DISCUSSIONS OF THE FARADAY SOCIETY No. 20, 1955 THE PHYSICAL CHEMISTRY OF ENZYMES THE FARADAY SOCIETY Agents f o r the Society's Publications : The Aberdeen University Press Ltd. 6 Upper Kirkgate A berdeenThe Faraday Society reserves the copyright of all Communications published in the ‘‘ Discussions ” PUBLISHED 1956 PRINTED IN GREAT BRITAIN AT THE UNIVERSITY PRESS A B E R D E E NA GENERAL DISCUSSION ON THE PHYSICAL CHEMISTRY OF ENZYMES A GENERAL DISCUSSION on the Physical Chemistry of Enzymes was held in the University Laboratory of Physiology, Parks Road, Oxford (by kind permission of the Vice-Chancellor) on the 10, 11 and 12th August, 1955. The President, Prof. R. G. W. Norrish, F.R.S., was in the Chair and over 250 members and visitors were present. Among the distinguished members and guests welcomed by the President Dr.S. J. Adelstein (Boston, Mass.), Dr. R. L. Baldwin (Wisconsin), Dr. H. J. Berg (Eindhoven), Dr. and Mrs. F. Bergmann (Jerusalem), Dr. S. A. Bernhard (Bethesda, Md.), Dr. and Mrs. A. Bloch (Brunswick, N.J.), Dr. J. J. Blum (Bethesda, Md.), Prof. C. J. F. Bottcher (Leiden), Dr. D. J. Botts (Bethesda, Md.), Prof. and Mrs. P. D. Boyer (Minnesota), Dr. W. Brackman (Amsterdam), Prof. and Mrs. Britton Chance (Pennsylvania), Dr. J. A. Cohen (Holland), Prof. Harold Edelhoch (Kansas), Dr. and Mrs. Eric Ellenbogen (Pittsburgh), Dr. F. G. M. Elliott (Belgium), Dr. F. L. Feitelson (Jerusalem), Dr. Seymour L. Friess (Bethesda, Md.), Dr. Maria Fuld (Pittsburgh), Dr. Alfred Gierer (Tubingen), Mr. L. Ginsaar (Holland), Prof.and Mrs. S. F. Gomes da Costa (Lisbon),Prof. B. C. Guha (Calcutta), Dr. W. F. Harrington (Copenhagen), Prof. F. Haurowitz (Indiana), Dr. Edith Heilbronn (Sweden), Dr. F. L. Hoch, Dr. 0. Hoffman-Ostenhof (Vienna), Dr. B. J. Jandorf (Army Chemical Centre, Md.), Dr. G. M. Kellerman (Australia), Dr. D. Kertesz (Tunis), Dr. T. E. King (Oregon), Prof. J. G. Kirkwood (New Haven, Conn.), Dr. and Mrs. D. E. Koshland, Jr. (New York), Prof. K. J. Laidler (Ottawa), Dr. F. Lindner (Frankfurt), Dr. and Mrs. J. W. Lorimer (Leiden), Prof. and Mrs. J. Murray Luck (Stanford), Prof. R. Lumry (Minnesota), Dr. J. Lundin (Sweden), Dr. A. K. Mills (Dublin), Dr. Manuel Morales (Bethesda, Md.), Prof. and Mrs. H. Neurath (Seattle), Dr. S. 0. Nielsen (Copenhagen), Dr. Z. Nikuni (Japan), Miss Ursula Olpp (Germany), Mr.A. J. J. Ooms (Holland), Dr. R. A. Oosterbaan (Holland), Dr. Sven Paleus (Stockholm), Prof. S. R. Palit (Calcutta), Miss A. M. Paolucci (Rome), Dr. J.