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ACADEMIC PRESS RAPID MANUSCRIPT REPRODUCTION
Johnson Research Foundation Colloquia
Energy-Linked Functions of Mitochondria Edited by Britton Chance 1963
Rapid Mixing and Sampiing Techniques in Biochemistry Edited by Britton Chance, Quentin H. Gibson, Rudolph H. Eisenhardt, K. Karl Lonberg-Holm 1964
Control of Energy Metabolism Edited by Britton Chance, Ronald W. Estabrook, John R. Williamson 1965
Hernes and Hemoproteins Edited by Britton Chance, Ronald W. Estabrook, Takashi Yonetani 1966
Probes of Structure and Function of Macromolecules and Membranes Volume I Probes and Membrane Function Edited by Britton Chance. Chuan-pu Lee. J. Kent Blasie 1971
Probes of Structure and Function of Macromolecules and Membranes Volume II Probes of Enzymes and Hemoproteins Edited by Britton Chance, Takashi Yonetani, Albert S. Mildvan 1971
Biological and Biochemica! Oscillators Edited by Britton Chance, E. Kendall Pye. Amal K. Ghosh, Benno Hess 1973
Alchohol and Aldehyde Metabolizmg Systems Edited by Ronald G. Thurman. Takashi Yonetani, John R. Williamson. Britton Chance 1974
Alcohol and Aldehyde Metabolizing Systems Volume II Enzymology and Subcellular Organelles Edited by Ronald G. Thurman, John R. Williamson, Henry R. Drott, Britton Chance 1977
Alcohol and Aldehyde Metabolizing Systems Volume III Intermediary Metabolism and Neurochemistry Edited by Ronald G. Thurman, John R. Williamson, Henry R. Drott. Britton Chance 1977
Frontiers of Biological Energetics Volume I Electrons to Tissues Edited by P. Leslie Dutton, Jack S. Leigh, Antonio Scarpa 1978
Frontiers of Biological Energetics Volume II Electrons to Tissues Edited by P. Leslie Dutton, Jack S. Leigh, Antonio Scarpa 1978
Frontiers of
Biological Energetics Volume I: Electrons to Tissues
Edited by P. Leslie Dutton Jack S. Leigh
Antonio Scarpa
Johnson Research Foundation and
Department of Biochemistry and Biophysics University of Pennsylvania Philadelphia, Pennsylvania
A C A D E M I C PRESS New York San Francisco London
A Subsidiär}' of H a r c o u r t Bruce Jovanovich, Publishers
Contents
Contributors for Volumes I and II P*reface
P a r t 1 E l e c t r o c h e m i c a l I n t e r a c t i o n s
Vectorial Electron, Hydrogen, Proton, and Oxyanion Conduction in Chemiosmotic Reaction Mechanisms Peter M i t c h e l l
Energetic Advantage of Ion Countertransport in Chemiosmotic Conversion G r e g o r i o Weber
Tunneling Processes in Bacteriophotosynthesis and Vision P . M . Rentzepis
On the Efficiency of Electron Transfer Reactions in Proteins A . W a r s h e l a n d R. Weiss
Kinetics of Photochemical Electron Transfer Reactions i n V i v o and in V i t r o W. W. P a r s o n , C. C. Schenck, R. E . B l a n k e n s h i p , D . H o l t e n , M . W. W i n d s o r , a n d C. V. Shank
Structural Organization of the Redox Groups in a Bacterial Photosynthetic Reaction Center Complex D . M . T i e d e , J . S. L e i g h , a n d P . L . D u t t o n
Geometrical Relationships between the Cytochrome c-Cytochrome Oxidase Mitochondrial Redox Pairs Jane V a n d e r k o o i , J . S. L e i g h , C h a r l e s S. O w e n , Peter G l a t z , a n d H a y w o o d B l u m
Excited and Ionic States of Dimeric Chlorophyll Derivatives. Biomimetic Modelling of the Primary Events of Photosynthesis M i c h a e l R. W a s i e l e w s k i
vii
Covalently-Linked Porphyrin Quinone Complexes as R C Models Josephine L . Y. K o n g a n d P a u l A . L o a c h
Protein Dynamics, Potential Regulation, and Redox Coupled Conformational Changes in Cytochromes c F . R. Salemme
Electron Transfer by Cytochromes: Mechanisms and Problems M i c h a e l A . C u s a n o v i c h
Cytochromes c2—An Evolutionary Family R i c h a r d E . D i c k e r s o n a n d T s u n e h i r o T a k a n o
Evolutionary Changes of the Herne C Electronic Structure in Cytochromes c K u r t Wüthrich, R e g u l a M . K e l l e r , a n d Sidney L . G o r d o n
Respiratory Proteins of Some Extremely Thermophilic Bacteria James A . F e e , K a r e n L . F i n d l i n g , A l i s o n Lees, a n d T a t s u r o Y o s h i d a
Alternative Cyanide-Insensitive Respiratory Chain in P a r a c o c c u s d e n i t r i f i c a n s M i c h e l e F . H e n r y a n d P a u l e t t e M . V i g n a i s
Specification of and Regulation by the Cytochrome B Region of the Mitochondrial Genome in Yeast H e n r y R. M a h l e r , D e b o r a h K . H e n s o n , a n d D o n a l d H . M i l l e r
Biosynthesis and Intracellular Translocation of Mitochondrial Proteins: Cytochrome c and the Carboxyatractyloside Binding Protein R i c h a r d Z i m m e r m a n n , H a r o l d K o r b , a n d W a l t e r N e u p e r t
Labelling of Complex III with 3 5 S Diazobenzenesulfonate. Orientation of this Electron Transfer Segment in the Mitochondrial Inner Membrane Randy L . B e l l , Jeanne Sweetland, B e r n d L u d w i g , a n d R o d e r i c k A . C a p a l d i
Thermodynamic and EPR Properties of Hydrogen Carriers in the Succinate-Cyt. C Reductase Segment of the Respiratory Chain T o m o k o O h n i s h i , J . C. S a l e r n o , T. M a i d a , C. A . Yu, S. N a g a o k a , a n d T. E . K i n g
Contents
Mitochondrial Ubiquinone Proteins Tsoo E . K i n g , L . Yu, S. N a g a o k a , W. R. W i d g e r , a n d C . A . Yu 174
Thermodynamic and Functional Heterogeneity among the Ubiquinones of Rhodopseudomonas s p h a e r o i d e s K e n - I c h i r o T a k a m i y a , Roger C. P r i n c e , a n d P . L e s l i e D u t t o n 183
Electron and Proton Transfer in Succinate-Cytochrome c Reductase Segment of the Respiratory Chain J . C. S a l e r n o , T. M a i d a , H . B l u m , a n d T. O h n i s h i 191
A Single Ubiquinone Plays a Central Role in Electron Flow Through the Ubiquinone-Cytochromes b-c2
Oxidoreductase Roger C. P r i n c e , W i l l e m H . v a n den B e r g , K e n - I c h i r o T a k a m i y a , C. L i n d s a y B a s h f o r d , a n d P . L e s l i e D u t t o n 201
Effects of Ethanol and Salicylhydroxamic Acid on the Interactions of the Ubisemiquinone Pair with Centre S-3 in Plant Mitochondria A n t h o n y L . M o o r e , D a v i d O. H a l l , a n d H e i n z Rupp 210
Involvement of the 4 4 Protein" in Redox-Coupled Protonation Events of the Quinone Acceptor-Complex in Bacterial Photosynthetic Reaction Centers C. A . W r a i g h t 218
Pigment Interaction and Picosecond Electron Transfer in Bacterial Reaction Centers V. A . Shuvalov 227
Recent Developments on the "Primary" Electron Acceptors in Photosystem I B a c o n K e , V l a d i m i r A . Shuvalov, a n d E d D o l a n 234
Redox Potential Dependence of Electron Transport and Variable Fluorescence in Photosystem I M . C . W . Evans, P . H e a t h c o t e , A . Telf er, a n d J . B a r b e r 241
Flash-Induced Volume Changes in Purple Membrane Suspensions D o n a l d R. O r t a n d W i l l i a m W. P a r s o n 249
Establishments of Electrochemical Gradients: General Views and Experiments on Purple Membrane Benno Hess a n d D i e t r i c h K u s c h m i t z 257
X Contents
Site Specific Interaction of Protons Liberated from Photosystem II Oxidation with a Hydrophobie Membrane Component of the Chloroplast Membrane L . J . P r o c h a s k a a n d R. A . D i l l e y 265
Calibration of Flash Induced pH Changes inside Thylakoids and Kinetic Resolution of Proton Ejection and Consumption Wolfgang J u n g e , A l l i s o n M c G e e r , a n d Winfried Ausländer 275
P a r t 2 E l e c t r o n s , P r o t o n s , a n d E n e r g y
Formation of the Proton Gradient Across the Chloroplast Thylakoid Membrane in Relation to A T P Synthesis W. 5. C h o w , S. W. T h o r n e , a n d N . K . B o a r d m a n 287
The Molecular Mechanism of Light Activated Proton Transport in Purple Membranes of H a l o b a c t e r i u m h a l o b i u m A a r o n L e w i s 297
Coupling of Electron Transfer and Proton Translocation in Purple Photosynthetic Bacteria H i r o y u k i A r a t a a n d M i t s u o N i s h i m u r a 307
Contributions from Both Electron Transport and Ion Transport to the Decay of the Carotenoid Shift After Flash-Activation of Chromatophores J . B . J a c k s o n , J . A . G r e e n r o d , N . K . P a c k h a m , a n d K . M . Petty 316
Inhibition of Electron Transport in RPS. Capsulata by a Ubiquinone Analogue J o h n R. B o w y e r a n d A n t h o n y R. Crofts 326
Vectorial Oxidoreductions: The Ferrous lron Oxidase Complex of T h i o b a c i l l u s f e r r o o x i d a n s and the Nitrate Reductase Complex of E s c h e r i c h i a c o l i W. J o h n I n g l e d e w , J o h n C. C o x , R o b e r t W. Jones, a n d Peter B . G a r l a n d 334
Electron Translocating Function of Cytochrome Oxidase Peter M i t c h e l l a n d Jennifer M o y l e 342
Generation of Electrochemical Proton Gradient by Mitochondrial Cytochrome c Oxidase. Clarification of Some Controversial Experiments Märten Wikströ'm a n d K l a a s K r a b 351
Control Mechanisms for Proton Conduction in the Mitochondrial H + -ATPase A n g e l a P a n s i n i , F . G u e r r i e r i , a n d S. P a p a 359
Contents x i
The Mechanism of Transmembrane A / X H + Generation by Cytochrome c Oxidase S. P a p a , F . G u e r r i e r i , M , L o r u s s o , F . C a p u a n o , G. I z z o , a n d D . Boffoli 367
The Three Proton Pumps of the Mitochondrial Respiratory Chain G. F . Azzone, T. Pozzan, F . D i V i r g i l i o , a n d V. M i c o n i 375
Proton Stoichiometry of Mitochondrial Electron Transport, A T P Hydrolysis, and ATP-Dependent Reverse Electron Flow A l b e r t L . L e h n i n g e r , B a l t a z a r Reynafarje a n d Adolf A l e x a n d r e 384
Protonmotive Stoichiometry of Redox and ATPase Systems Peter M i t c h e l l , J e n n i f e r M o y l e , a n d Roy M i t c h e l l 394
Membrane Potential, Phase Transitions, and Coupling in Mitochondria H a g a i R o t t e n b e r g 403
Further Studies on the Membrane Potential of Giant Mitochondria Using Microelectrodes C h a r l e s B o w m a n , B r u c e L . Maloff, a n d H e n r y Tedeschi 413
Can Energy Coupling Occur in the Mitochondrial Membrane in the Absence of Transmembrane Gradients? D a n a M . Scott a n d B a y a r d T. Storey 422
The Energy Balance of Oxidative Phosphorylation K . v a n D a m , R. P . Casey, R. v a n der M e e r , A . K . G r o e n , a n d H . V. Westerhoff 430
