COLD SPRING HARBOR SYMPOSIA

ON QUANTITATIVE BIOLOGY

VOLUME XXXVI

COLD SPRING HARBOR SYMPOSIA

ON Q U A N T I T A T I V E BIOLOGY

Founded in 1933

by

REGINALD G. HARRIS

Director of the Biological Laboratory 1924 to 1936

LIST OF PREVIOUS VOLUMES

Volume I (1933) Surface Phenomena Volume II (1934) Aspects of Growth Volume I I I (1935) Photochemical Reactions Volume IV (1936) Excitation Phenomena Volume V (1937) Internal Secretions Volume VI (1938) Protein Chemistry Volume VII (1939) Biological Oxidations Volume VIII (1940) Permeability and the Nature of Cell Membranes Volume IX (1941) Genes and Chromosomes: Structure and Organization Volume X (1942) The Relation of Hormones to Development Volume XI (1946) Heredity and Variation in Microorganisms Volume XII (1947) Nucleic Acids and Nucleoproteins Volume XII I (1948) Biological Applications of Tracer Elements Volume XIV (1949) Amino Acids and Proteins Volume XV (1950) Origin and Evolution of Man Volume XVI (1951) Genes and Mutations Volume XVII (1952) The Neuron Volume XVIII (1953) Viruses Volume XIX (1954) The Mammalian Fetus : Physiological Aspects of Development Volume XX (1955) Population Genetics: The Nature and Causes of Genetic Variability

in Population Volume XXI (1956) Genetic Mechanisms: Structure and Function Volume XXII (1957) Population Studies: Animal Ecology and Demography Volume XXIII (1958) Exchange of Genetic Material: Mechanism and Consequences Volume XXIV (1959) Genetics and Twentieth Century Darwinism Volume XXV (1960) Biological Clocks Volume XXVI (1961) Cellular Regulatory Mechanisms Volume XXVII (1962) Basic Mechanisms in Animal Virus Biology Volume XXVIII (1963) Synthesis and Structure of Macromolecules Volume XXIX (1964) Human Genetics Volume XXX (1965) Sensory Receptors Volume XXXI (1966) The Genetic Code Volume XXXII (1967) Antibodies Volume XXXIII (1968) Replication of DNA in Microorganisms Volume XXXIV (1969) The Mechanism of Protein Synthesis Volume XXXV (1970) Transcription of Genetic Material

STRUCTURE AND FUNCTION OF PROTEINS AT THE

THREE-DIMENSIONAL LEVEL

I

4

!

I

q

COLD SPRING HARBOR SYMPOSIA

ON QUANTITATIVE BIOLOGY

VOLUME XXXVI

STRUCTURE AND FUNCTION

OF PROTEINS AT THE THREE-DIMENSIONAL LEVEL

Q

COLD SPRING HARBOR LABORATORY

1972

COLD SPRING HARBOR SYMPOSIA

ON QUANTITATIVE BIOLOGY

VOLUME XXXVI

�9 1972 by The Cold Spring Harbor Laboratory

All rights reserved

International Standard Book Number: 0-87969-035-6 (clothbound)

Library of Congress Catalog Card Number: 34-8174

Printed in the United States of America

The Symposium Volumes are published by The Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 11724, and may be purchased directly or through booksellers.

Price of Volume 36--$30.00 (inc. postage). Price subject to change without notice.

Front i sp iece : Crystals in the glycolytic pathway. First row: Phosphorlyase, phosphoglucomutase,* hexokinase. Second row: Phosphoglucose isomerase, phosphofructokinase,* atdolase. Th i rd row: Triosephosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase. Four th row: Phospho- glycerate mutase, enolase, pyruvate kinase. *Enzyme~ not yet crystallized in a manner suitable for X-ray diffraction study.