-F. Pechere (Brussels), Dr. D. Perlmann (Princeton, N.J.), Dr. Gertrude Perlmann (New York), Dr. D. Peters (Hamburg), Dr. Renzo Rendi (Rome), Dr. and Mrs. D. Rittenberg (New York), Dr. and Mrs. L. Robert (Paris), Prof. R. A. Robinson (Singapore), Ir. M. Sangster (Amsterdam), Prof. R. L. Scott (California), Dr. and Mrs. I. W. Sizer (Cambridge, Mass.), Prof. E. C. Slater (Amsterdam), Dr. L. A. Sluyterman (Eindhoven), Prof. E. L. Smith (Utah), Dr. Bo Sorbo (Stockholm), Prof. and Mrs. M. A. Stahmann (Wisconsin), Mr. B. R. Stein (Gottingen), Dr. G. Stein (Jerusalem), Dr. R. F.Steiner (Bethesda, Md.), Prof. and Mrs. J. M. Sturtevant (New Haven, Conn.), Dr. P. Talalay (Chicago), Prof. and Mrs. H. Theorell (Stockholm), Dr. Fred Vaslow (Copenhagen), Dr. Birgit Vennesland (Chicago), Dr. Mary C . Voorneman (The Hague), Dr. and Mrs. J. N. Walop (Holland), Miss M. G. P. J. Warringa (Holland), Dr. and Mrs. Wetlaufer (Copenhagen), Dr. M. B. Williamson (Chicago), Prof. I. B. Wilson (New York). were the following : 3A GENERAL DISCUSSION ON THE PHYSICAL CHEMISTRY OF ENZYMES A GENERAL DISCUSSION on the Physical Chemistry of Enzymes was held in the University Laboratory of Physiology, Parks Road, Oxford (by kind permission of the Vice-Chancellor) on the 10, 11 and 12th August, 1955. The President, Prof. R. G. W. Norrish, F.R.S., was in the Chair and over 250 members and visitors were present.Among the distinguished members and guests welcomed by the President Dr. S. J. Adelstein (Boston, Mass.), Dr. R. L. Baldwin (Wisconsin), Dr. H. J. Berg (Eindhoven), Dr. and Mrs. F. Bergmann (Jerusalem), Dr. S. A. Bernhard (Bethesda, Md.), Dr. and Mrs. A. Bloch (Brunswick, N.J.), Dr. J. J. Blum (Bethesda, Md.), Prof. C. J. F. Bottcher (Leiden), Dr. D. J. Botts (Bethesda, Md.), Prof. and Mrs. P. D. Boyer (Minnesota), Dr. W. Brackman (Amsterdam), Prof. and Mrs. Britton Chance (Pennsylvania), Dr. J. A. Cohen (Holland), Prof. Harold Edelhoch (Kansas), Dr. and Mrs. Eric Ellenbogen (Pittsburgh), Dr. F. G. M. Elliott (Belgium), Dr. F. L. Feitelson (Jerusalem), Dr. Seymour L. Friess (Bethesda, Md.), Dr. Maria Fuld (Pittsburgh), Dr.Alfred Gierer (Tubingen), Mr. L. Ginsaar (Holland), Prof. and Mrs. S. F. Gomes da Costa (Lisbon),Prof. B. C. Guha (Calcutta), Dr. W. F. Harrington (Copenhagen), Prof. F. Haurowitz (Indiana), Dr. Edith Heilbronn (Sweden), Dr. F. L. Hoch, Dr. 0. Hoffman-Ostenhof (Vienna), Dr. B. J. Jandorf (Army Chemical Centre, Md.), Dr. G. M. Kellerman (Australia), Dr. D. Kertesz (Tunis), Dr. T. E. King (Oregon), Prof. J. G. Kirkwood (New Haven, Conn.), Dr. and Mrs. D. E. Koshland, Jr. (New York), Prof. K. J. Laidler (Ottawa), Dr. F. Lindner (Frankfurt), Dr. and Mrs. J. W. Lorimer (Leiden), Prof. and Mrs. J. Murray Luck (Stanford), Prof. R. Lumry (Minnesota), Dr. J. Lundin (Sweden), Dr. A. K. Mills (Dublin), Dr. Manuel Morales (Bethesda, Md.), Prof. and Mrs. H. Neurath (Seattle), Dr.S. 0. Nielsen (Copenhagen), Dr. Z. Nikuni (Japan), Miss Ursula Olpp (Germany), Mr. A. J. J. Ooms (Holland), Dr. R. A. Oosterbaan (Holland), Dr. Sven Paleus (Stockholm), Prof. S. R. Palit (Calcutta), Miss A. M. Paolucci (Rome), Dr. J.