Effects of Anions on the Reaction between Cytochrome c and Cytochrome c Oxidase N e i l Osheroff, W. H . K o p p e n o l , a n d E . M a r g o l i a s h 439
Proton Translocating ATPase Subunit Structure and Pump, Gate, and Channel Activities N o b u h i t o Sone, Masasuke Y o s h i d a , H a j i m e H i r a t a , a n d Yasuo K a g a x v a 450
ATP-Synthesis Induced by a pH-Gradient Imposed Across a Collagen Film Bearing an ATPase-ATP Synthase C a t h e r i n e G o d i n o t , B r u n o B l a n c h y , P i e r r e R. C o u l e t , a n d D a n i e l e C. G a u t h e r o n 459
Site Directed Modifications of B F r A T P a s e From E . c o l i . Comparison of Binding Properties of Bacterial and Mitochondrial Fj with Respect to Aurovertin, D C C D , and E E D Q M i c h e l S a t r e , R i c h a r d P o u g e o i s , J o e l L u n a r d i , A n n e - C h r i s t i n e D i a n o u s , G e r a r d K l e i n , M i r e i l l e Bof, a n d P i e r r e V. V i g n a i s
Solubilization of Coupling Factor 1 From Chloroplast Thylakoids Alters Its Interactions with Nucleotides R i c h a r d E . M c C a r t y , R o n a l d P . M a g n u s s o n , a n d R a y m o n d W o n g
3 '-Esterified Adenine Nucleotides as Extrinsic Probes of the Energy Coupling Device Günter Schäfer
New Results Reveal Subunit Catalytic Cooperativity in F {
ATPase R. Lee H u t t o n , D a v i d D . H a c k n e y , a n d P a u l D . B o y e r
Reconstitution of Oligomycin- and Dicyclohexylcarbodiimide-Sensitive Mitochondrial ATPase from Isolated Components B i r g i t t a N o r l i n g , E l z b i e t a G l a s e r , a n d L a r s E r n s t e r
Complex V: Composition and Molecular and Enzymic Properties Yves M . G a l a n t e , L u c i a n o F r i g e r i , a n d Youssef Hatefi
Biochemical Characterization of the Electrogenic Proton Pump of the Neurospora Plasma Membrane B a r r y J . B o w m a n , F r a n c i s B l a s c o , a n d C a r o l y n W. S l a y m a n
Measurement of the Protonmotive Force in Amine Containing Subcellular Organelles R. G. J o h n s o n , A . S c a r p a , a n d L . Salganicoff
Enzymic Modification of Gastric Transport ATPase G e o r g e Sachs, E d d R a b o n , a n d G a e t a n o Saccomani
Chloride Transport in Gastric Cells and Microsomes A n n i c k S o u m a r m o n a n d Efraim Racker
Membrane Potential and C l ~ Transport Properties of Primary Glial Cultures from Rat Brain H . K . K i m e l b e r g , S. B i d d l e c o m e , a n d R. S. B o u r k e
Contents . x i i i
The Interaction of M g + + and pH in Chloroplast Processes G e o r g e H o c h a n d S a l i l Bose 573
P a r t 3 New I n s t r u m e n t a l A p p r o a c h e s t o C e l l u l a r B i o p h y s i c s
Electron Spin Echo Spectroscopy and Photosynthesis J . R. N o r r i s , M . C. T h u r n a u e r , M . K . B o w m a n , a n d A . D . Trifunac 581
Use of the Molecular Microprobe to Record Raman Spectra of a Single Mitochondrion and a Fiber of Calf Thymus D N A F r a n A d a r 592
Normal Coordinate Models for Herne Raman Spectra: Uses and Limitations L . R i m a i a n d I . Salmeen 600
Added Precision in 5 7Fe Mossbauer Spectrometry of Proteins W. F . F i l t e r , W. R. D u n h a m , R. M . P o l i c h a r , a n d R. H . Sands 608
Dynamics of the Local Iron Environment by the Selective Excitation Double Mossbauer Technique (SEDM) B . B a l k o , E . V. M i e l c z a r e k , a n d R. L . B e r g e r 617
Exchange Interaction in Spinach Ferredoxin Determined by Electron Paramagnetic Resonance H a y w o o d B l u m , J . S. L e i g h , T o m o k o O h n i s h i , a n d J . C. S a l e r n o 625
Precise EPR Measurements on Small Protein Crystals R. A . L i e b e r m a n , W. R. D u n h a m , J . A . F e e , a n d R. H . Sands 634
How Sensitive is the EPR of Herne to a Perturbed Environment? C. P . S. T a y l o r a n d B . R. Sreenathan 644
X-Ray Absorption Studies of Metalloproteins P . E i s e n b e r g e r , B . M . K i n c a i d , a n d /? . G. S h u l m a n 652
Core Expansion vs. Coming in Porphyrins; Resonance Raman Discrimination between Five- and Six-Coordinate High-Spin Fe"1
Hernes Thomas G. S p i r o , J o h n D . Stong, a n d P a u l Stein 660
Dynamics of Non-Linear Electric Field Effects L e o D e M a e y e r a n d F r e d d y Paulussen 671
Herne Protein Reactions at High Pressure and Low Temperature L a u r a E i s e n s t e i n a n d H a n s F r a u e n f e l d e r
Calorimetric Studies of the Heat of Respiration of Mitochondria I c h i r o M a t s u o k a , T a k a h i d e W a t a n a b e , a n d T a k a o N a k a m u r a
The Measurement of AH and the k k 0 n " Rate Constant of the Reactions of C 0 2 and 2,3-DPG with Deoxyhemoglobin by Thermal Stopped Flow R. L . B e r g e r , B . B a l k o , P . B o w e n , a n d R. P a u l
Studies of Stable Metal-Nucleotide Complexes Interacting with Myosin Subfragment J . F . E c c l e s t o n a n d D . R. Fr entkam
Intracellular Enzyme Activity N o b u t o m o I t a d a , L a r a i n e Peiffer, a n d R o b e r t E . F o r s t e r
E . c o l i Unadenylated Glutamine Synthetase: Elucidation of the Catalytic Cycle and the Role of Some Feedback Inhibitors S. G. Rhee, P . B . C h o c k , a n d E . R. Stadtman
High Voltage Microscopy of Cells and Membranes in the Hydrated State D o n a l d F . P a r s o n s
Mitochondrial and Sarcoplasmic Reticulum Contents i n S i t u : Electron Probe Analysis A . P . S o m l y o , H . Shuman, a n d A . V. Somlyo
Augmentation of Tissue Water Proton Spin-Lattice Relaxation Rates by i n V i v o Addition of Paramagnetic Ions P a u l C. L a u t e r b u r , M . H e l e n a M e n d o n q a D i a s , a n d A n d r e w M . R u d i n
Anomalous X-Ray Scattering Studies for the Determination of the Location of Redox Centers in Membranes—A Feasibility Study James Stamatoff, Peter E i s e n b e r g e r , G e o r g e B r o w n , James Pachence, L e s l i e D u t t o n , J o h n L e i g h , a n d K e n t B l a s i e
Magnetic Resonance Studies of the Mechanism of R N A Polymerase from E . c o l i A . S. M i l d v a n , P . S t e i n , R. K o r e n , a n d B . Bean
Contents
3 1 P N M R of Brain Tissue Under Normal, Hypothermie, and Freeze-Trapped Conditions B . C h a n c e , Y. Nakase, M . B o n d ,
J . S. L e i g h , J r . , a n d G. M c D o n a l d 779
Visualization of Dynamic Spatial Structures in Oscillating Cell Free Extracts of Yeast A r n o l d B o i t e u x a n d Benno Hess 789
Contributors of Volumes I and II
Hiroshi Abe, Osaka University Medical School, First Department of Internal Medicine, Osaka, Japan
Kazuhiko Adachi, Department of Biochemistry and Biophysics, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Fran Adar, Department of Biochemistry and Biophysics, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
N . Akaike, Department of Physiology and Biophysics, University of Texas Medical Branch, Galveston, Texas 77550
Karl E. Akerman, Department of Medical Chemistry, University of Helsinki, Siltavuorenpenger 10, SF 00170 Helsinki 17, Finland
Adolfo Alexandre, Johns Hopkins University, School of Medicine, Department of Physiological Chemistry, Baltimore, Maryland 21205
Olusoji Amire, Department of Chemistry, University of Ibadan, Ibadan, Nigeria S. G . Angelos, Jr., Department of Chemistry, University of Southern Califor
nia, University Park, Los Angeles, California 90007 E. Antonini, Istituto di Chimica, Facolta di Medicina, Universita di Roma e
Centro di Biologia Molecolare, Rome, Italy Hiroyuki Arata, Department of Biology, Kyushu University, Fukuoka 812,
Japan Toshio Asakura, Department of Pediatrics, Children's Hospital of Philadelphia,
Philadelphia, Pennsylvania 19104 Winfried Ausländer, Max-Volmer Institut, Technische Universität Berlin,
Strasse des 17 juni 135, 1 Berlin 12, Federal Republic of Germany George Austin, Section of Neurosurgery, Loma Linda University School of
Medicine, Loma Linda, California 92354 Angelo Azz i , Universitität Bern, Med-Chem. Institut, Bühlstrasse 28, 3012
Bern, Switzerland G. F. Azzone, Instituto di Patologia Generale, Via Loredan, Padova, Italy Gerald T. Babcock, Department of Chemistry, Michigan State University, East
Lansing, Michigan
x v i i
AT/7/ Conthbutors o f Volumes I and II
B. Balko, Laboratory of Technical Development, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20014
Michael Barany, Department of Biological Chemistry and Research Resources Center, University of Illinois Medical Center, Chicago, Illinois 60612
Donald Barber, School of Biological Sciences, University of East Anglia, Nor-wich NR4 7TJ, England
J. Barber, Imperial College, Department of Botany, London, United Kingdom Clyde H . Barlow, Department of Biochemistry and Biophysics, Johnson Re
search Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
C. Lindsay Bashford, Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
B. Bean, Institute of Cancer Research, Fox Chase, Philadelphia, Pennsylvania 19111
John G . Beetlestone, Department of Chemistry, University of Ibadan, Ibadan, Nigeria
Helmut Beinert, Enzyme Institute, University of Wisconsin, Madison, Wisconsin 53706
Randy L . Bell , Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403
Nils-Olof Bengtsson, Department of Chemistry, Section of Physiological Chemistry, University of Umea, S-901 87 Umea, Sweden
Robert L . Berger, Building 10, Room 5D-20, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20014
S. Biddlecome, Division of Neurosurgery, Albany Medical College, Albany, New York
R. Bisson,.Istituto di Patologia Generale, Universita di Padova, Padova, Italy Bruno Blanchy, Universite Claude Bernard-Lyon 1, 69621 Villeurbanne,
France R. E. Blankenship, Department of Biochemistry, J405 Health Sciences Build
ing, SJ-70, University of Washington, Seattle, Washington 98195 Francis Blasco, Department of Human Genetics/Physiology, Yale University,
New Häven, Connecticut 06510 Kent Blasie, Departments of Chemistry and Biochemistry/Biophysics, Univer
sity of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104
Haywood Blum, Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, Medical School, Philadelphia, Pennsylvania 19104
Contributors of Volumes I and II x i x
W . E. Blumberg, Bell Telephone Laboratories, Murray Hi l l , New Jersey 07974 N . K . Boardman, CSIRO, P.O. Box 225, Dickson, A . C . T . 2602, Australia David F. Bocian, California Institute of Technology, Division of Chemistry and
