaus Pfanner, Nikolaus Pfeifer, Dietmar Pfeiffer, Dietrich Pfeil, Andrea Pfeil, Wolfgang Pfeilschifter, Josef Pfitzner, Edith Pfluger, Paul PHAN, Gia Anh Bao

PHAN, Gia Anh Bao

PIETROWSKI, Detlef

PIETROWSKI, Detlef

Pineda, Miguel Plückthun, Andreas Pocar, Paola Pojer, Florence Polesskaya, Anna Poppe, Monika Porzig, Hartmut POUYSSEGUR, Jacques Preuss, Marc Prisner, Thomas Prösch, Susanna

Racké, Kurt Radons, Jürgen Ramachandran, Ramadurai Rapoport, Tom

Classification and diagnostic prediction of pediatric cancers using gene expression profiling and artificial neural networks A Receptor for Glycolipid Headgroups Mediates Insulin-mimetic Signaling in Rat Adipocytes Use of PTP1B mutants for selective inhibitor screening Secretion of intact oligonucleotides by mrp2 into bile Mitochondrial translocases for precursor proteins The TIM22 translocase mediates insertion of carrier precursors into the mitochondrial inner membrane via a two-step process Pharmacogenetic profiling with Biochips: an innovative concept to improve the efficiency of clinical trials Differential regulation of gap junction protein expression in cardiomyocytes Evolution of Feedback-inhibited (&beta/&alpha)8 Barrel Isoenzymes by Gene Duplication and A Single Mutation Stability and Folding of Internalin B - A Biophysical Characterization Exogenous phospholipases A2 mediate gene expression in rat mesangial cells via PPARalpha activation Activities of oncogenic STAT3 in colorectal cancer cells vitamin E THE EXPRESSION OF ANGIOGENIC GROWTH FACTORS IN HUMAN GRANULOSA CELLS ARE CONTROLLED BY HUMAN CHORION GONADOTROPIN. EXPRESSION OF A NEW SPLICE VARIANT OF IGFBP-7/MAC25 TUMOR SUPPRESSOR GENE IN HUMAN GRANULOSA CELLS LACKING THE IGFBP MOTIF (GCGCCXXC) THE EXPRESSION OF ANGIOGENIC GROWTH FACTORS IN HUMAN GRANULOSA CELLS ARE CONTROLLED BY HUMAN CHORION GONADOTROPIN. EXPRESSION OF A NEW SPLICE VARIANT OF IGFBP-7/MAC25 TUMOR SUPPRESSOR GENE IN HUMAN GRANULOSA CELLS LACKING THE IGFBP MOTIF (GCGCCXXC) GAF-domains as novel cAMP-sensor autoactivate a bacterial adenylyl cyclase Novel proteins from combinatorial and evolutionary approaches Induction of apoptotic cell death in bovine cumulus-oocyte complexes after treatment with Aroclor-1254 during in vitro maturation. Identification of aminocoumarin antibiotics from different Streptomyces species via ESI-MS and LC-MS/MS coupling Myogenic Specification of Adult Stem Cells in Skeletal Muscle Bcl-2 antagonist of cell death (Bad) retains its proapoptotic activity despite proteolytic cleavage by caspase-3 SDF-1 and thrombin signaling in human hematopoietic progenitor cells is regulated by cytokines p42/p44 MAP kinase (Erk) - Control of spatio-temporal activity Protein insertion into the inner membrane of mitochondria Are GTP-analogs really good analogs? From the molecular mechanism of human cytomegalovirus (re)activation to new therapeutical concepts Susanna Prösch and Hans-Dieter Volk Institutes of Virology and Medical Immunology, Humboldt Unive Reduced NO synthesis during halothane exposure in stimulated rat alveolar macrophages Molecular modeling as a tool to identify putative interacting sites in the IL-1 receptor complex Structure of an RNA Involved in Enteroviral Replication Protein Transport In and Out of the ER

Rauch, Thomas Rauch, Uwe Real, Robert Reents, Reinhard Rehling, Peter Rehling, Peter Rehling, Peter Reichert, Kerstin Reim, Ingolf Reinders, Jörg Reis, Andrè Reiser, Georg Reiser, Georg

Repp, Holger Reuss, Matthias Ricarda, Thier Richardt*, André Richter, Günther Richter, Günther Richter, Jan Richter, Tanja Richter, Tanja Riemann, Dagmar Riemann, Dagmar Ries, Albert

Ringner, M. Rist, Wolfgang Rita, Bernhardt Robbins, Paul Robinson, Colin Rochon, Maike Rödel, Matthias Roden, Dan

