34. Colloquium der Gesellschaft fOr Biologische Chemie 14. -16. April 1983 in Mosbach/Baden Biological Oxidations Edited by H. Sund and V. Ullrich With 119 Figures Springer-Verlag Berlin Heidelberg New York Tokyo 1983 Professor Dr. HORST SUND Professor Dr. VOLKER ULLRICH Fakultat fOr Biologie der Universitat Konstanz Postfach 5560 0-7750 Konstanz, FRG ISBN-13: 978-3-642-69469-1 e-ISBN-13: 978-3-642-69467-7 001: 10.1007/978-3-642-69467-7 This work is subject to copyright. All rights are reserved, whether the whole or part of the material is concerned, specifically those of translation, reprinting, re-use of illustrations, broadcasting, reproduction by photocopying machine or similar means, and storage in data banks. Under § 54 of the German Copyright Law, where copies are made for other than private use, a fee is payable to ''Verwertungsgesellschaft Wort", Munich. © by Springer-Verlag Berlin Heidelberg 1983 Softcover reprint of the hardcover 1st edition 1983 The use of registered names, trademarks etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. 2131f313G-543210 Preface This volume contains the lectures and discussions of the 34th Mosbach Colloquium, organized by the Gesellschaft fur Biologische Chemie in commemoration of otto Warburg's 100th anniversary. Following warburg's main scientific interests the topics of the contributions were related to the hydrogen-transferring enzymes as well as to the oxygen-reducing and oxygen-activating enzymes. Due to the detailed knowledge on the molecular structure of NAD- and flavin-dependent dehydrogenases, a better understanding of enzyme catalysis and especially enzyme evolu tion has emerged. In the field of oxidases and oxygenases we begin to understand the complex biochemistry of oxygen and realize that binding, reduction, and activation of this molecule have common underlying prin ciples. Even new metabolic pathways of oxygenases, such as lipoxygen ases or cyclooxygenase, have recently been discovered and promise new innovations in cell regulation and medicine. Thanks to the cooperation of all authors, a well-balanced mixture of basic information and progress report was provided. We especially ap preciate Professor Bucher's charming, thoughtful, and honest contri bution on Otto Warburg as a skilful scientist and colorful personality. The editors are indebted to the sponsoring organizations, the Deutsche Forschungsgemeinschaft and several industrial companies, and especial ly acknowledge the competent help of Mrs. H. Allen in the editorial work. October 1983 H. SUND V. ULLRICH Contents Otto Warburg: A Personal Recollection Th. BUcher (With 15 Figures) ••..•••••.••••••••••..••••..•••••••• NAD-Dependent Dehydrogenases structure-Function Relationships of NAD-Dependent Dehydrogenases M.G. Rossmann (With 8 Figures) •••••••••••••••••••••••••••••••••• 33 Binding of Coenzyme Analogs to NAD-Dependent Dehydrogenases J .F. Biellmann (With 9 Figures) •.•.•••..•••.••.•••••••.•...•..•. 55 What NAD-Dependent Dehydrogenases Teach Us About the Folding and Association of Oligomeric Proteins R. Jaenicke and R. Rudolph (With 7 Figures) ..•.••....•..••.•..•• 62 Flavoproteins Flavoproteins of Known Three-Dimensional Structure R.H. Schirmer and G.E. Schulz (With 9 Figures) •..••....•...••.•• 93 The Mechanism of Action of Flavoprotein-Catalyzed Reactions v. Massey and S. Ghisla (With 25 Schemes and 2 Figures) ....•.•••• 114 Mechanistic Studies on Mercuric Ion Reductase and Cyclohexanone Oxygenase: Pharmacologic and Toxicologic Aspects C. Walsh, B. Branchaud, B. Fox, and J. Latham •.••..•••..••.••••. 140 Dioxygen Binding and Reduction Oxygenases and Oxidases: Hypothesis H.S. Mason (With 12 Figures) .••.••••...•..•••.••.••...••..•••.•• 151 Reversible Dioxygen Binding K. Gersonde (With 13 Figures) 170 Cytochrome c Oxidase and Related Enzymes B. G. Malmstrom (With 4 Figures) ..•....••..•••.•..••..••.....•... 