Chemical Engineering, Pasadena, California 91125 Mireille Bof, Laboratoire de Biochimie, Departement de Recherche Fondamen
tale, C E N - 85X, 38041 Grenoble, Cedex, France D. Boffoli, Instituto di Chimica Biologica, Facolta di Medicina e Chirurgia,
Universita di Bari, 70126 Bari, Italy Arnold Boiteux, Max-Planck Institut für Ernährungsphysiologie, D-46
Dortmund, Rheinlanddamm 201, West Germany M . Bond, Department of Biochemistry and Biophysics, Johnson Research
Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Walter D. Bonner, Jr., Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Salil Bose, Department of Biology, University of Rochester, Rochester, New York 14627
R. S. Bourke, Division of Neurosurgery, Albany Medical College, Albany, New York
A . Boveris, Instituto de Quimica Biologica, Universidad de Buenos Aires, Buenos Aires, Argentina
P. Bowen, Laboratory of Technical Development, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20014
Barry J. Bowman, Department of Human Genetics/Physiology, Yale University, New Häven, Connecticut 06510
Charles Bowman, Department of Biological Sciences, State University of New York at Albany, Albany, New York 12222
M . K. Bowman, Chemistry Division D-200, Argonne National Laboratories, Argonne, Illinois 60439
John R. Bowyer, Department of Physiology and Biophysics, University of Illinois, Urbana, Illinois 61801
Paul D. Boyer, Department of Chemistry and Molecular Biology Institute, University of California at Los Angeles, Los Angeles, California 90024
Arthur S. Bri l l , Department of Physics, University of Virginia, Charlottesville, Virginia 22901
F. J. Brinley, Jr., Department of Physiology, University of Maryland, School of Medicine, Baltimore, Maryland 21201
A. M . Brown, Department of Physiology and Biophysics, University of Texas Medical Branch, Galveston, Texas 77550
XX Contributors of Volumes I and II
George Brown, Stanford Synchrotron Radiation Laboratory, Stanford, California
Truman R. Brown, Bell Laboratories, Murray Hi l l , New Jersey 07974 Mark Bruckner, Department of Biochemistry and Biophysics, Johnson Research
Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Gary W. Brudvig, California Institute of Technology, Division of Chemistry and Chemical Engineering, Pasadena, California 91125
M . Brunori, Institute of Biochemistry, Faculty of Medicine, Citta Universitaria, 00185 Rome, Italy
C. Tyler Burt, Department of Biological Chemistry and Research Resources Center, University of Illinois Medical Center, Chicago, Illinois 60612
Gerhard Buse, R W T H Aachen, Abt. Physiologische Chemie, Meiatener Strasse 211, D-5100 Aachen, West Germany
Roderick A. Capaldi, Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403
F. Capuano, Instituto di Chimica Biologica, Facolta di Medicina e Chirurgia, Universita di Bari, 70126 Bari, Italy
R. P. Casey, Universiteit van Amsterdam, Lab. voor Biochemie, B . C . P . Jarnsen Institut, Plantage Muidergracht 12, Amsterdam-C, Netherlands
Nicholas Chacos, Department of Biochemistry, Southwestern Medical School, University of Texas, Dallas, Texas 75235
Joseph M . Chalovich, Department of Biological Chemistry and Research Resources Center, University of Illinois Medical Center, Chicago, Illinois 60612
Samuel H . P. Chan, Department of Biology, Syracuse University, Syracuse, New York 13210
Sunney I. Chan, California Institute of Technology, Division of Chemistry and Chemical Engineering, Pasadena, California 91125
Britton Chance, Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
P. B . Chock, Laboratory of Biochemistry, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20014
W. S. Chow, CSIRO, Division of Plant Industry, Canberra, A . C . T . , Australia Leland C. Clark, Jr., Children's Hospital Research Foundation, Cincinnati,
Ohio 45229 Carol Coan, Laboratory of Physiology and Biophysics, University of the
Pacific, San Francisco, California 94115
Contributors o f Volumes I and II
Fredde Cohen, Department of Physiology and Biophysics, Health Sciences Center, State University of New York at Stonybrook, Stonybrook, New York 11794
L . B . Cohen, Department of Physiology, Yale University, School of Medicine, New Häven, Connecticut 06510
S. M . Cohen, Bell Laboratories, Murray Hi l l , New Jersey 07974 A . Colosimo, Istituto di Chimica, Facolta di Medicina, Universita di Roma e
Centro di Biologia Molecolare, Rome, Italy Pierre R. Coulet, Universite Claude Bernard-Lyon 1, 69621 Villeurbanne,
France John C. Cox, Department of Biochemistry, Medical Sciences Institute, Univer
sity of Dundee, Dundee DD1 4 H N , Scotland Anthony R. Crofts, Department of Physiology and Biophysics, University of
Illinois, Urbana, Illinois 61801 Michael A . Cusanovich, Department of Chemistry, University of Arizona, Tuc-
son, Arizona 85721 Morris J. Danon, Department of Biological Chemistry and Research Resources
Center, University of Illinois Medical Center, Chicago, Illinois 60612 A . Darszon, Departments of Physics and Biology, University of California at
San Diego, La Jolla, California 92037 P. H . Davis, Department of Physics and Center for Biological Macromolecules,
State University of New York at Albany, Albany, New York 12222 D. W. Deamer, Department of Zoology, University of California, Davis,
California 95616 Gilbert J. DeLeeuw, Department of Biochemistry and Biophysics, Johnson Re
search Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Leo De Maeyer, Max-Planck Institute für Biophysikalische Chemie, D-3400 Göttingen-Nikolausberg, West Germany
J. A . den Hollander, Bell Laboratories, Murray Hi l l , New Jersey 07974 Anne-Christine Dianous, Laboratoire de Biochimie, Departement de Recherche
Fondamentale, C E N - 8 5 X , 38041 Grenoble, Cedex, France M . Helena Mendonqa Dias, Centro de Quimica Estrutural, Universidade de
Lisboa, Instituto Superior Teenico, Lisbon, Portugal Richard E. Dickerson, Chemistry Department, California Institute of Technol
ogy, Pasadena, California 91125 James Dilger, Department of Physiology and Biophysics, Health Sciences
Center, State University of New York at Stonybrook, Stonybrook, New York 11794
.Y.Y/7 Contributors o f V o l u m es I and II
Richard A . Dilley, Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907
F. DiVirgilio, Instituto di Patologia Generale, Via Loredan, Padova, Italy Ed Dolan, Charles F. Kettering Research Laboratory, Yellow Springs, Ohio
45387 W. R. Dunham, Institute of Science and Technology, Biophysics Research Di
vision, University of Michigan, Ann Arbor, Michigan 48109 P. Leslie Dutton, Department of Biochemistry and Biophysics, Johnson Re
search Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
W. A . Eaton, Laboratory of Chemical Physics, National Institute of Arthritis, Metabolism, and Digestive Diseases, National Institutes of Health, Bethesda, Maryland 20014
J. F. Eccleston, Department of Biochemistry and Biophysics, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Moises Eisenberg, Department of Physiology and Biophysics, Health Sciences Center, State University of New York at Stonybrook, Stonybrook, New York 11794
Peter Eisenberger, Bell Laboratories, Murray Hi l l , New Jersey 07974 George Eisenman, Department of Physiology, School of Medicine, Center for
the Health Sciences, Los Angeles, California 90024 Laura Eisenstein, Department of Physics, University of Illinois, Urbana, Illinois
61801 S. Walter Englander, Department of Biochemistry and Biophysics, University of
Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104 Lars Ernster, Department of Biochemistry, Arrhenius Laboratory, University of
Stockholm, Stockholm, Sweden Ronald W. Estabrook, Department of Biochemistry, Southwestern Medical
School, University of Texas, Dallas, Texas 75235 M . C. W. Evans, Department of Botany and Microbiology, University College,
London W C IE 6BT, England James A . Fee, University of Michigan, Institute of Science and Technology,
Biophysics Research Division, Ann Arbor, Michigan 48109 Hubert Felle, Department of Physiology, Yale University School of Medicine,
New Häven, Connecticut 06510 F. A . Ferrone, Laboratory of Chemical Physics, N I A M D D , National Institutes
of Health, Bethesda, Maryland 20014 Francis G . Fiamingo, Department of Physics, University of Virginia, Charlot-
tesville, Virginia 22901
Contributors of Volumes I and II x x i i i
M . Filipkowski, Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
W . F. Filter, Institute of Science and Technology, Biophysics Research Division, University of Michigan, Ann Arbor, Michigan 48109
Karen L . Findling, Institute of Science and Technology, Biophysics Research Division, University of Michigan, Ann Arbor, Michigan 48109
S. Fleischer, Department of Molecular Biology, Vanderbilt University, Nashville, Tennessee 37235
Robert E . Forster, Department of Physiology, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Hans Frauenfelder, Department of Physics, University of Illinois, Urbana, Illinois 61801
Jeffrey C. Freedman, Department of Physiology, Yale University School of Medicine, New Häven, Connecticut 06510
Terrence G . Frey, Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Luciano Frigeri, Scripps Clinic and Research Foundation, Department of Biochemistry, La Jolla, California 92037
David G . Gadian, Department of Biochemistry, University of Oxford, Oxford O X l 3QU, England
Yves M . Galante, Scripps Clinic and Research Foundation, La Jolla, California 92037
Peter B . Garland, Department of Biochemistry, Medical Sciences Institute, University of Dundee, Dundee DD1 4 H N , Scotland
Pamela B . Garlick, Department of Biochemistry, University of Oxford, Oxford, O X l 3QU, England
Daniele C. Gautheron, University of Claude Bernard-Lyon 1, 69621 Villeur-banne, France
Elzbieta Glaser, Department of Biochemistry, Arrhenius Laboratory, University of Stockholm, Stockholm, Sweden
Peter Glatz, Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Thomas Glonek, Department of Biological Chemistry and Research Resources Center, University of Illinois Medical Center, Chicago, Illinois 60612
Catherine Godinot, Universite Claude Bernard-Lyon 1, 69621 Villeurbanne, France
xxiv Contributors o f Volumes I and II