Cytosolic proteins at birth: linking translation and chaperone assisted protein folding Tenascins are associated with lipid rafts isolated from mouse brain Effect of MAP kinase inhibitors on injury-induced neointima formation in the rat carotid artery Localisation of fluorescently labelled Ras protein in the living cell Mitochondrial translocases for precursor proteins The TIM22 translocase mediates insertion of carrier precursors into the mitochondrial inner membrane via a two-step process Molecular Dissection of the Protein Import Complexes of the Inner Membrane of Yeast Mitochondria Xenobiotically induced cytochrome P450 gene expression in Caenorhabditis elegans SIGNALING AND TRANSCRIPTIONAL CONTROL OF VISCERAL MUSCLE AND HEART DEVELOPMENT IN DROSOPHILA Identification of the Major Tyrosin Phosphorylation Sites of Human Gab-1 Understanding the genetic basis of common human diseases and its potential for new concepts in drug therapy Regulation of docosahexaenoic and arachidonic acid release in rat astrocytes Different regulation of human lung cancer cell proliferation by nucleotides mediated through P2Y receptors Listeriolysin of Listeria monocytogenes forms Ca2+-permeable pores leading to intracellular Ca2+ oscillations in host cells Quantitative Proteomics with Escherichia coli Enzym polymorphisms and risk factors in bladder cancer patients in Lutherstadt Wittenberg Study on the function and kinetics of diisopropylfluorophosphatase Nasal DNA vaccination with an Interleukin-10 cDNA vector in a murine asthma modell Differential analysis of interleukin 2 (IL-2) versus IL-15 induced immediate-early genes: A contribution for the elucidation of the AICD paradoxon? Nitrogen oxide-induced cell death in rabbit cultured Müller cells is modulated by glucose FUNCTIONAL SIGNIFICANCE OF GENETIC POLYMORPHISMS OF HUMAN CYP2B6 ThioTEPA and Clopidogrel are specific mechanism-based inhibitors of human CYP2B6 The intracellular N-terminus of aminopeptidase N/CD13 affects association with caveolae Aminopeptidase N/CD13 is associated with Lubrol rafts Crystal structure of the noncollagenous (NC1) domain of human placenta collagen IV: Stabilization via a novel type of covalent Met-Lys cross-link Classification and diagnostic prediction of pediatric cancers using gene expression profiling and artificial neural networks Cytosolic proteins at birth: linking translation and chaperone assisted protein folding Activities of CYP11B isoforms from different species. Characterization, Optimization and Application of Peptide-Mediated Protein Transduction Structure and mechanism of the twin-arginine translocase Targeting of proteins to the flagellar type III secretion system of Escherichia coli Xenobiotically induced cytochrome P450 gene expression in Caenorhabditis elegans Implementing pharmacogenomics to improve drug therapy: where do we stand in 2002

Pharmacogenetic studies on drug-metabolizing cytochromes P450 and possibilities for individual drug therapy Effects of vitamin E on endothelial function in diabetes Rösen, Peter mellitus and hyperglycemia Chaperons at the Ribosomal Tunnel Exit - First Contacts Rospert, Sabine of a Newly Synthesized Polypeptide Modulation of spinal nociceptive processing through the Rothstein, Jeffrey D. glutamate transporter GLT-1 The peptide-binding domain determines the interaction of mitochondrial Hsp70s with the essential partner protein Röttgers, Karin Tim44 Multivalent Scavengers of Oncogenic Ras: A novel approach to tracking and therapeutical inhibition of Ras Rubio, Ignacio activity Myogenic Specification of Adult Stem Cells in Skeletal Rudnicki, Michael Muscle vitamin E Rühl, Ralph Study on the function and kinetics of Rüterjans, Heinz diisopropylfluorophosphatase Classification and diagnostic prediction of pediatric cancers using gene expression profiling and artificial Saal, L. H. neural networks Involvement of La protein in alternative translation Saaler-Reinhardt, Sigrid processes in neural progenitor cells Catechins with a galloyl group in the 3-position inhibit the Sachinidis, Agapios PDGF-BB-induced intracellular signal transduction pathway and proliferation of vascular smooth muscle cells PDGF-BB induces selective differentiation of mouse Sachinidis, Agapios embryonic stem cells to cardiac cells Differential regulation of gap junction protein expression Salameh, Aida in cardiomyocytes Caspase inhibition strongly improves the recovery of samraj, ajoy cryopreserved hematopoietic and other cells Use of cytochrome P450 isoenzyme assays in the Sander, Uta hit-to-lead process of drug discovery Putative role of the nuclear factor NCNF in neuronal Sattler, Ulrike differentiation ATP-dependent protein degradation and unfolding by Sauer, R.T. ClpXP Characterizing the signaling pathway of rat p18 amyloid Sawada, Tohru beta (Aß) responsive protein (p18AßrP). Analysis of Posttranslational Modifications of Schaefer, Heike alpha-A-Crystallin during Aging of the Eye Lens High throughput mass spectrometry for proteomics Schaefer, Heike Different regulation of human lung cancer cell proliferation by nucleotides mediated through P2Y Schäfer, Rainer receptors Phospholipase D from cabbage: Calcium ions essential for Schäffner, Ines activity destabilize the protein conformation INHIBITION OF EROD-ACTIVITY IN RAT HEPATOMA Scharmm, Karl-Werner CELLS Targeting of chimaeric GFP-polypeptide fusions within Schattat, Martin chloroplasts Import and plastid sorting of chimaeric GFP-polypeptides Schattat, Martin Signal perception and transduction in the plant immune Scheel, Dierk response Nasal DNA vaccination with an Interleukin-10 cDNA Scherf, Wiebke vector in a murine asthma modell Activation of c-Jun-N-terminal-kinase by R- and S-flurbiprofen results in cell cycle arrest in human colon Schilling, Karin carcinoma cells Tenascins are associated with lipid rafts isolated from Schilling, Karl mouse brain Fanconi anemia: basic molecular biology and clinical Schindler, Detlev implications Advanced Glycation Endproducts induced cell signalling in Schinzel, Reinhard cardiac fibroblasts Interaction of the organic base cimetidine with the renal Schlattjan, Jan Henrik basolateral p-aminohippurate (PAH) transport system Roots, Ivar