Wil l i am Lawrence B r a g g (1890-1971) was a g rea t sc ien t i s t for over s i x ty years . He looked for s imple answers to i m p o r t a n t p rob lems a n d found more t h a n v i r t ua l l y all his con tempora r i e s . A t his happ i e s t w h e n doing science himself , he neve r rel ished the m a n y a d m i n i s t r a t i v e t a sks t h a t flow to persons of na t i ve good sense. I n h e r e n t l y he was a p r iva t e m a n a n d a t first i n t roduc t i on could seem fo rmidab ly unapp roachab l e . B u t the m o m e n t he l ea rned t h a t a y o u n g e r col league h a d h i t u p o n a new idea, he would become pass iona te ly enve loped in i ts consequences a n d unsel f i sh ly helpful in its fu r the r deve lopmen t . The rap id progress of p ro te in c rys t a l l og raphy over the pa s t two decades b r o u g h t h i m in tense pleasure , pa r t l y because m a n y of the pa r t i c ipan t s were in a real sense his s t uden t s , a n d even more , because of his in tense love for c ry s t a l l og raphy a n d the fac t t h a t once aga in i ts m e t h o d s were the key to ma jo r scientific advances .

He was f o r t u n a t e to be in good hea l t h t h r o u g h o u t m o s t of his life, a n d on ly in his las t yea r was he forced to ser iously res t r ic t h is n a t u r a l e n t h u s i a s m . T h u s it was t h a t he could no t be here to wi tness the m a n y new consequences in the field t h a t he founded du r ing the t r anqu i l Cambr idge s u m m e r of 1912 when he realized t h a t 2d sin 0 = ~ . The dedica t ion of th is v o l u m e to his m e m o r y on ly s l ight ly conveys t h a t v a s t apprec ia t ion wh ich all of us feel for his help in m a k i n g large por t ions of our lives a ve ry exc i t ing adven tu r e . (Photo by Lo t t e Mei tner-Graf , London . )

Symposium Participants ABRAHAM, DONALD J., School of Pharmacy,

University of Pittsburgh, Pittsburgh, Pa. ALDEN, RICHARD A., Dept. of Chemistry, Univer-

sity of California at San Diego, La Jolla, Calif. ALLEN, LELAND C., Dept. of Chemistry, Princeton

University, Princeton, N.J. AMZEL, LEON M., Dept. of Biophysics, The Johns

Hopkins University School of Medicine, Baltimore, Md.

ANDERSON, WAYNE F., Dept. of Molecular Bio- physics and Biochemistry, Yale University, New Haven, Conn.

APPLEBURY, MEREDITHE, Dept. of Biophysics, Bell Telephone Laboratories, Murray Hill, N.J.

ARGOS, PATRICK, Dept. of Physics, Pomona College, Claremont, Calif.

ARNHEIM, NORMAN, Dept. of Biochemistry, State University of New York at Stony Brook, Stony Brook, N.Y.

AULD, DAVID S., Dept. of Biological Chemistry, Harvard Medical School, Boston, Mass.

AVEY, HUGH P., Dept. of Biophysics, The Johns Hopkins University School of Medicine, Baltimore, Md.

BALDWIN, JOYCE, MRC Laboratory of Molecular Biology, Cambridge, England

BALE, JUDITH, Dept. of Chemistry, University of Wisconsin, Madison, Wis.

BANASZAK, LEONARD J., Dept. of Biological Chemistry, Washington University Medical School, St. Louis, Mo.

BEHNKE, WILLIAM D., Dept. of Biological Chem- istry, Harvard Medical School and Biophysics Research Laboratory, Peter Bent Brigham Hospital, Boston, Mass.

BERKOWITZ, STEVEN, Dept. of Biochemistry, New York University, Hackensack, N.J.

BERLINER, LAWRENCE J., Dept. of Chemistry, The Ohio State University, Columbus, Ohio

BERMAN, HELEN, Institute for Cancer Research, Philadelphia, Pa.

BIRKTOFT, JENS J., Dept. of Chemistry, University of California at San Diego, La Jolla, Calif.