Sidney L . Gordon, Institut für Molekularbiologie und Biophysik, E T H -Zürich-Honggerberg, CH-8093 Zürich, Switzerland
B. I. Greene, Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania 19104
J. A . Greenrod, Department of Biochemistry, University of Birmingham, Birmingham B15 2TT, United Kingdom
Colin Greenwood, University of East Anglia, School of Biological Sciences, Norwich N R 4 7TJ, England
Ferdinand Greitschus, Max Planck Institut für Systemphysiologie, D-4600 Dortmund, West Germany
A . Grinvald, Department of Physiology, Yale University, School of Medicine, New Häven, Connecticut 06510
A . K . Groen, Universiteit van Amsterdam, Lab. voor Biochemie, B . C . P . Jarnsen Institut, Plantage Muidergracht 12, Amsterdam-C, Netherlands
Markus Güggi, Max-Planck Institut für Systemphysiologie, D-4600 Dortmund, West Germany
F. Guerrieri, Instituto di Chimica Biologica, Facolta di Medicina e Chirurgia, Universita di Bari, 70126 Bari, Italy
H . Gutweniger, Istituto di Patologia Generale, Universita di Padova, Padova, Italy
David D . Hackney, Department of Chemistry and Molecular Biology Institute, University of California at Los Angeles, Los Angeles, California 90024
Bunji Hagihara, Department of Biochemistry, Osaka University Medical School, Osaka, Japan
David O. Hall, Department of Plant Sciences, King's College, London, England
Don A . Hampton, Alabama Power Company, Birmingham, Alabama 35291 Wesley Harden III, Harrison Department of Surgical Research, University of
Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104 L . J. Harding, Computing Center, University of Michigan, Ann Arbor, Michi
gan 48109 Alden H . Harken, Harrison Department of Surgical Research, University of
Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104 John Haselgrove, Department of Biochemistry and Biophysics, Johnson Re
search Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Youssef Hatefi, Department of Biochemistry, Scripps Clinic and Research Foundation, La Jolla, California 92037
Norio Hayashi, First Department of Internal Medicine, Osaka University Medical School, Osaka, Japan
Contributors of Volumes I and II xxv
P. Heathcote, Department of Botany and Microbiology, University College, London W C IE 6BT, England
Michele F. Henry, C N R S , DRF/Biochemie C E N - G 85X 38041 Grenoble, Cedex, France
Deborah K . Henson, Department of Chemistry, Indiana Universi ty, Bloomington, Indiana 47401
L . Herbette, Departments of Chemistry & Biochemistry/Biophysics, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Benno Hess, Max-Planck Institut für Ernährungsphysiologie, Rheinlanddamm 201, D-4600 Dortmund 1, West Germany
Hajime Hirata, Jichi Medical School, Tochigi-ken, Japan 329-04 George Hoch, Department of Biology, University of Rochester, Rochester,
New York 14627 R. M . Hochstrasser, Department of Chemistry, University of Pennsylvania,
Philadelphia, Pennsylvania 19104 Joseph F. Hoffman, Department of Physiology, Yale University School of
Medicine, New Häven, Connecticut 06510
J. Hofrichter, Laboratory of Chemical Physics, National Institute of Arthritis, Metabolism, and Digestive Diseases, National Institutes of Health, Bethesda, Maryland 20014
D. Holten, Department of Chemistry, Washington State University, Pullman, Washington
Jens Höper, Max-Planck Institut für Systemphysiologie, D-4600 Dortmund, West Germany
H . P. Hopkins, Department of Chemistry, Georgia State University, Atlanta, Georgia
Cheng-Schen Huang, Department of Biochemistry, School of Medicine, Wayne State University, Detroit, Michigan 48201
R. Lee Hutton, Department of Chemistry and Molecular Biology Institute, University of California at Los Angeles, Los Angeles, California 90024
James S. Hyde, Department of Biochemistry, Medical College of Wisconsin, Milwaukee County Medical Complex, Milwaukee, Wisconsin 53226
Vito Iacobazzi, Department of Biochemistry, University of Bari, Via Amendola 165, Bari, Italy
Masao Ideda-Saito, Department of Biochemistry and Biophysics, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Giuseppe Inesi, Laboratory of Physiology and Biophysics, University of the Pacific, San Francisco, California 94115
x.xv i Contributors of Volumes I and II
W. John Ingledew, Department of Biochemistry, Medical Sciences Institute, University of Dundee, Dundee DD1 4 H N , Scotland
Toshiro Inubushi, Department of Biochemistry and Biophysics, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Nobutomo Itada, Department of Physiology, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
G. Izzo, Instituto di Chimica Biologica, Facolta di Medicina e Chirurgia, Universita di Bari, 70126 Bari, Italy
J. B. Jackson, Department of Biochemistry, University of Binningham, Birmingham B15 2TT, United Kingdom
Arco Y . Jeng, Department of Radiation Biology and Biophysics, University of Rochester, Rochester, New York 14642
Frans F. Jöbsis, Department of Physiology and Pharmacology, Duke University Medical Center, Durham, North Carolina 27710
Michael Johnson, School of Chemical Sciences, University of East Anglia, Norwich NR4 7TJ, England
Robert G. Johnson, Department of Biochemistry and Biophysics, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Robert W. Jones, Department of Biochemistry, Medical Sciences Institute, University of Dundee, Dundee DD1 4 H N , Scotland
Elizabeth Joyce, Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Wolfgang Junge, Max-Volmer Institut, Technische Universität Berlin, Strasse des 17 juni 135, 1 Berlin 12, Federal Republic of Germany
Ronald Jutzy, Section of Neurosurgery, Loma Linda University School of Medicine, Loma Linda, California 92354
H . R. Kaback, Roche Institute of Molecular Biology, Nutley, New Jersey 07110
Gregory J. Kaczorowski, Laboratory of Membrane Biochemistry, Roche Institute of Molecular Biology, Nutley, New Jersey 07110
Yasuo Kagawa, Jichi Medical School, Tochigi-ken, Japan 329-04 Takenobu Kamada, First Department of Internal Medicine, Osaka University
Medical School, Osaka, Japan Hilde Kanamuller, Department of Biochemistry and Biophysics, Johnson Re
search Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Sunao Kawano, Department of Biochemistry, Southwestern Medical School, University of Texas, Dallas, Texas 75235
Contributors o f Volumes I and II .x.xvii
Bacon Ke, Charles F. Kettering Research Laboratory, Yellow Springs, Ohio 45387
Regula M . Keller, Institut für Molekularbiologie und Biophysik, E T H -Zürich-Hönggerberg, CH-8093 Zürich, Switzerland
Manfred Kessler, Max-Planck-Insti tüt für Systemphysiologie, D-4600 Dortmund, West Germany
Ezzatollah Keyhani, Laboratory for Cell Biology and Biochemistry, Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran
Jacqueline Keyhani, Laboratory for Cell Biology and Biochemistry, Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran
H . K . Kimelberg, Division of Neurosurgery, Albany Medical College, Albany, New York 12222
B. M . Kincaid, Bell Laboratories, Murray Hi l l , New Jersey 07974 Tsoo E. King, Department of Chemistry and Laboratory of Bioenergetics, State
University of New York at Albany, Albany, New York 12222 Gerard Klein, Laboratoire de Biochimie, Departement de Recherche Fondamen
tale, C E N 85X, 38041 Grenoble, Cedex, France Amira Klip, C . H . Best Institute, University of Toronto, Toronto, Ontario,
Canada M 5 G 1L6 Josephine L . Y . Kong, Department of Chemistry, Northwestern University,
Evanston, Illinois 60301 W. H . Koppenol, Department of Biochemistry and Molecular Biology, North
western University, Evanston, Illinois 60201 Harold Korb, Physiologisch-Chemisches Institut der Georg-August-Universität
Göttingen, 34 Göttingen, Humboldtallee 7, West Germany R. Koren, Institute for Cancer Research, Fox Chase, Philadelphia, Pennsyl
vania 19111 Arthur Kowalsky, Department of Biophysics, Albert Einstein College of
Medicine, Yeshiva University, Bronx, New York 10461 Klaas Krab, Department of Medical Chemistry, University of Helsinki, Sil-
tavuorenpenger 10, SF 00170 Helsinki 17, Finland Mark Kurzmack, Laboratory of Physiology and Biophysics, University of the
Pacific, San Francisco, California 94115 Dietrich Kuschmitz, Max-Planck Institut für Ernährungsphysiologie, Rhein
landdamm 201, D-4600 Dortmund 1, West Germany Henry Lardy, Institute for Enzyme Research, University of Wisconsin, Madi-
son, Wisconsin 53706 Frank A . Lattanzio, Department of Pharmacology, University of Miami School
of Medicine, P.O. Box 520875, Miami, Florida 33152
.v.vv/77 Contributors of Volumes I and II
Pui-Wah Lau, Department of Biochemistry and Biophysics, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Paul C. Lauterbur, Department of Chemistry, State University of New York at Stonybrook, Stonybrook, New York 11794
C. P. Lee, Biochemistry Department, Wayne State University, School of Medicine, Detroit, Michigan 48201
K. S. Lee, Department of Physiology and Biophysics, University of Texas Medical Branch, Galveston, Texas 77550
Alison Lees, Institute of Science & Technology, Biophysics Research Division, University of Michigan, Ann Arbor, Michigan 48109
Albert L . Lehninger, Department of Physiological Chemistry, Johns Hopkins University, School of Medicine, Baltimore, Maryland 21205
J. S. Leigh, Jr., Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Aaron Lewis, Clark Hall, Cornell University, School of Applied and Engineering Physics, Ithaca, New York 14853
David E. Lewis, Laboratory of Physiology and Biophysics, University of the Pacific, San Francisco, California 94115
R. A . Lieberman, Biophysics Research Division, Institute of Science and Technology, University of Michigan, Ann Arbor, Michigan 48109
David Y . Lo , Department of Genetics, Stanford University, School of Medicine, Stanford, California 94305
Paul A . Loach, Department of Biochemistry and Molecular Biology, Northwestern University, Evanston, Illinois 60301
Gilda Loew, Department of Genetics, Stanford University, School of Medicine, Stanford, California 94305
W. Scott Long, Department of Physiology, Yale University School of Medicine, New Häven, Connecticut 06510
M . Lorusso, Instituto di Chimica Biologica, Facolta di Medicina e Chirurgia, Universita di Bari, 70126 Bari, Italy
Bernd Ludwig, Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403