Schlichting, Ilme Schlomann, Uwe Schlott, Bernhard Schlüter, Thomas

Schmid, Heide

Schmidt, Hannelore Schmidt, Hannelore Schmidt, Harald H.H.W. Schmidt, Helmut Schmidt, Ulrich Schmidtko, Achim Schmitt, Johannes

Schmitt, Marcel

Schmitz, Brigitte Schmitz, Brigitte Schmitz, Brigitte Schmitz, Brigitte Schmitz, Brigitte

Schneider, Christina¹

Schneider, Daniela Schneider, Thomas Schnurr, Kerstin Schön, Astrid Schönfelder, Manfred Schramm, Hans J. Schreckenberger, Susan Schröder, Peter Schröder-Borm, Hannah Schultheis, Martina Schultz, Anita Schultz, Joachim Schulz, Burkhard

Crystallographic studies on the reaction mechanism of P450 Tumor necrosis factor alpha signalling in reactive glia cells p53 is involved in the repair of human topoisomerase I cleavage complexes Characterisation of Protein Domains Responsible for Recognition of P-Selectin by Sorting Nexin 17 The endosomal/lysosomal distribution of cathepsins (cat) B, L and S and N-acetyl-ß-D-glucosaminidase (NAG) is differentially modulated in murine cell lines of microglial and monocytic origin Elevated expression level of survivin protein in soft-tissue sarcomas Growth reduction of a xenotransplanted human Soft Tissue Sarcoma by MDM2 Antisense Therapy Hsp90 effects on the regulation of soluble guanylyl cyclase A mathematical model to predict plasma concentrations of morphine and its active metabolite morphine-6-glucuronide in healthy young persons Polyomavirus-like particles as a system for gene therapy and drug delivery Modulation of spinal nociceptive processing through the glutamate transporter GLT-1 Membrane affinity and pore forming properties of proteins/peptides - Easy and fast by using solid supported lipid bilayers. CyGene: Eine neue hefebasierte Methode zur Bestimmung des cyto- und genotoxischen Potentials von Umweltgiften im Wasserbereich. CyGene: A New Yeast Based Method for the Detection of Cyto- and Genotox The role of NCAM phosphorylation on NCAM mediated signal transduction pathways The role of serine-phosphorylation of the cell adhesion molecule L1 in basal neurite outgrowth The O-GlcNAc modification of proteins interferes with signaling by PKA and CDK5 dependent phosphorylation in neurons Function of the PEST sequence of NCAM 140 Altered O-GlcNAc level after neuronal differentiation of PC12 cells Inverse Dynamics of Glial Fibrillary Acidic Protein (GFAP) Expression and Synthesis of Erythropoietin (EPO), EPO-Receptor (EPOR) and Proliferating Cell Nuclear Antigen (PCNA) in Astrocytes Promotion of hepatocarcinogenesis is mediated by overexpression of glutamine synthetase in beta-catenine-mutated mouse liver tumors Evolution of Feedback-inhibited (&beta/&alpha)8 Barrel Isoenzymes by Gene Duplication and A Single Mutation Redox events in IL-1 signaling The evolution of RNase P in eukaryotic organelles Growth reduction of a xenotransplanted human Soft Tissue Sarcoma by MDM2 Antisense Therapy Dimerization inhibitors of HIV-1 protease Characterisation of Protein Domains Responsible for Recognition of P-Selectin by Sorting Nexin 17 (-)-Epicatechin Is Taken up by Cells and Protects against Peroxynitrite-Induced Oxidation and Nitration. Molecular basis for membrane selectivity of a potent peptide antibiotic derived from NK-lysin The role of serine-phosphorylation of the cell adhesion molecule L1 in basal neurite outgrowth GAF-domains as novel cAMP-sensor autoactivate a bacterial adenylyl cyclase GAF-domains as novel cAMP-sensor autoactivate a bacterial adenylyl cyclase Isolation of an Arabidopsis T-DNA mutant for the chloroplast protein import receptor atToc33