BLOO~ER, ANNE C., Dept. of Zoology, Laboratory of Molecular Biophysics, University of Oxford, Oxford, England

BRXND]~N,* CARL-IvAR, Dept. of Chemistry, Agri- cultural College of Sweden, Uppsala, Sweden

BROWN, RODNEY D., III, IBM Thomas J. Watson Research Center, Yorktown Heights, N.Y.

BUEHNER, MANFRED, Dept. of Biological Sciences, Purdue University, Lafayette, Ind.

BUNICK, GERARD, Dept. of Chemistry, University of Pennsylvania, Philadelphia, Pa.

BUTLER, PETER J. G., MRCLaboratory of Molecular Biology, Cambridge, England

CAMERINI-OTERO, RAFAEL D., Dept. of Molecular Biophysics, Public Health Research Institute, New York, N.Y.

CARLSON, WILL~M D., Dept. of Molecular Bio- physics and Biochemistry, Yale University, New Haven, Conn.

CXRTER, CHARLES W., JR., Dept. of Biology, University of California at San Diego, La Jolla, Calif.

CASPAR,* D. L. D., Laboratory of Structural Molecular Biology, Children's Cancer Research Foundation, Boston, Mass.

CLARK, ELOISE E., Dept. of Molecular Biology, Na- tional Science Foundation, Washington, D.C.

COGGINS, JOHN R., Dept. of Biology, Brookhaven National Laboratories, Upton, N.Y.

COHEN, CAROLYN, Laboratory of Structural Molecu- lar Biology, Children's Cancer Research Foun- dation, Boston, Mass.

COHN, MILDRED, Johnson Research Foundation and Dept. of Biophysics and Physical Bio- chemistry, University of Pennsylvania, Phila. delphia, Pa.

CRAVEN, BRYAN M., Dept. of Crystallography,~ University of Pittsburgh, Pittsburgh, Pa.

CROWTHER, R. A., MRC Laboratory of Molecular Biology, Cambridge, England

CUTFIELD, JOHN F., Dept. of Zoology, Laboratory of Molecular Biophysics, Oxford University, Oxford, England

DAHL, LAWRENCE F., Dept. of Chemistry, Uni- versity of Wisconsin, Madison, Wis.

DAVIES, DAVID R., Laboratory of Molecular Biology, National Institutes of Health, Bethesda, Md.

DAY, LOREN A., Dept. of Molecular Biophysics, Public Health Research Institute, New York, N.Y.

DAY, V., Dept. of Chemistry, Massachusetts Institute of Technology, Cambridge, Mass.

DERosIER, DAVID J., Dept. of Chemistry, The University of Texas at Austin, Austin, Tex.

DICKERSON, RICHARD E., Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, Calif.

DOYLE, BARBARA B., Laboratory of Biochemistry, NationalInstitute of Dental Research, National Institutes of Health, Bethesda, Md.

DUNN, MICHAEL, Dept. of Molecular Biochemistry, University of Oregon, Eugene, Ore.

EDELMAN,* GERALD M., Dept. of Biochemistry, The Rockefeller University, New York, N.Y.

EDMUNDSON, ALLEN B., Division of Biological and Medical Research, Argonne National Labora- tory, Argonne, Ill.

* Indicates Chairmen

ix

x SYMPOSIUM PARTICIPANTS

EDSAT.T.,* ffOHN T., Fogarty International Center, National Institutes of Health, Bethesda, Md.

EISENBERG, DAVID, Dept. of Chemistry, University of California at Los Angeles, Los Angeles, Calif.

ENGLANDER, S. WALTER, Dept. of Biochemistry, University of Pennsylvania School of Medicine, Philadelphia, Pa.

EPSTEIN, HENRY F., Laboratory of Chemical Biology, National Institute of Arthritis and Metabolic Diseases, National Institutes of Health, Bethesda, Md.