Martha L . Ludwig, Biophysics Research Division and Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48109
Joel Lunardi, Laboratoire de Biochimie, Departement de Recherche Fondamentale, C E N - 8 5 X , 38041 Grenoble, Cedex, France
Elizabeth McCandlish, Department of Chemistry, Rutgers University, Busch Campus, New Brunswick, New Jersey 08903
Contributors of Volumes I and II xv/.v
Richard E. McCarty, Section of Biochemistry, Molecular, and Cell Biology, Cornell University, Ithaca, New York 14853
George G . McDonald, Department of Biochemistry and Biophysics, University of Pennsylvania Medical School, Johnson Research Foundation, Philadelphia, Pennsylvania 19104
Allison McGeer, Max-Volmer Institut, Technische Universität Berlin, Strasse des 17 juni 135. 1 Berlin 12, Federal Republic of Germany
A . C . McLaughlin, Biology Department, Brookhaven National Laboratory, Up-ton, New York 11973
Stuart McLaughlin, Department of Physiology and Biophysics, Health Sciences Center, State University of New York at Stonybrook, Stonybrook, New York 11794
David H . MacLennan, C. H . Best Institute, University of Toronto, Toronto, Ontario, Canada M5G 1L6
Ronald P. Magnusson, Section of Biochemistry, Molecular, and Cell Biology, Cornell University, Ithaca, New York 14853
Henry R. Mahler, Department of Chemistry, Indiana University, Bloomington, Indiana 47401
T. Maida, Department of Biochemistry and Biophysics, University of Pennsylvania, School of Medicine, Johnson Research Foundation, Philadelphia, Pennsylvania 19104
Bruce L . Maloff, Department of Biological Sciences, State University of New York at Albany, Albany, New York 12222
Rimona Margalit, Department of Physiology, School of Medicine, Center for the Health Sciences, Los Angeles, California 90024
E. Margoliash, Department of Biochemistry and Molecular Biology, Northwestern University, Evanston, Illinois 60201
Takakatsu Matsumura, First Department of Internal Medicine, Osaka University Medical School, Osaka, Japan
Ichiro Matsuoka, Department of Biology, Faculty of Science, Osaka University, Toyonaka, Osaka 560, Japan
Avraham Mayevsky, Department of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel
J. Maylie, Department of Physiology, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Werner Meesman, Pathologisches Institut, Universität Essen, Essen, West Germany
V. Miconi, Instituto di Patologia Generale, Via Loredan, Padova, Italy E. V . Mielczarek, Laboratory of Technical Development, National Heart,
A'.V.V Contributors o f Volu/nes I and II
Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20014
A . S. Mildvan, Institute of Cancer Research, Fox Chase, Philadelphia, Pennsylvania 19111
Donald H . Miller, Department of Chemistry, Indiana University, Bloomington, Indiana 47401
Peter Mitchell, Glynn Research Laboratories, Bodmin, Cornwall PL30 4 A U , England
Roy Mitchell, Glynn Research Laboratories, Bodmin, Cornwall PL30 4 A U , England
M . Montal, Physics Department, University of California, San Diego, La Jolla, California 92037
C. Montecucco, Istituto di Patologia Generale, Universita di Padova, Padova, Italy
Anthony L . Moore, Department of Plant Sciences, King's College, London, England
M . Morad. Department of Physiology, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Randall H . Morse, California Institute of Technology, Division of Chemistry and Chemical Engineering, Pasadena, California 91125
Jennifer Moyle, Glynn Research Laboratories, Bodmin, Cornwall PL30 4 A U , England
S. Nagaoka, Department of Biochemistry, State University of New York at A l bany, Albany, New York 12222
Takao Nakamura, Department of Biology, Faculty of Science, Osaka University, Toyonaka, Osaka 560, Japan
Y . Nakase, Department of Biochemistry and Biophysics, University of Pennsylvania, Johnson Foundation, School of Medicine, Philadelphia, Pennsylvania 19104
Walter Neupert, Physiologisch-Chemisches Institut der Georg-August-Universität Göttingen, 34 Göttingen, Humboldtallee 7, West Germany
David Nicholls, University of Dundee, Department of Psychiatry, Ninewells Hospital, Dundee DD1 9SY, Scotland
Peter Nicholls, Department of Biological Sciences, Brock University, Glenridge Campus, St. Catharines, Ontario L2S 3A1, Canada
J. Wylie Nichols, Department of Zoology, University of California, Davis, California 95616
Mitsuo Nishimura, Department of Biology, Kyushu University, Fukuoka, 812 Japan
Contributors of Volumes I and II
Birgitta Norling, Department of Biochemistry, Arrhenius Laboratory, University of Stockholm, Stockholm, Sweden
J. R. Norris, Chemistry Division D-200, Argonne National Laboratories, Argonne, Illinois 60439
Barry E. North, Biology Department, Brookhaven National Laboratory, Upton, New York 11973
Svante Nyberg, Department of Chemistry, Section of Physiological Chemistry, University of Umea, S-901 87 Umea, Sweden
S. Ogawa, Bell Laboratories, Murray Hi l l , New Jersey 07974 Gabriel B . Ogunmola, Department of Chemistry, University of Ibadan, Ibadan,
Nigeria Per-Ingvar Ohlsson, Department of Chemistry, Section of Physiological
Chemistry, University of Umea, S-901 87 Umea, Sweden Tomoko Ohnishi, Department of Biochemistry and Biophysics, Johnson Re
search Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Yutaka Orii , Department of Biology, Faculty of Science, Osaka University, Toyonaka, Osaka, Japan
Donald R. Ort, Department of Biochemistry, University of Washington, Seattle, Washington 98195
Neil Osheroff, Department of Biochemistry and Molecular Biology, Northwestern University, Evanston, Illinois 60201
Charles S. Owen, Department of Biochemistry and Biophysics, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
James Pachence, Departments of Chemistry and Biochemistry/Biophysics, University of Pennsylvania Medical School, Philadelphia, Pennsylvania 19104
N . K. Packham, Department of Biochemistry, University of Birmingham, Birmingham B15 2TT, United Kingdom
Ferdinando Palmieri, Department of Biochemistry, University of Bari, Via Amendola 165, Bari, Italy
Angela Pansini, Instituto di Chimica Biologica, Facolta di Medicina e Chirur-gia, Universita di Bari, 70126 Bari, Italy
S. Papa, Instituto di Chimica Biologica, Facolta di Medicina e Chirurgia, Universita di Bari, 70126 Bari, Italy
William W. Parson, Department of Biochemistry, J405 Health Sciences Building, SJ-70, University of Washington, Seattle, Washington 98195
Donald F. Parsons, Electron Optics Laboratory, State of New York Department of Health, Tower Building, Empire State Plaza, Albany, New York 12201
xxx ii Contributors o f Volumes / and II
Lehka Patel, Laboratory of Membrane Biochemistry, Roche Institute of Molecular Biology, Nutley, New Jersey 07110
Katherine A . Pattridge, Biophysics Research Division and Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48109
Karl Gustav Paul, Department of Medical Chemistry, University of Umea, S-901 87 Umea, Sweden
R. Paul, Laboratory of Technical Development, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20014
Freddy Paulussen, Max-Planck Institut für Biophysikalische Chemie, D-3400 Göttingen-Nikolausberg, West Germany
Laraine Peiffer, Department of Physiology, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Jack Peisach, Department of Molecular Pharmacology, Albert Einstein College of Medicine, Yeshiva University, Bronx, New York 10461
R. A . Perreault, Department of Chemistry, University of Southern California, University Park, Los Angeles, California 90007
Gregory A . Petsko, Department of Biochemistry, Wayne State University School of Medicine, Detroit, Michigan 48201
K . M . Petty, Department of Biochemistry, University of Birmingham, Birmingham B15 2TT, United Kingdom
R. M . Polichar, Science Applications, Inc., La Jolla, California Carol A . Popp, National Biomedical ESR Center, Department of Radiology,
Medical College of Wisconsin, Milwaukee, Wisconsin 53226 Richard Pougeois, Laboratoire de Biochimie, Departement de Recherche Fon
damentale, C E N - 85X, 38041 Grenoble, Cedex, France Linda Powers, Bell Telephone Laboratories, Murray H i l l , New Jersey 07974 T. Pozzan, Instituto di Patologia Generale, Via Loredan, Padova, Italy Berton C. Pressman, Department of Pharmacology, University of Miami School
of Medicine, P.O. Box 520875, Miami, Florida 33152 Roger C. Prince, Department of Biochemistry and Biophysics, Johnson Re
search Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
L . J. Prochaska, Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907
Alexandre T. Quintanilha, Membrane Bioenergetics Group, Lawrence Berkeley Laboratory, University of California, Berkeley, California 94720
Bjgrrn Quistorff, Department of Biochemistry A , Panum Institute, University of Copenhagen, DK-2200 Copenhagen N , Denmark
Contributors o f Volumes I and II x x x i i i
Edd Rabon, Laboratory of Membrane Biology, University of Alabama at Birmingham, Birmingham, Alabama 35294
Efraim Racker, Section of Biochemistry, Molecular, and Cell Biology, Cornell University, Ithaca, New York 14853
George K . Radda, Department of Biochemistry, University of Oxford, Oxford O X l 3 Q U , England
Hassan Rastegar, Harrison Department of Surgical Research, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
P. M . Rentzepis, Bell Laboratories, Murray Hi l l , New Jersey 07974 Baltazar Reynafarje, Department of Physiological Chemistry, Johns Hopkins
University, School of Medicine, Baltimore, Maryland 21205 S. G . Rhee, Laboratory of Biochemistry, National Heart, Lung, and Blood In
stitute, National Institutes of Health, Bethesda, Maryland 20014 Peter R. Rieh, Department of Biochemistry, University of Cambridge, Cam
bridge, England Lajos Rimai, Research Staff, Ford Motor Company, Dearborn, Michigan
48121 Dan E . Robertson, Laboratory of Membrane Biochemistry, Roche Institute of
Molecular Biology, Nutley, New Jersey 07110 David Rose, Department of Biochemistry, Wayne State University, School of
Medicine, Detroit, Michigan 48201 W. N . Ross, Department of Neurobiology, Harvard University, Boston, Mas
sachusetts Hagai Rottenberg, Bell Laboratories, Murray Hi l l , New Jersey 07974 Andrew M . Rudin, Northport Veterans Administration Hospital, Northport,
New York Heinz Rupp, Department of Plant Sciences, King's College, London, England Frank J. Ruzicka, Enzyme Institute, University of Wisconsin, Madison, Wis