GLUTATHIONE S-TRANSFERASE GSTP1 POLYMORPHISMS IN DE NOVO AND THERAPY-INDUCED HEMATOLOGIC MALIGNANCIES Bcl-2 antagonist of cell death (Bad) retains its Schulze-Osthoff, Klaus proapoptotic activity despite proteolytic cleavage by caspase-3 Caspase inhibition strongly improves the recovery of schulze-osthoff, klaus cryopreserved hematopoietic and other cells Cytochrome P450-Dependent Renal Metabolism of Arachidonic Acid in a Rat Model of Angiotensin II induced Schunck, Wolf-H. Hypertension and End-Organ Damage Cytochrome P450-dependent eicosapentaenoic acid Schunck, Wolf-Hagen epoxygenation produces novel vasoactive metabolites Metabolism of eicosapentaenoic acid by CYP4A and CYP2C Schunck, Wolf-Hagen enzymes A designed variant of Herpes virus thymidine kinase selectively phosphorylates the anticancer and antiviral Schüttmann, Ina prodrug ganciclovir Classification and diagnostic prediction of pediatric cancers using gene expression profiling and artificial Schwab, M. neural networks FUNCTIONAL SIGNIFICANCE OF GENETIC Schwab, Matthias POLYMORPHISMS OF HUMAN CYP2B6 ThioTEPA and Clopidogrel are specific mechanism-based Schwab, Matthias inhibitors of human CYP2B6 Effect of BMP-2 on the migratory and invasive properties Schwalbe, Manuela of breast cancer cells The endosomal/lysosomal distribution of cathepsins (cat) B, L and S and N-acetyl-ß-D-glucosaminidase (NAG) is Schwarz, Gerold differentially modulated in murine cell lines of microglial and monocytic origin Bcl-2 antagonist of cell death (Bad) retains its proapoptotic activity despite proteolytic cleavage by Schwarz, Michael caspase-3 Promotion of hepatocarcinogenesis is mediated by overexpression of glutamine synthetase in Schwarz, Michael beta-catenine-mutated mouse liver tumors Sequence Analysis of an Acinetobacter Plasmid Carrying Schwarz+, Carsten the Genes of a P450-Dependent Alkane Monooxygenase Use of PTP1B mutants for selective inhibitor screening Schwenk, Robert Myogenic Specification of Adult Stem Cells in Skeletal Scime, Anthony Muscle Myogenic Specification of Adult Stem Cells in Skeletal Seale, Patrick Muscle Stability and Folding of Internalin B - A Biophysical Seckler, Robert Characterization Pectate Lyase from Bacillus subtilis (BsPel): Mutations in the asparagine ladder cause a temperature-sensitive Seckler, Robert folding phenotype Folding Kinetics of Pectate Lyase from Bacillus subtilis Seckler, Robert Refolding from Fragments: The case of the proteolytically Seckler, Robert processed Pea Lectin The E.coli membrane insertase YidC: A Chaperone for Serek, Justyna Protein Folding into the Membrane GRIF-1 - a novel GABAA receptor associated protein Sharma, Seema Analysis of Posttranslational Modifications of Sickmann, Albert alpha-A-Crystallin during Aging of the Eye Lens Identification of the Major Tyrosin Phosphorylation Sites Sickmann, Albert of Human Gab-1 High throughput mass spectrometry for proteomics Sickmann, Albert Two-dimensional chromatography in protein analysis Sickmann, Albert Pharmacogenetics in the Cardiovascular Field Siegmund, Werner Quantitative Proteomics with Escherichia coli Siemann, Martin (-)-Epicatechin Is Taken up by Cells and Protects against Sies, Helmut Peroxynitrite-Induced Oxidation and Nitration. Activation of EGF receptor-dependent signaling pathways Sies, Helmut by alkylating and redox-cycling quinones Advanced Glycation Endproducts induced cell signalling in Silber, Rolf-Edgar cardiac fibroblasts Activities of CYP11B isoforms from different species. Silvia, Zearo Schulz, Thomas