EVANS, DAVID R., Dept. of Chemistry, Harvard University, Cambridge, Mass.

EVANS, PHILIP R., Department of Zoology, Laboratory of Molecular Biophysics, Oxford University, Oxford, England

FAIRCHILD, BEATRICE M., Dept. of Biological Sciences, Columbia University, New York, N.Y.

FALCoz-KELLY, FRANCOISE, Biochimie Cellulaire, Institut Pasteur, Paris, France

FERSHT, ALAN R., MRC Laboratory of Molecular Biology, Cambridge, England

FLETTERICK, R. J., Dept. of Molecular Biophysics and Biochemistry, Yale University, New Haven, Conn.

FRANKLIN, RICHARD M., Dept. of Molecular Biophysics, The Public Health Research Insti-

tute, New York, N.Y. FREER, STEPHAN T., Dept. of Chemistry, University

of California at San Diego, La Jolla, Calif. GR~ER, JONATHAN, Dept. of Molecular Biophysics

and Biochemistry, Yale University, New Haven, Conn.

GUIDOTTI, GUIDO, Dept. of Biochemistry, Harvard University, Cambridge, Mass.

GUILLORY, RICHARD J., Dept. of Biochemistry and Molecular Biology, Cornell University, Ithaca, N.Y.

GUPTA-BHAYA, PINAKI, Dept. of Chemistry, Colum- bia University, New York, N.Y.

HAL~ORD, S. E., Dept. of Microbiology, Washington University Medical School, St. Louis, Mo.

HAMILTON, WALTER C., Dept. of Chemistry, Brookhaven National Laboratories, Upton, N.Y.

HARDMAN, KARL D., Division of Biological and Medical Research, Argonne National Labora- tory, Argonne, Ill.

HARRISON, STEPHEN C., Laboratory of Structural Molecular Biology, Children's Cancer Research Foundation, Boston, Mass.

HATFIELD, D., IBM Cambridge Scientific, Cam- bridge, Mass.

HENDERSON, RICHARD, Dept. of Chemistry, Yale University, New Haven, Conn.

HENDRICKSON, WAYNE, Laboratory for the Struc- ture of Matter, Naval Research Laboratory, Washington, D.C.

HESS, GEORGE P., Dept. of Biochemistry, Cornell University, Ithaca, N.Y.

HITCHCOCK, SARAH, Dept. of Biology, Brandeis University, Waltham, Mass.

HODGKIN,* DOROTHY C., Dept. of Zoology, Laboratory of Molecular Biophysics, Oxford University, Oxford, England

HOFFMAN, BRIAN M., Dept. of Chemistry, North- western University, Evanston, Ill.

HoL, W. G. J., Laboratory of Structural Chemistry, University of Groningen, Groningen, The Netherlands

HOLMES, KENNETH C., Max-Planck Institut fiir medizinische Forschung Biophysik, Heidel- berg, Germany

HOOGSTEEN, KXRST, Dept. of Biophysics and Pharmacology, Merck Institute for Thera- peutic Research, Rahway, N.J.

HOPFIELD, JOHN J., Dept. of Physics, Princeton University, Princeton, N.J.

HUEER, ROBERT, Max-Planck-Institut fiir Eiweiss und Lederforschung, Munich, Germany

HUXLEY,* HUGH E., Dept. of Biology, Brandeis University, Waltham, Mass.

JACOBSBERG, LAWRENCE, Dept. of Biochemistry and Molecular Biology, Harvard University, Cambridge, Mass.

JANIN, JO~L, Physiologie Microbienne, Institut Pasteur, Paris, France

JARDETZKY, OLEG, Dept. of Pharmacology, Stan- ford University School of Medicine, Stanford, Calif.

JENCKS, WILLIAM P., Graduate Dept. of Bio- chemistry, Brandeis University, Waltham, Mass.

JENSEN, LYLE H., Dept. of Biological Structure, University of Washington, Seattle, Wash.