consin 53706 Gaetano Saccomani, Laboratory of Membrane Biology, University of Alabama
at Birmingham, Birmingham, Alabama 35294 Roland Sacher, R W T H Aachen, Abt. Physiologische Chemie, Melatener
Strasse 211, D-5100 Aachen, West Germany George Sachs, Laboratory of Membrane Biology, University of Alabama at
Birmingham, Birmingham, Alabama 35294 A. Saito, Department of Molecular Biology, Vanderbilt University, Nashville,
Tennessee 37235 F. R. Salemme, Department of Chemistry, University of Arizona, Tucson,
Arizona 85721
xxxiv Contributors o f Volumes I and II
John C. Salerno, Department of Biochemistry, Duke University, School of Medicine, Durham, North Carolina
L . Salganicoff, Department of Pharmacology and Specialized Center for Thrombosis Research, Temple University, Philadelphia, Pennsylvania
Irving Salmeen, Ford Motor Company, Dearborn, Michigan 48121 B. M . Salzberg, Department of Physiology and Pharmacology, School of Dental
Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104 R. H . Sands, University of Michigan, Institute of Science and Technology,
Biophysics Research Division, Ann Arbor, Michigan N - E Leo Saris, Department of Medical Chemistry, University of Helsinki, Si l-
tavuorenpenger 10, SF 00170 Helsinki 17, Finland Nobuhiro Sato, First Department of Internal Medicine, Osaka University Medi
cal School, Osaka, Japan Michael Satre, Laboratoire de Biochimie, Departement de Recherche Fon
damentale, Dentre d'Etudes Nucleaires, 85X, 38041 Grenoble Cedex, France
Antonio Scarpa, Department of Biochemistry and Biophysics, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Günter Schäfer, Med. Hochschule Hanover, Postfach 610 180, 3 Hanover 61, West Germany
Gottfried Schatz, Biocenter, University of Basel, CH-4056 Basel, Switzerland W. Robert Scheidt, Department of Chemistry, University of Notre Dame, Notre
Dame, Indiana 46556 C. C. Schenck, Department of Biochemistry, J405 Health Sciences Building,
SJ-70, University of Washington, Seattle, Washington 98195 Benno P. Schoenborn, Brookhaven National Laboratory, Upton, New York
11973 Charles P. Scholes, Department of Physics, State University of New York at
Albany, Albany, New York 12222 S. Schreier, Department of Bioquimica, Instituto de Quimica, Universidade de
Säo Paulo, Säo Paulo, Brazil Dana M . Scott, Department of Physiology, University of Pennsylvania, School
of Medicine, Philadelphia, Pennsylvania 19104 P. John Seeley, Department of Biochemistry, University of Oxford, Oxford
O X l 3QU, England C. H . A . Seiter, Department of Chemistry, University of Southern California,
University Park, Los Angeles, California 90007 Adil E. Shamoo, Department of Radiation Biology and Biophysics, University
of Rochester, Rochester, New York 14642
Contributors o f Volutnes I and II xxxv
C. V . Shank, Bell Telephone Laboratories, Holmdel, New Jersey Motoaki Shichiri, First Department of Internal Medicine, Osaka University
Medical School, Osaka, Japan R. G . Shulman, Bell Laboratories, Murray, H i l l , New Jersey 07974 H . Shuman, Departments of Physiology and Pathology, University of Pennsyl
vania, School of Medicine, Philadelphia, Pennsylvania 19104 Vladimir. A . Shuvalov, Charles F. Kettering Research Laboratory, Yellow
Springs, Ohio 45387 Helmut Sies, Institut für Physiologische Chemie, Physikalische Biochemie, and
Zellbiologie, Universität München 2, Munich, West Germany Binah R. Silberstein, Department of Biochemistry and Biophysics, Johnson Re
search Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Ian A . Silver, Department of Pathology, University of Bristol Medical School, Bristol BS8 1TD, England
Michael B . Simson, Department of Cardiology, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Carolyn W. Slayman, Department of Human Genetics, Yale University, New Häven, Connecticut 06510
Clifford L . Slayman, Department of Physiology, Yale University School of Medicine, New Häven, Connecticut 06510
Jerry Smith, Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Ward W. Smith, Molecular Biology Institute, University of California at Los Angeles, Los Angeles, California
A. P. Somlyo, Pennsylvania Muscle Institute, Philadelphia, Pennsylvania 19104
A. V . Somlyo, Pennsylvania Muscle Institute, Philadelphia, Pennsylvania 19104
Nobuhito Sone, Jichi Medical School, Tochigi-ken, Japan 329-04 Annick Soumarmon, Section of Biochemistry, Molecular, and Cell Biology,
Cornell University, Ithaca, New York 14853 Thomas G . Spiro, Department of Chemistry, Princeton University, Princeton,
New Jersey 08540 John Springall, School of Chemical Sciences, University of East Anglia, Nor-
wich NR4 7TJ, England B. R. Sreenathan, Department of Biophysics, University of Western Ontario,
Health Sciences Center, London N6A 5C1, Canada
xxxvi Contributors of Volumes I and II
E. R. Stadtman, Laboratory of Biochemistry, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20014
James Stamatoff, Bell Telephone Laboratories, Murray H i l l , New Jersey 07974 Charles Steenbergen, Department of Biochemistry and Biophysics, Johnson Re
search Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Gerd J. Steffens, R W T H Aachen, Abt. Physiologische Chemie, Melatener Strasse 211, D-5100 Aachen, West Germany
Guy C. M . Steffens, R W T H Aachen, Abt. Physiologische Chemie, Melatener Strasse 211, D-5100 Aachen, West Germany
P. Stein, Institute for Cancer Research, Fox Chase, Philadelphia, Pennsylvania 19111
Paul Stein, Department of Chemistry, Princeton University, Princeton, New Jersey 08540
David L . Stetson, Department of Physiology, Yale University School of Medicine, New Häven, Connecticut 06510
Tom H . Stevens, California Institute of Technology, Division of Chemistry and Chemical Engineering, Pasadena, California 91125
Italo Stipani, Department of Biochemistry, University of Bari, Via Amendola 165, Bari, Italy
John D. Stong, Department of Chemistry, Princeton University, Princeton, New Jersey 08540
Bayard T. Storey, Department of Physiology, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
R. Strasser, Departments of Physics and Biology, University of California at San Diego, La Jolla, California 92037
Peter Styles, Department of Biochemistry, University of Oxford, Oxford O X l 3QU, England
Jeanne Sweetland, Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403
Ken-Ichiro Takamiya, Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Tsunehiro Takano, Department of Chemical Engineering, California Institute of Technology, Pasadena, California 91125
Hiroshi Takeda, Department of Surgery, Kyoto University, Medical School, Kyoto, Japan
C. P. S. Taylor, Department of Biophysics, University of Western Ontario, Health Sciences Center, London N6A 5C1, Canada
Contributors o f Volumes I and II xxxv i i
Henry Tedeschi, Department of Biological Sciences, State University of New York at Albany, Albany, New York 12222
A . Telfer, Department of Botany, Imperial College, London, United Kingdom
Andrew Thomson, School of Chemical Sciences, University of East Anglia, Norwich NR4 7TJ, England
S. W. Thorne, CSIRO, Division of Plant Industry, Canberra, A . C . T . , Aus-tralia
M . C. Thurnauer, Chemistry Division, D-200, Argonne National Laboratories, Argonne, Illinois 60439
D. M . Tiede, Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
J. T. Tiffert, Department of Physiology, University of Maryland, School of Medicine, Baltimore, Maryland 21201
William F. Tivol, Department of Radiation Biology and Biophysics, University of Rochester, Rochester, New York 14642
D. R. Trentham, Department of Biochemistry and Biophysics, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
A. D. Trifunac, Chemistry Division D-200, Argonne National Laboratories, Argonne, Illinois 60439
Bernard L . Trumpower, Dartmouth Medical School, Hanover, New Hampshire 03755
Demetrius Tsernoglou, Department of Biochemistry, Wayne State University, School of Medicine, Detroit, Michigan 48201
K. Ugurbil, Bell Laboratories, Murray Hi l l , New Jersey 07974 D. W. Urry, Laboratory of Molecular Biophysics, University of Alabama Med
ical Center, University Station, Birmingham, Alabama 35294 Joan S. Valentine, Department of Chemistry, Rutgers University, Busch Cam
pus, New Brunswick, New Jersey 08903 H. L . Van Camp, Biophysics Research Laboratory, University of Michigan,
Ann Arbor, Michigan 48104 K. van Dam, Universiteit van Amsterdam, Lab. voor Biochemie, B . C . P . Jarn
sen Institut, Plantage Muidergracht 12, Amsterdam-C, Netherlands Willem H . van den Berg, Department of Biochemistry and Biophysics, Johnson
Research Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Jane Vanderkooi, Department of Biochemistry and Biophysics, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
x x x v i i i Contributors o f Volumes I and II
R. van der Meer, Universiteit van Amsterdam, Lab. voor Biochemie, B . C . P . Jarnsen Institut, Plantage Muidergracht 12, Amsterdam-C, Netherlands
Sergio Verjovski-Almeida, Laboratory of Physiology and Biophysics, University of the Pacific, San Francisco, California 94115
Paulette M . Vignais, Maitre de Recherche au C N R S , DRF/Biochemie C E N - G 85X, 38041 Grenoble, Cedex, France
Pierre V. Vignais, Laboratoire de Biochimie, Departement de Recherche Fondamentale, C E N - 85X, 38041 Grenoble, Cedex, France
Steven Vik , Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403
John L . Walker, Department of Physiology, University of Utah College of Medicine, Salt Lake City, Utah 84108
Terrence Walsh, School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, England
C. T. Wang, Department of Molecular Biology, Vanderbilt University, Nashville, Tennessee 37235
A . Warshel, Department of Chemistry, University of Southern California, University Park, Los Angeles, California 90007
Michael R. Wasielewski, Chemistry Division, Argonne National Laboratory, Argonne, Illinois 60439
Takahide Watanabe, Department of Biology, Faculty of Science, Osaka University, Toyonaka, Osaka 560, Japan
Watt W. Webb, School of Applied and Engineering Physics, Cornell University, Ithaca, New York 14853
Gregorio Weber, Roger Adams Laboratory, University of Illinois, Urbana, Illinois 61801
Y . H . Wei, Laboratory of Bioenergetics, State University of New York at A l bany, Albany, New York 12222