Simm, Andreas Simon, Hans-Uwe Simons, Kai Skach, Romanita

Skarke, Carsten Smith, Miriam

Advanced Glycation Endproducts induced cell signalling in cardiac fibroblasts MACROPHAGE MIGRATION INHIBITORY FACTOR (MIF) IS A SURVIVAL FACTOR FOR NEUTROPHILS Lipid rafts in membrane trafficking and cell polarity Catechins with a galloyl group in the 3-position inhibit the PDGF-BB-induced intracellular signal transduction pathway and proliferation of vascular smooth muscle cells A mathematical model to predict plasma concentrations of morphine and its active metabolite morphine-6-glucuronide in healthy young persons GRIF-1 - a novel GABAA receptor associated protein

p53 is involved in the repair of human topoisomerase I cleavage complexes Comparison of the effects of the P2Y12 ADP-receptor Spangenberg, Peter antagonist (AR-C69931) and the GPIIb/IIIa antagonist GR 144053F on platelet-leukocyte conjugate formation. Pharmacogenetic profiling with Biochips: an innovative Spleiss, Olivia concept to improve the efficiency of clinical trials Are GTP-analogs really good analogs? Spoerner, Michael Analysis of EWS/Fli-1 induced gene expression in HEK293 Staege, Martin S. cells using DNA-microarrays From genes to antigens: Potential target structures for autologous and allogeneic cytotoxic T cells identified by Staege, Martin S. oligonucleotide arrays Cleavage of EWS/FLI-1 fusion transcripts by Staege, Martin S. Hammerhead Ribozymes Secretion of intact oligonucleotides by mrp2 into bile Starke, Dieter Transcription of Reporter Genes with Immobilized Steffen, Jenny Templates MRP expression and modulation of/by glutathione in Stehfest, Ekkehardt human lung cells Are GTP-analogs really good analogs? Steiner, Guido Bcl-2 antagonist of cell death (Bad) retains its Stepczynska, Anna proapoptotic activity despite proteolytic cleavage by caspase-3 p53 is involved in the repair of human topoisomerase I Stephan, Holger cleavage complexes GRIF-1 - a novel GABAA receptor associated protein Stephenson, F. Anne Differential subcellular distribution, structure and functions of the 14-3-3 proteins of the unicellular green Stevanovic, Stefan alga Chlamydomonas reinhardtii Use of cytochrome P450 isoenzyme assays in the Stevens, Susanne hit-to-lead process of drug discovery p53 is involved in the repair of human topoisomerase I Stevnsner*, Tinna cleavage complexes Molecular modelling of HIV regulatory proteins as targets Sticht, Heinrich of viral infectivity Söe, Kent

Stoeva2, Stanka Stoll, Monika Strack, Siegfried

strasdat, meike Strasdat, Meike stroh, christopher Strokin, Mikhail Strub, Andreas Superti-Furga, Giulio

The ratio of the cofactor NAD +/NADH of S-adenosylhomocysteine hydrolase controls the adenosine binding sites. Genetic screening approaches for the identification of cardiovascular drug targets Involvement of different signalling pathways in cytoskeletal reorganisation during tributyltin (TBT) and H2O2 induced apoptosis The cardiac transcription factor Nkx2-5 regulates the alternative promoters of the endothelin-converting enzyme-1 gene in H9c2 cardiomyoblasts Effect of MAP kinase inhibitors on injury-induced neointima formation in the rat carotid artery Caspase inhibition strongly improves the recovery of cryopreserved hematopoietic and other cells Regulation of docosahexaenoic and arachidonic acid release in rat astrocytes The peptide-binding domain determines the interaction of mitochondrial Hsp70s with the essential partner protein Tim44 Functional organization of the yeast proteome by systemic analysis of protein complexes