-F. Pechere (Brussels), Dr. D. Perlmann (Princeton, N.J.), Dr. Gertrude Perlmann (New York), Dr. D. Peters (Hamburg), Dr. Renzo Rendi (Rome), Dr. and Mrs. D. Rittenberg (New York), Dr. and Mrs. L. Robert (Paris), Prof. R. A. Robinson (Singapore), Ir. M. Sangster (Amsterdam), Prof. R. L. Scott (California), Dr. and Mrs. I. W. Sizer (Cambridge, Mass.), Prof. E. C. Slater (Amsterdam), Dr. L. A. Sluyterman (Eindhoven), Prof. E. L. Smith (Utah), Dr. Bo Sorbo (Stockholm), Prof. and Mrs. M. A. Stahmann (Wisconsin), Mr. B. R.Stein (Gottingen), Dr. G. Stein (Jerusalem), Dr. R. F. Steiner (Bethesda, Md.), Prof. and Mrs. J. M. Sturtevant (New Haven, Conn.), Dr. P. Talalay (Chicago), Prof. and Mrs. H. Theorell (Stockholm), Dr. Fred Vaslow (Copenhagen), Dr. Birgit Vennesland (Chicago), Dr. Mary C . Voorneman (The Hague), Dr. and Mrs. J. N. Walop (Holland), Miss M. G. P. J. Warringa (Holland), Dr. and Mrs. Wetlaufer (Copenhagen), Dr. M. B. Williamson (Chicago), Prof. I. B. Wilson (New York). were the following : 3CONTENTS PAGE General Introduction. By M. Dixon . . 9 I. CHARACTERIZATION AND PHYSICAL PROPERTIES- Transport Processes and the Heterogeneity of Proteins. By R. L. Baldwin, L. J. Gosting, J. W. Williams and R. A. Alberty . . 13 Certain Physical Properties of Chymotrypsin and Chymotrypsinogen using the Depolarization of Fluorescence Technique.By V. Massey, W. F. Harrington and B. S. Hartley . . 24 Mechanism of Activation of Trypsinogen and Chymotrypsinogen. By H. Neurath and W. J. Dreyer . . 32 Activity of Catalase-Lipid Complexes at Oil/Water Interfaces. By M. J. Fraser, J. G. Kaplan and J. H. Schulman . . 44 Activation and Inactivation of Milk Xanthineoxidase by Physico- chemical Means. By L. Robert and J. Polonovski . . 54 GENERAL DIscussIoN.-Dr. D. B. Wetlaufer, Prof. H. Neurath, Dr. Hartley, Dr. V. Massey, Dr. G. A. Gilbert, Prof. F. Haurowitz, Prof. J. G. Kaplan, Dr. L. Robert, Dr. R. K. Morton, Dr. Tsoo E. King, Dr. V. H. Cheldelin, Prof. D. D. Eley . 65 11. KINETICS AND MECHANISMS- The Influence of Fluctuations in Protein Charge and Charge Con- figuration on the Rates of Enzymatic Reactions. By J.G. Kirkwood 78 Some Kinetic and Mechanistic Aspects of Hydrolytic Enzyme Action. By K. J. Laidler . . 83 Some Chemical and Kinetic Studies of Crystalline Papain. By E. L. Smith, B. J. Finkle and A. Stockell . . 96 The Chemical Kinetics of Some Reactions Catalyzed by Pancreatic Carboxypeptidase. By R. Lumry and E. L. Smith . . 105 The Chemical Structure of the Reactive Group of Esterases. By J. A. Cohen, R. A. Oosterbaan, M. G. P. Warringa and H. S. Jansz . 