R. B. Weisman, Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania 19104
J. Weiss, Department of Physiology, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
R. Weiss, Department of Chemistry, University of Southern California, University Park, Los Angeles, California 90007
Jürgen Werringloer, Department of Biochemistry, Southwestern Medical School, University of Texas, Dallas, Texas 75235
H . V . Westerhoff, Universiteit van Amsterdam, Lab. voor Biochemie, B . C . P . Jarnsen Institut, Plantage Muidergracht 12, Amsterdam-C, Netherlands
Contributors o f Volumes I and II x x x i x
W. R. Widger, Department of Chemistry and Laboratory of Bioenergetics, State University of New York at Albany, Albany, New York 12222
Volker Wiegand, Pathologisches Institut, Universität Essen, Essen, West Germany
Märten Wikström, Department of Medical Chemistry, University of Helsinki, Siltavuorenpenger 10, SF 00170 Helsinki 17, Finland
John R. Williamson, Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
M . T. Wilson, Department of Chemistry, University of Essex, Colchester, United Kingdom
M . W. Windsor, Department of Chemistry, Washington State University, Pullman, Washington
Raymond Wong, Section of Biochemistry, Molecular, and Cell Biology, Cornell University, Ithaca, New York 14853
Colin A . Wraight, Department of Botany, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801
Kurt Wüthrich, Institute für Molekularbiologie und Biophysik, E T H -Hönggerberg, CH-8093 Zürich, Switzerland
S. Yasui, Department of Physiology and Biophysics, University of Texas Medical Branch, Galveston, Texas 77550
Takashi Yonetani, Department of Biochemistry and Biophysics, Johnson Research Foundation, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania 19104
Masasuke Yoshida, Jichi Medical School, Tochigi-ken, Japan 329-04 Tatsuro Yoshida, Institute of Science and Technology, Biophysics Research Di
vision, University of Michigan, Ann Arbor, Michigan 48109 C. A . Y u , Department of Biochemistry, State University of New York at A l
bany, Albany, New York 12222 L. Y u , Department of Chemistry and Laboratory of Bioenergetics, State Univer
sity of New York at Albany, Albany, New York 12222 Richard Zimmermann, Physiologisch-Chemisches Institut der Georg-August-
Universität Göttingen, 34 Göttingen, Humboldtallee 7, West Germany
F R O N T I E R S O F BIOLOGICAL. E N E R G E T I C S , V O L U M E I
BIOSYNTHESIS AND INTRACELLULAR TRANSLOCATION OF MITOCHONDRIAL PROTEINS: CYTOCHROME C AND THE
CARBOXYATRACTYLOSIDE BINDING PROTEIN
Richard Zimmermann, Harald Korb and Walter Neupert
I n s t i t u t für Physiologische Chemie Universität Göttingen
Göttingen, GFR
I. INTRODUCTION
The vast majority of mitochondrial proteins i s t r a n s l a t e d on cytoplasmic ribosomes and must eventually be transported to t h e i r s i t e of function i n the mitochondrion. In previous studi e s we have presented evidence f or a tran s f e r mechanism which involves extramitochondrial precursors (Hallermayer et a l . , 1977; Harmey et a l . , 1977; Zimmermann et a l . , 1977) . These studies strongly argued against the proposal that as a general r u l e proteins are tra n s f e r r e d to mitochondria by d i r e c t Ins e r t i o n of nascent Polypeptide chains, c a r r i e d out by a sp e c i a l c l a s s of cytoplasmic. ribosomes bound to the outer mitochondrial membrane (Kellems and Butow, 1972; Kellems et a l . , 1 9 7 5 ) .
Attempts to further elucidate the mechanisms of tr a n s f e r have to take i n t o consideration that mitochondrial proteins d i f f e r widely with respect to t h e i r structure, physico-chemical properties and submitochondrial l o c a t i o n . Proteins of the outer membrane, the intermembrane Space, the inner membrane (peripheral and in t e g r a l ) and of the matrix space have to be distin g u i s h e d . I t may be expected that the mechanisms of tran s f e r are d i f f e r e n t f o r these various groups of protein s . Accordingly, the tra n s f e r processes have to be studied f o r i n -d i v i d u a l proteins of each group. The primary t r a n s l a t i o n products have to be analysed, extramitochondrial precursors must be i s o l a t e d , t h e i r s u b c e l l u l a r l o c a t i o n determined and th e i r s t r u c t u r a l r e l a t i o n s h i p to the fun c t i o n a l proteins
Copyright © 1978 by Academic Press, Inc. All rights of reproduction in any form reserved.
ISBN 0-12-225401-5
P a r t I E/cctrochcmical Intcractions 147
investigated. F i n a l l y , the mechanism of transfer of extramitochondrial precursors must be studied i n reconstituted Systems, involving the precursor proteins and i s o l a t e d mitochondria.
Here we report on the synthesis and tra n s f e r of two mitochondrial membrane proteins, cytochrome c and the carboxyatr-a c t y l o s i d e binding p r o t e i n , the ATP/ADP c a r r i e r of the inner mitochondrial membrane.
I I . RESULTS AND DISCUSSION
A. Cytochrome c
C e l l free homogenates from Neurospora crassa were employed to study biosynthesis and i n t r a c e l l u l a r t r a n s l o c a t i o n of cytochrome c. Antibodies against apocytochrome c and holo-cytochrome c were used to i s o l a t e these proteins by simple im-munoprecipitation. Preexistent proteins were discriminated from in. v i t r o synthesized proteins by dual l a b e l l i n g . For t h i s purpose, Neurospora c e l l s were grown i n 3 5-S-sulfate containing medium and l a b e l l i n g i n the c e l l free homogenate was performed with 3-H-leucine.
Apocytochrome c as well as holocytochrome c were found to be synthesized i n the c e l l free homogenate a f t e r incubation f o r 10 min under appropriate conditions. When pr o t e i n synthesis was then blocked with cycloheximide, the amount of iri v i t r o synthesized apocytochrome c decreased during a further 30 min incubation period. Simultaneously, the amount of newly synthesized holocytochrome c increased (Table I ) . The i d e n t i t i e s of the immunoprecipitated apo- and holocytochrome c were v e r i -f i e d by analysis of the cyanogen bromide fragments.
A c e l l free homogenate l a b e l l e d as described i n Table I was separated by d i f f e r e n t i a l c e n t r i f u g a t i o n into the mitochondrial f r a c t i o n , the microsomal f r a c t i o n and the post-ribosomal supernatant. Immunoprecipitation with antibodies against apocytochrome c and holocytochrome c was c a r r i e d out with a l l f r a c t i o n s .
F i g . 1A and B show that iri v i t r o synthesized apocytochrome c i s present i n the postribosomal supernatant and that i t s amount decreases a f t e r blocking p r o t e i n synthesis. No pree x i s t e n t 3 5-S-labelled holocytochrome c i s p r e c i p i t a t e d by the antibody against apocytochrome c. From the mitochondrial f r a c t i o n anti-apocytochrome c p r e c i p i t a t e d neither 3-H-labelled apocytochrome c nor 3-H- or 3 5-S-labelled holocytochrome c.
Antibodies against holocytochrome c p r e c i p i t a t e d from the postribosomal f r a c t i o n 3 5-S-labelled holocytochrome c, which represents preexistent cytochrome c leaked out from the mitochondria during preparation of the c e l l free homogenate. Also
148 R i c h a r d Z i m m e r m a n n et al.
TABLE I. Immunoprecipitation of Apocytochrome c and of Holocytochrome c from a C e l l Free Homogenate
A c e l l free homogenate was prepared from c e l l s grown i n the presence of 3 5-S-sulfate and incubated at 25 C for 10 min i n the presence of 3-H-leucine. Then cycloheximide (CHI) (100 yg/ ml) was added. One h a l f of the homogenate was withdrawn and kept at 0 C. The other h a l f was further incubated f o r 30 min at 25 C. Then from both portions apocytochrome c and #holocytochrome c were immunoprecipitated. The immunoprecipitates were analysed by SDS Polyacrylamide g e l e l e c t r o p h o r e s i s . 3-H- and 3 5 - S - r a d i o a c t i v i t i e s i n the peak f r a c t i o n were determined.
Antibody 10 min 10 min
+ 30 min CHI (counts x min Ra d i o a c t i v i t y
10 min + 30 min CHI (counts x min
A n t i - 3-H 1826 836 apocytochrome c 35-S 18 24
A n t i - 3-H 260 944 holocytochrome c
35-S 4420 4320
a small amount of in_ v i t r o synthesized holocytochrome c was p r e c i p i t a t e d which increased during incubation i n the presence of cycloheximide (from about 5 to lO% of t o t a l 3-H-holocyto-chrome c i n the homogenate). From the mitochondrial f r a c t i o n the antibody against holocytochrome c brought down preexistent 35-S-holocytochrome c. Also, 3-H-labelled holocytochrome c was found i n the p r e c i p i t a t e . I t s amount increased during i n cubation i n the absence of pro t e i n synthesis (Fig. IC and D).
From the microsomal f r a c t i o n no apocytochrome c could be immunoprecipitated, however a small amount of 3 5-S-holocytochrome c (3-4% of t o t a l ) . Also, 3-H-labelled holocytochrome c was detected i n t h i s f r a c t i o n (10-15% of to t a l ) which did not show a s i g n i f i c a n t increase or decrease during incubation i n the presence of cycloheximide.
These data suggest that apocytochrome c i n the postribosomal supernatant acts as a precursor of holocytochrome c i n the mitochondria.
Transfer of cytochrome c was further i n v e s t i g a t e d i n a re-constituted System, i n which a postribosomal supernatant con-t a i n i n g in_ v i t r o synthesized apocytochrome c was incubated with i s o l a t e d mitochondria from 3 5-S-labelled c e l l s . Table II shows that holocytochrome c appears i n the mitochondria which were r e i s o l a t e d a f t e r incubation. When unl a b e l l e d holo-
P a r t I Elcctrochemical Interactions 149
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FIGURE 1. Immunoprecipitation with antibodies against apocytochrome c from the postribosomal f r a c t i o n and with a n t i bodies against holocytochrome c from the mitochondrial f r a c t i o n . The immunoprecipitates were analysed by SDS gel el e c t r o p h o r e s i s . A,B: Ip with anti-apocytochrome c from the postribosomal supernatant; C,D: Ip with anti-holocytochrome c from the mitochondria; A,C: 10 min incubation; B,D: 10 min incubation plus 30 min i n the presence of cycloheximide.