OPTIMIZED ANTISENSE OLIGONUCLEOTIDE-MEDIATED INHIBITION OF THE INDUCIBLE NITRIC OXIDE Suschek, Christoph V. SYNTHASE CO-SUPPRESSES NO-MEDIATED STRESS RESPONSE IN ENDOTHELIUM Protein insertion into the inner membrane of Szyrach, Gregor mitochondria SH2-domain containing inositol 5-phosphatase (SHIP)-1 is a potential regulator of cytoskeletal rearrangements in Tamura, Teruko NGF induced neurite outgrowth The role of co- and posttranslational modifications in the Tatzelt, Jörg propagation of scrapie-prions Elevated expression level of survivin protein in soft-tissue Taubert, Helge sarcomas Growth reduction of a xenotransplanted human Soft Taubert, Helge Tissue Sarcoma by MDM2 Antisense Therapy Modulation of spinal nociceptive processing through the Tegeder, Irmgard glutamate transporter GLT-1 Activation of c-Jun-N-terminal-kinase by R- and S-flurbiprofen results in cell cycle arrest in human colon Tegeder, Irmgard carcinoma cells Function of the PEST sequence of NCAM 140 Tenderich, Reinhild A Receptor for Glycolipid Headgroups Mediates Tennagels, Norbert Insulin-mimetic Signaling in Rat Adipocytes Use of PTP1B mutants for selective inhibitor screening Tennagels, Norbert Crystal structure of the noncollagenous (NC1) domain of human placenta collagen IV: Stabilization via a novel Than, Manuel E. type of covalent Met-Lys cross-link Cytochrome P450-dependent eicosapentaenoic acid Theuer, Jürgen epoxygenation produces novel vasoactive metabolites Cytochrome P450-Dependent Renal Metabolism of Arachidonic Acid in a Rat Model of Angiotensin II induced Theuer, Jürgen Hypertension and End-Organ Damage Functional lipidomics Thiele, Christoph Enzym polymorphisms and risk factors in bladder cancer Thilo, Seidel patients in Lutherstadt Wittenberg The cardiac transcription factor Nkx2-5 regulates the alternative promoters of the endothelin-converting thomas, alexander enzyme-1 gene in H9c2 cardiomyoblasts Aut10p, Mai1p and Aut5p shed new lights on the Thumm, Michael molecular mechanisms of autophagy in S.cerevisiae Effect of PFK-2 overexpression in insulin-producing cells Tiedge, Markus upon glucokinase activity and glucose metabolism Crystal structure of the noncollagenous (NC1) domain of human placenta collagen IV: Stabilization via a novel Timpl, Rupert type of covalent Met-Lys cross-link The insulin receptor and the insulin-dependent glucose transporter isoform 4 are expressed in rabbit Tonack, Sarah preimplantation embryos MRP expression and modulation of/by glutathione in Torky, Abdel human lung cells The endosomal/lysosomal distribution of cathepsins (cat) B, L and S and N-acetyl-ß-D-glucosaminidase (NAG) is Tran, Minh-Huy differentially modulated in murine cell lines of microglial and monocytic origin PRO- AND ANTIAPOPTOTIC EFFECTS OF QUERCETIN AND Tran-Thi, Quynh-Hoa FISETIN IN H4IIE-CELLS: IMPLICATION OF OXIDATIVE STRESS H2O2-INDUCED OXIDATIVE STRESS IN H4IIE CELLS: Tran-Thi, Quynh-Hoa INFLUENCE OF THE FLAVONOID KAEMPFEROL Bcl-2 antagonist of cell death (Bad) retains its proapoptotic activity despite proteolytic cleavage by Troppmair, Jakob caspase-3 GLUTATHIONE S-TRANSFERASE GSTP1 POLYMORPHISMS IN DE NOVO AND THERAPY-INDUCED HEMATOLOGIC Trümper, Lorenz MALIGNANCIES Mitochondrial translocases for precursor proteins Truscott, Kaye The TIM22 translocase mediates insertion of carrier precursors into the mitochondrial inner membrane via a Truscott, Kaye, N. two-step process Activities of oncogenic STAT3 in colorectal cancer cells Tsareva, Svetlana

Tuschl, Thomas Ulbrich-Hofmann, Renate Ulbrich-Hofmann, Renate Ulbrich-Hofmann, Renate Vakili, Jalal van den Berg, Lambertus Veith, Anne-Marie Velten, Iris Vichalkovski, Anton Vitzthum, Frank Voegel, Thomas

Voigt, Jürgen Volff, Jean-Nicolas

Volk, Hans-Dieter

Völkel, Katja VOLMAT, Veronique von Figura, Kurt von Figura, Kurt von langsdorff, christian von Montfort, Claudia von Nickisch-Rosenegk, Markus von Pall de Tolna, Susanne