189 VIII Oxygenases Lipoxygenases from Plant and Animal Origin J.F.G. Vliegenthart, G.A. Veldink, J. Verhagen, and S. Slappendel (With 8 Schemes and 8 Figures) .•..•...•••.•..••••.•......•.•..•.• 203 Physiological Reactions of Arachidonic Acid Oxygenation Products R. J. Flower ...•...••..•...•....•••..•••••.•••••......•.•......•• 224 Bacterial Cytochrome P-450 Monooxygenases G.C. Wagner (With 3 Figures) •.•••••.••••.••••..•.••.••..•••••.•• 234 Reactive Intermediates Derived from Cytochrome P-450 Monooxygenases D. Mansuy ••••••••••••••.•••••••••••.•••.••.••..•.•••••.••...•... 240 Integrated Systems Regulation of Glycolysis B. Hess, A. Boiteux, and D. Kuschmitz (With 15 Figures) •••••.••. 249 Regulation of ATP Synthesis in Mitochondria M. Klingenberg (With 5 Figures) •••••••....•••...•..•.•.•........ 267 Oxygenation Pathways in Bacteria R. Muller and F. Lingens (With 11 Figures) •.••...••..........••• 278 Oxygen Radicals and Hydroperoxides in Mammalian Organs: Aspects of Redox Cycling and Hydrogen Peroxide Metabolism E. Cadenas, R. Brigelius, Th. Akerboom, and H. Sies (With 3 Figures) ••..•..••••.•••.•...•••.....•..••.•..••.•.•..... 288 Subj ect Index •.•.•••.....•........•............................. 311 Contributors You will find the addresses at the beginning of the respective contribution Akerboom, Th. 288 Lingens, F. 278 Biellmann, J.F. 55 Malmstrom, B.G. 189 Boiteux, A. 249 Mansuy, D. 240 Branchaud, B. 140 Mason, H. S . 1 51 Brigelius, R. 288 Massey, V. 11 4 BUcher, Th. 1 MUller, R. 278 Cadenas, E. 288 Rossmann, M.G. 33 Flower, R.J. 224 Rudolph, R. 62 Fox, B. 140 Schirmer, R.H. 93 Gersonde, K. 170 Schulz, G.E. 93 Ghisla, S. 114 Sies, H. 288 Hess, B. 249 Slappendel, S. 203 Jaenicke, R. 62 Veldink, G.A. 203 Klingenberg, M. 267 Verhagen, J. 203 Kuschmitz, D. 249 Vliegenthart, J.F.G. 203 Latham, J. 140 Wagner, G.C. 234 Walsh, C. 140 Otto Heinrich warburg 3 October 1883 - 1 August 1970 Otto Warburg: A Personal Recollection Th.BOcher1 Pupils as Biographers It is to Hans Adolf Krebs that we owe an authoritative biography of Otto Warburg. Originally written as an obituary for the Royal Society of London [1], it was also published in German [2]. The first written communication by Krebs from Warburg's laboratory dates from 1927 [3]. In the 43 years preceding Warburg's death, there existed a bond be tween the two men profiting in its personal as well as in its profes sional aspect from the stimulating tension of a teacher-pupil relation ship. Krebs had most probably kept a diary with the thoroughness that characterized his style of working. In addition to this, he studied the roots of Warburg's personality and investigated the sources of the publications. After this biography, which in its affectionate sincerity honors both the biographer and his subject, what can be the further purpose and sense of my contribution? Any appraisal of Warburg within the historical scope of natural sci ~nces should be left to the experts. Nevertheless, it so happens that I met Otto Warburg in those very years in which Krebs, due to his emigration and the war, had lost direct contact with the Dahlem Group. The lack of Krebs' personal experience leads to a less vivid represen tation of the decisive period in which Warburg and his coworkers laid the foundations for many of the contributions to this Colloquium. Con sequently it may appear sensible to continue and complete Krebs' at tempt to "let Warburg speak in his own words", to take the risk of tracing features of his personality as remembered by his pupil. On this matter a few words on my own behalf: neither before nor after the years in Warburg's working team did I meet a teacher in whom I was able to confide so unreservedly. The span of time between autumn 1938 until spring 1945, interrupted by 14 months of military service, was decisive for me far beyond its professional aspect. I therefore see little sense in hiding my own person in