150 R i c h a r d Z i m m e r m a n n et al.
TABLE I I . Transfer of In V i t r o Synthesized Apocytochrome c i n t o Mitochondria and Conversion to Holocytochrome c
A postmitochondrial homogenate was incubated f o r 10 min with 3-H-leucine and a postribosomal supernatant was prepared. Mitochondria were i s o l a t e d from c e l l s grown i n the presence of 3 5-S-sulfate and resuspended with t h i s postribosomal supernatant. The Suspension was div i d e d i n t o three equal p o r t i o n s . One served as a c o n t r o l , to the second p o r t i o n holocytochrome c (1 nmol/ml) was added, to the t h i r d p o r t i o n apocytochrome c (1 nmol/ml). One h a l f of each p o r t i o n was immediately with-drawn a f t e r resuspension and kept at 0 C, the other h a l f was incubated f o r 30 min at 25 C. Then mitochondria were c o l l e c t e d from each sample by c e n t r i f u g a t i o n and washed with sucrose/ EDTA medium. The mitochondrial p e l l e t s were d i s s o l v e d with T r i t o n X -100 containing buffer and immunoprecipitation with anti-holocytochrome c was c a r r i e d out. The immunoprecipitates were subjected to SDS gel e l e c t r o p h o r e s i s . The 3-H- and 35-S-r a d i o a c t i v i t i e s i n the cytochrome c peaks were determined a f t e r s l i c i n g the gel s .
0 min 30 min Ra d i o a c t i v i t y (counts x min )
Control 3-H 26 440
35-S 3836 4224
+ Holocytochrome c 3-H 72 380
35-S 4016 4112
+ Apocytochrome c 3-H 20 78
35-S 4720 4048
cytochrome c was added to the incubation mixture, no s i g n i f i c a n t change of the amount of holocytochrome c appearing i n the mitochondria was observed. This should be expected since the postribosomal supernatant already contains appreciable amounts of holocytochrome c which leaked out from the mitochondria during c e l l breakage (up to 40% of t o t a l cytochrome c ) . In contrast, a d d i t i o n of apocytochrome c leads to a marked reduction of the amount of holocytochrome c appearing i n the mitochondria.
On the basis of these r e s u l t s the fol l o w i n g mechanism of synthesis and assembly of cytochrome c i s proposed. Apocytochrome c i s synthesized on cytoplasmic ribosomes and released
P a r t 1 E l e c t r o c h e m i c a l I n t e r a c t i o n s 151
i n t o the c y t o s o l . I t seems reasonable to assume that apocytochrome c found i n the postribosomal supernatant i s a c t u a l l y present _in vivo i n the c y t o s o l i n free form. I t can however not d e f i n i t e l y be excluded that i t i s derived from some unknown very f r a g i l e structures which are destroyed during c e l l f r a c -t i o n a t i o n . I t i s f u r t h e r proposed that apocytochrome c d i f f uses to the mitochondrial membrane where the heme group i s c o v a l e n t l y linked to the apoprotein. According to our experi-ments, k i n e t i c data argue against a p o s s i b l e r o l e of microso-mal apo- or holocytochrome c as precursors of mitochondrial holocytochrome c. The conversion of apocytochrome c to holocytochrome c involves a d r a s t i c change i n the conformation of the molecule (Fisher et a l . , 1 9 7 3 ) . I t i s proposed that t h i s conformational change leads to the trapping of newly formed holocytochrome c i n the mitochondrial membrane.
C l e a r l y , our hypothesis r a i s e s a number of questions. They concern the permeability of the outer mitochondrial membrane fo r apocytochrome c, the mechanism by which the heme group i s cov a l e n t l y linked to the apoprotein and the problem, why newly synthesized cytochrome c i s bound to the membrane i n preference to the preexistent cytochrome c which leaked out from the mitochondria.
I t should be pointed out that i n a number of studies on the biogenesis i n r a t l i v e r i t was claimed that holocytochrome c i s synthesized as a whole by the microsomes and eventually t r a n s f e r r e d i n t o mitochondria (for reviews see Sherman and Stewart, 1971; Gonzalez-Cadavid, 1 9 7 4 ) . I t was however not p o s s i b l e i n these studies to discriminate between newly synthesized and preexistent cytochrome c? furthermore, apocytochrome c could not be separately determined. Moreover, the p o s s i b l e precursor r o l e of microsomal cytochrome c was recent-l y r u l e d out by Robbi e t a l . , (1978) (see a l s o Kadenbach, 1 9 7 0 ) .
B. Carboxyatractyloside Binding Protein
The carboxyatractyloside binding p r o t e i n (CAT-protein) or ATP/ADP c a r r i e r represents an i n t e g r a l p r o t e i n of the inner mitochondrial membrane which i s characterized by i t s hydro-phobic properties and an apparent molecular weight of about 32 ,000 (Klingenberg et a l . , 1 9 7 5 ) . I t s synthesis and transfer were studied i n e s s e n t i a l l y the same experimental Systems as described for cytochrome c.
A c e l l free homogenate derived from Neurospora c e l l s grown i n the presence of 3 5-S-sulfate was incubated with 3-H-leucine for 10 min; mitochondria were i s o l a t e d and the CAT-protein was immunoprecipitated. _In v i t r o synthesized CAT-protein was found i n the mitochondria (Fig. 2 A ). In order to detect possible
152 R i c h a r d Z i m m e r m a n n et al.
extramitochondrial precursors, the various s u b c e l l u l a r f r a c t i -ons were subjected to immunoprecipitation, adding mitochondria from unlabelled c e l l s as a source of c a r r i e r p r o t e i n or by double immunoprecipitation. With the ribosomal and microsomal f r a c t i o n s no c l e a r i n d i c a t i o n for the presence of In v i t r o synthesized CAT-protein was found. However, from the p o s t r i b o somal supernatant, 3-H-radioactivity was immunoprecipitated which upon SDS g e l electrophoresis y i e l d e d a peak with ident-i c a l e l e c t r o p h o r e t i c m o b i l i t y as the f u n c t i o n a l CAT-protein. No 35 - S - r a d i o a c t i v i t y was present i n t h i s peak, demonstrating absence of contaminating mitochondrial membrane fragments.
In order to v e r i f y the identy of the 3-H-labelled putat i v e CAT-protein from the postribosomal supernatant, an im-munoprecipitate obtained from t h i s f r a c t i o n was mixed with an immunoprecipitate obtained from mitochondria which were der i v e d from c e l l s l a b e l l e d in_ vivo with 14-C-leucine. The mixture was subjected to cyanogen bromide cleavage and the
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FIGURE 2. Immunoprecipitation of CAT-protein from mitochondria and from a postribosomal p e l l e t . A: Mitochondria; immunoprecipitate obtained d i r e c t l y with antibodies against CAT-protein. B: P e l l e t obtained by 2 h u l t r a c e n t r i f u g a t i o n of a postribosomal supernatant; immunoprecipitate obtained by double p r e c i p i t a t i o n with antibodies against CAT-protein and antibodies against r a b b i t Immunoglobulins.
The immunoprecipitates were analysed by SDS g e l e l e c t r o phoresis .
P a r t I Electrochcmical Interactions 153
r a d i o a c t i v e cleavage products were analysed on phenol/formic a c i d Polyacrylamide gels. The 3-H and 14-C patterns showed s a t i s f a c t o r y coincidence.
Double immunoprecipitation employing anti-rabbit-immuno-globulin-antibodies from sheep lead also to the p r e c i p i t a t i o n of 3-H-labelled CAT-protein from the postribosomal supernatant, however only when T r i t o n X-100 was added. This indicated that the in_ v i t r o synthesized component does not e x i s t i n a free form i n the supernatant, but rather that i t s antigenic s i t e s are not a v a i l a b l e to the antibody. When the postribosomal supernatant was centrifuged f o r 2 h at 165,000 x g, a l l the 3-H-labelled CAT-protein was found i n the p e l l e t (Fig. 2B) .
The question a r i s e s whether the in_ v i t r o synthesized CAT-pr o t e i n i n t h i s f r a c t i o n i s a precursor of the mitochondrial CAT-protein. A c t u a l l y i t was found that the amount of 3-H-l a b e l l e d CAT-protein i n t h i s f r a c t i o n decreased when afte r the 10 min l a b e l l i n g period t r a n s l a t i o n was blocked with c y c l o heximide and the c e l l free homogenate incubated further. On the other hand, 3-H-labelled CAT-protein i n the mitochondrial f r a c t i o n increases under these conditions (Harmey et al.,1977). I t was however not possible to q u a n t i t a t i v e l y c o r r e l a t e these two processes. The main d i f f i c u l t i e s i n these experiments are the exceptional l a b i l i t y of the CAT-protein and the i n a b i l i t y to demonstrate tra n s f e r into f u n c t i o n a l p o s i t i o n s . The newly synthesized p r o t e i n appears to be e a s i l y subject to proteo-l y t i c degradation.
FIGURE 3. Tr a n s l a t i o n of Neurospora CAT-prot e i n i n a ra b b i t r e t i c u -cyte c e l l free System. Coelectrophoresis of immunoprecipitates from the heterologous System (35-S) and from mitochondria l a b e l l e d in_ vivo (3-H).
20 40 Fraction no.
154 R i c h a r d Z i m m e r m a n n et al.
I t i s of i n t e r e s t i n t h i s context whether CAT-protein i s translated as a precursor with i d e n t i c a l or higher molecular weight as compared to the fu n c t i o n a l p r o t e i n i n the mitochondr i a . Poly-A-containing RNA was i s o l a t e d from Neurospora and translat e d i n a r a b b i t r e t i c u l o c y t e l ysate i n the presence of 35-S-methionine. Double immunoprecipitation was c a r r i e d out a f t e r l y s i s with T r i t o n and the r e s u l t i n g p r e e i p i t a t e was mixed with an immunoprecipitate obtained from mitochondria i s o l a t e d from c e l l s l a b e l l e d in_ vivo with 3-H-leucine. The mixture was analysed by SDS gel ele c t r o p h o r e s i s . F i g . 3 shows that the 35-S peak and the 3-H peak with apparent molecular weights of about 32 ,000 coincide. This indicated that CAT-protein i s not translated as a large r molecule, at l e a s t not as a precursor d i f f e r i n g i n i t s apparent molecular weight by more than about 500 from the authentic p r o t e i n .
Further experiments w i l l have to c l a r i f y the precursor r o l e of a postribosomal CAT-protein. As a working hypothesis i t i s proposed that CAT-protein i s not tra n s l o c a t e d as a f r e e l y soluble precursor but rather that binding to l i p i d containing struetures i s involved.
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