A new world of tiny RNAs - siRNAs and miRNAs Identification of an intermediate in the unfolding of neutral protease from Bacillus stearothermophilus by fluorescence spectroscopy Generation of catalytically active neutral protease from Bacillus stearothermophilus does not require the presence of the propeptide Phospholipase D from cabbage: Calcium ions essential for activity destabilize the protein conformation Natural ligand identification and functional characterization of orphan G protein-coupled receptors The E.coli membrane insertase YidC: A Chaperone for Protein Folding into the Membrane DMRT Genes and Sex Determination in the Fish Xiphophorous Functions of the proteasome in cell cycle and apoptosis SDF-1 and thrombin signaling in human hematopoietic progenitor cells is regulated by cytokines Investigations on the binding of SYBR Green I to double-stranded (ds)DNA Bcl-2 antagonist of cell death (Bad) retains its proapoptotic activity despite proteolytic cleavage by caspase-3 Differential subcellular distribution, structure and functions of the 14-3-3 proteins of the unicellular green alga Chlamydomonas reinhardtii DMRT Genes and Sex Determination in the Fish Xiphophorous From the molecular mechanism of human cytomegalovirus (re)activation to new therapeutical concepts Susanna Prösch and Hans-Dieter Volk Institutes of Virology and Medical Immunology, Humboldt Unive Oxidative stress in rat alveolar epithelial cells and co-cultures upon treatment with fly ash p42/p44 MAP kinase (Erk) - Control of spatio-temporal activity C(alpha)-Formylglycine: a novel protein modification in pro- and eukaryotes Inherited disorders caused by faulty protein modification in the secretory route The cardiac transcription factor Nkx2-5 regulates the alternative promoters of the endothelin-converting enzyme-1 gene in H9c2 cardiomyoblasts Singlet Molecular Oxygen Reversibly Inhibits Tyrosine Phosphatases Transcription of Reporter Genes with Immobilized Templates

Restriction endonuclease EcoRI - fusions for a change of specificity Restriction endonuclease EcoRI - fusions for a change of von Witzendorff, Dorothee specificity The peptide-binding domain determines the interaction of mitochondrial Hsp70s with the essential partner protein Voos, Wolfgang Tim44 Cytosolic proteins at birth: linking translation and Vorderwülbecke, Sonja chaperone assisted protein folding Localisation of fluorescently labelled Ras protein in the Wagner, Melanie living cell The TIM22 translocase mediates insertion of carrier precursors into the mitochondrial inner membrane via a Wagner, Richard two-step process High throughput mass spectrometry for proteomics Wagner, Yvonne Two-dimensional chromatography in protein analysis Wagner, Yvonne ATP-dependent protein degradation and unfolding by Wah, D. ClpXP Localisation of fluorescently labelled Ras protein in the Waldmann, Herbert living cell Microarray analysis of mucosal gene expression profiles Walduck, Anna during gastric H. pylori infection

Walter, Monika

Walter, Monika Walter, Monika Warnke**, J.P.

Wätjen, Wim Wätjen, Wim Wätjen, Wim Weber, Klaus Weber-Sparenberg, Corinna Wehling, Martin Wei, J. S. Weinfurtner, Daniel Weinstrauch, Anne Welte, Stefan Wenzel, Folker Werner, Sabine Westermann, F. Wettschureck, Nina Wied, Susanne Wiedemann, Nils Wiedenmann, Bertram Wiedenmann, Bertram Wiederanders, Bernd Wiegand, Christiane

Wiesinger, Heiner²

Wiesinger, Heinrich

Wilde, Christian Wilde, Christian Wildeboer, Dirk Wilkinson, Helen Willumeit, Regine Wilms, Regina

Pectate Lyase from Bacillus subtilis (BsPel): Mutations in the asparagine ladder cause a temperature-sensitive folding phenotype Pectate Lyase from Bacillus subtilis (BsPel): Mutations in the asparagine ladder cause a temperature-sensitive folding phenotype Folding Kinetics of Pectate Lyase from Bacillus subtilis Expression of ubiquitous 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase in intracranial neoplasms and non-tumorous brain tissue PRO- AND ANTIAPOPTOTIC EFFECTS OF QUERCETIN AND FISETIN IN H4IIE-CELLS: IMPLICATION OF OXIDATIVE STRESS H2O2-INDUCED OXIDATIVE STRESS IN H4IIE CELLS: INFLUENCE OF THE FLAVONOID KAEMPFEROL ALTERED SENSITIVITY OF TNF-ALPHA OVEREXPRESSING RAT HEPATOMA CELLS AGAINST EXOGENOUS TNF-ALPHA AND OXIDANTS A new world of tiny RNAs - siRNAs and miRNAs Targeting of proteins to the flagellar type III secretion system of Escherichia coli Aldosterone action in humans -new members of the aldosterone signaling pathwayClassification and diagnostic prediction of pediatric cancers using gene expression profiling and artificial neural networks Optimization of intein based isotope labeling methods for NMR investigations of proteins Effect of MAP kinase inhibitors on injury-induced neointima formation in the rat carotid artery Use of PTP1B mutants for selective inhibitor screening Reduced NO synthesis during halothane exposure in stimulated rat alveolar macrophages Altered O-GlcNAc level after neuronal differentiation of PC12 cells Classification and diagnostic prediction of pediatric cancers using gene expression profiling and artificial neural networks G-Protein mediated signalling: Studies in conditional Galpha knockouts A Receptor for Glycolipid Headgroups Mediates Insulin-mimetic Signaling in Rat Adipocytes Mitochondrial translocases for precursor proteins Microarray analysis of mucosal gene expression profiles during gastric H. pylori infection Educational aspects of molecular medicine in basic science and clinical training Activities of oncogenic STAT3 in colorectal cancer cells Temperature-sensitive TMV Coat Proteins as Pathogens in Animal Cells Inverse Dynamics of Glial Fibrillary Acidic Protein (GFAP) Expression and Synthesis of Erythropoietin (EPO), EPO-Receptor (EPOR) and Proliferating Cell Nuclear Antigen (PCNA) in Astrocytes The endosomal/lysosomal distribution of cathepsins (cat) B, L and S and N-acetyl-ß-D-glucosaminidase (NAG) is differentially modulated in murine cell lines of microglial and monocytic origin Clostridium botulinum C3 exoezyme like ADP-ribosyltransferases Clostridium botulinum C2 toxin as a protein transport system Tumor necrosis factor alpha signalling in reactive glia cells GRIF-1 - a novel GABAA receptor associated protein Molecular basis for membrane selectivity of a potent peptide antibiotic derived from NK-lysin SH2-domain containing inositol 5-phosphatase (SHIP)-1 is a potential regulator of cytoskeletal rearrangements in NGF induced neurite outgrowth