the following scenes, an en deavor which inCidentally would have been a striking example of what Warburg would have called an Eiertanz.2,3 Physiologisch-Chemisches Institut, GoethestraBe 33, D-8000 Munchen, FRG 2 This lecture was originally given in German ("ilber Otto Warburg in der Erinnerung") and, at the request of the publishers, has been translated into English. I am grateful to Mrs. H. Allen and Mrs. A. Macolister, University of Constance, and to Ms. H. Carr for attempting to put my thoughts into a comprehensible English version 3 Rural folk dance amongst a pattern of fresh eggs, where the dancer tries not to break the shells. Such scenes were frequently depicted by Dutch genre painters, e.g., The Christening Feast by Jan Steen (P111 at the Wallace Collection, London) 34. Colloquium-Mosbach 1983 Biological Oxidations c Springer-Verlag Berlin Heidelberq 1983 2 Application for a Position as a Doctoral Candidate The intellectual Berlin of the early thirties was characterized by the most lively participation in the development of the arts and sci ences. The concentration of creative authorities was unparalleled. In the socially different worlds of pre- and post-war Berlin, turning to progress in the same way as assimilating it, Otto Warburg became well-known and influential. Where formerly the principle of unity of research and teaching had taken shape, he represented a new type of "pure researcher" that fascinated the young. The public of that time took less notice of Nobel prizes than today, and turned their atten tion far more to the lecture evenings of the scientific societies, above all to those of the Kaiser-Wilhelm-Gesellschaft which was even honoured by the visit of the Reichsprasident Ebert in his time. In 1926, Warburg lectured to the medical corps on the cancer problem [4], in 1928 in the Harnack Haus on the action spectrum of the respi ratoryenzyme [5]. Fascinating discoveries were quickly discussed everywhere, and were soon incorporated in the teaching syllabus of the grammar schools, at least in those of Berlin's western suburbs in which the Kaiser-Wilhelm Institutes were situated. I am unable to say exactly what motivated me even during my last school years at Berlin-Steglitz' Paulsen Realgymnasium, and later dur ing my chemical studies, to work toward an apprenticeship with Otto Warburg. It was most likely the influence of our esteemed biology teacher. At his suggestion, Albert Reid - a schoolfellow 4 years my senior - had written his Ph.D. under the supervision of Warburg. At any rate, immediately after passing my final examinations in chemistry, I made enquiries as to how I could get in contact with him. In general, Warburg not only had the reputation of being a genius, but also of being arrogant. I therefore chose the shortest, but also the least compromising way, by phoning the institute to ask for acceptance as a doctoral candidate. Without more ado, with an appointment for 6 o'clock the same after noon, I met Otto Warburg in the beautiful, oval-shaped library of his institute. I saw him for the first time. He was of rather slight sta ture, but with broad shoulders. The sleeves of his elegant woollen waistcoat were slightly frayed. Gray tweed trousers and carefully polished Scottish shoes completed the picture of a nobleman of the British School. I repeated my request in a few sentences. Warburg looked directly into my face, his mouth closed. His look did not frighten, on the contrary, but it was penetrating, as I was soon to learn. Without going into lengthy questions concerning my person, Warburg decided that the consent of the Head of the Department should be obtained. By this, the requested opportunity was probably granted in principle. He then said, unexpectedly, establishing with incomparable authority the teacher-pupil relationship: "I presume you do not know what we are working on ••• I will get you a publication."