114 Promotion of Acetylcholinesterase Activity by the Anionic Site. By I. B. Wilson . 119 Fine Structure of the Active Surface of Cholinesterases and the 5 Mechanism of Enzymatic Ester Hydrolysis. By F. Bergmann . 1266 CONTENTS The Mechanism of Reaction Between Esterases and Phosphorus- Containing Anti-Esterases. By B.J. Jandorf, H. 0. Michel, N. K. Schaffer, R. Egan and W. H. Summerson. . PAGE Isotopic Exchange Criteria for Enzyme Mechanisms. By D. E. Koshland, Jr. . The Group-Transfer Activity of Certain Hydrolytic Enzymes. By R. K. Morton Enzyme Action at a Distance. An Examination of its Kinetics and Physical Possibilities. By G. Weber . . Activation and Inhibition of Enzymes. By A. G. Ogston . Steps in the Formation and Decomposition of Some Enzyme-Substrate Complexes. By H. Gutfreund . Hydrogen-Activating Enzymes of Bacteria. By A. Couper, D. D. Eley and A. Hayward . Interaction of Hydrogenase with Hydrogen. By D. Rittenberg and A. I. Krasna . The Action of Fluorocitric Acid on Aconitase. By R.A. Peters . The Kinetics of Haemoglobin and Haem Compounds as Models for . Enzyme Action. By Q. H. Gibson and F. J. W. Roughton Intracellular Reaction Kinetics. By B. Chance . The Haem-Linked Ionizing Group in Myoglobin. By P. George and G. I. H. Hanania . On the Interaction Between Coenzymes and Enzymes. By H. Theorell Calculation of the Rate Constants of the Reaction Between an Enzyme and its Substrate from the Overall Kinetics of the Reaction Catalyzed by the Enzyme. By E. C. Slater , Some Applications of Deuterium to the Study of Enzyme Mechanisms. By B. Vennesland . The Hydrogen Ion in the Enzymatic Oxidation and Reduction of DPN. By I. W. Sizer and A. Gierer . GENERAL D~scussIoN.-Prof. K. J. Laidler, Prof. J. M. Sturtevant, Dr. D. J. R. Laurence, Dr. H. Gutfreund, Prof. H. Neurath, Prof. R. Lumry, Dr. B. R. Rabin, Prof. E. L. Smith, Dr. R. J. P. Williams and Dr. B. L. Vallee, Dr. L. Robert, Dr. J. J. Blum, Dr. S . A. Bernhard, Dr. F. Bergmann, Dr. P. D. Boyer, Dr. D. R. Davies and Dr. A. L. Green, Dr. R. K. Morton, Dr. D. E. Koshland, Jr., Mr. P. A. T. Swoboda, Dr. 0. Hoffmann-Ostenhof, Dr. D. D. Eley, Dr. D. Bunn, Prof. J. A. V. Butler, Dr. M. Sangster, Dr. A. Klug, and Prof. F. J. W. Roughton, Mr. J. G. Beetlestone and Dr. A. Couper, Prof. H. N. Rydon, Dr. H. Kacser, Prof. F. Haurowitz, 134 142 149 156 161 167 174 185 189 195 205 216 224 23 1 240 248CONTENTS 7 PAGE Dr. K. Dalziel, Dr. P. George, Prof. Q. H. Gibson, Mr. J. S. Griffith, Dr. G. I. H. Hanania, Dr. S . J. Adelstein and Dr. J. A. Olson, Dr. F. L. Hoch, Mr. W. E. Trevelyan, Dr. M. Dixon, Dr. L. L. Ingraham, Dr. E. C. Slater, Dr. B. Vennesland, Dr. P. Talalay, Dr. I. W. Sizer and Dr. A. Gierer . . 254 Author Index . . 317