Winklhofer, Konstanze F. Wissler, Josef H.

Wissler, Josef H.

WITT, Paulina Wittamer, Valérie Wittinghofer, Alfred Wittinghofer, Alfred Wittko, Ina Wöhnert, Jens Wolf, Alexander Wölfl, Stefan Wulfaenger, Jens Wulfänger, Jens Wunder, Christian Würl, Peter Würl, Peter Yalcin, Abdullah Zanger, Ulrich M. Zanger, Ulrich M. Zechel, Christina Zell, Roland Zemp, Ivo Zhang, Suisheng

Zidek, Nadine Zipper, Hubert Zlotnik, Albert zollmann, frank Zywietz, Alexandra

The role of co- and posttranslational modifications in the propagation of scrapie-prions RNA as a cytokine [ribokine]: Metalloregulated extracellular eRNA and bioaptamers, the uncovered missing links for cellular communication and signal transduction in an anticipated RNA world of today. Biomolecular switch from oxidative stress in inflammation to tissue morphogenesis in healing: Fenton-type OH* radical reactions and isoguanosine in metalloregulated RNA bioaptamers of S100-proteins THE EXPRESSION OF ANGIOGENIC GROWTH FACTORS IN HUMAN GRANULOSA CELLS ARE CONTROLLED BY HUMAN CHORION GONADOTROPIN. Natural ligand identification and functional characterization of orphan G protein-coupled receptors Are GTP-analogs really good analogs? Localisation of fluorescently labelled Ras protein in the living cell Involvement of La protein in alternative translation processes in neural progenitor cells Structure of an RNA Involved in Enteroviral Replication Development of an in vitro system for detection and quantitation of urothelial genotoxicity of tobacco smoke-specific constituents Effect of BMP-2 on the migratory and invasive properties of breast cancer cells The intracellular N-terminus of aminopeptidase N/CD13 affects association with caveolae Aminopeptidase N/CD13 is associated with Lubrol rafts Microarray analysis of mucosal gene expression profiles during gastric H. pylori infection Elevated expression level of survivin protein in soft-tissue sarcomas Growth reduction of a xenotransplanted human Soft Tissue Sarcoma by MDM2 Antisense Therapy A new world of tiny RNAs - siRNAs and miRNAs FUNCTIONAL SIGNIFICANCE OF GENETIC POLYMORPHISMS OF HUMAN CYP2B6 ThioTEPA and Clopidogrel are specific mechanism-based inhibitors of human CYP2B6 Putative role of the nuclear factor NCNF in neuronal differentiation Structure of an RNA Involved in Enteroviral Replication Circularly permuted proteins with alternative folding possibilities PHYSICAL INTERACTION BETWEEN NUCLEAR DNA HELICASE II AND WRN HELICASE STIMULATES WRN´S EXONUCLEASE ACTIVITY Comparison of the effects of the P2Y12 ADP-receptor antagonist (AR-C69931) and the GPIIb/IIIa antagonist GR 144053F on platelet-leukocyte conjugate formation. Investigations on the binding of SYBR Green I to double-stranded (ds)DNA A role for Chemokine Receptors in Cancer Metastasis? The cardiac transcription factor Nkx2-5 regulates the alternative promoters of the endothelin-converting enzyme-1 gene in H9c2 cardiomyoblasts G-Protein mediated signalling: Studies in conditional Galpha knockouts