JOHANSEN, J. T., Dept. of Biological Chemistry, Harvard Medical School, Boston, Mass.

KANNAN, K. K., The Wallenberg Laboratory, University of Uppsala, Uppsala, Sweden

K~RLE, JEROME, Laboratory for the Structure of Matter, Naval Research Laboratory, Washing- ton, D.C.

KATZ, Louis, Dept. of Biological Sciences, Columbia University, New York, N.Y.

KAUZMANN, W., Dept. of Chemistry, Princeton University, Princeton, N.J.

KILMARTIN, JOHN V., MRC Laboratory of Molecular Biology, Cambridge, England

KLOCK, PETER A., Dept. of Biophysics, The Johns Hopkins University, Baltimore, Md.

KLUG, AKRON, MRC Laboratory of Molecular Biology, Cambridge, England

SYMPOSIUM PARTICIPANTS xi

KNOWLES, JEREMY R., The Dyson Perrins Lab- oratory, University of Oxford, Oxford, England

KOENIG, SEYMOUR H., IBM Thomas J. Watson Research Center, Yorktown Heights, N.Y.

KOSHLAND, DANIEL E., JR., Dept. of Biochemistry, University of California at Berkeley, Berkeley, Calif.

KRAUT, JOSEPH, Dept. of Chemistry, University of California at San Diego, La Jolla, Calif.

KRETSINGER, ROBERT H., Dept. of Biology, University of Virginia, Charlottesville, Va.

KUKLA, DIETM~a, Max-Planck-Institut fiir Eiweiss und Lederforschung, Munich, Germany

KUNTZ, I. D., Dept. of Chemistry, Princeton University, Princeton, N.J.

LAKE, JAMES A., The Rockefeller University, New York, N.Y.

LANGRIDGE, ROBERT, Dept. of Biochemical Sciences, Princeton University, Princeton, N.J.

LATTMAN, EATON E., Dept. of Biophysics, The Johns Hopkins University, Baltimore, Md.

LIENHARD, GUSTAV E., Dept. of Chemistry, Harvard University, Cambridge, Mass.

LIPSCOMB,* WILLIAM N., Dept. of Chemistry, Harvard University, Cambridge, Mass.

LovE, WXaNER E., Dept. of Biophysics, The Johns Hopkins University, Baltimore, Md.

LucAs, ROBERT M., Laboratory of Structural Molecular Biology, Children's Cancer Research Foundation, Boston, Mass.

LUDWIG, MARTHA L., Biophysics Research Di- vision, University of Michigan, Ann Arbor, Mich.

MAKINEN, M~VIN W., Laboratory of Biochemistry, National Institute of Dental Research, National Institutes of Health, Bethesda, Md.

MARGOLIASH,* EMANUEL, Dept. of Molecular Biology, Abbott Laboratories, North Chicago, Ill.

MARSH>.LL, VINCENT, Dept. of Biochemistry, University of Illinois, Urbana, Ill.

MATHEWS, F. SCOTT, Dept. of Physiology and Biophysics, Washington University Medical School, St. Louis, Mo.

McCLuRE, RICHARD J., Dept. of Crystallography, University of Pittsburgh, Pittsburgh, Pa.

McDONALD, GEORGE G., Dept. of Biophysics and Biophysical Chemistry, University of Pennsyl- vania, Philadelphia, Pa.

McMuRRAY, C. H., Dept. of Chemistry, Harvard University, Cambridge, Mass.

MEARES, CLAUDE F., Dept. of Chemistry, Stanford University, Stanford, Calif.

MOODY, M. F., Dept. of Cell Biology, The Rocke- feller University, New York, N.Y.

MULVEY, RODERICK, Dept. of Biological Sciences, Columbia University, New York, N.Y.

MfTNCK, ECKARD, Dept. of Physics, University of Illinois, Urbana, Ill.

NORDMAN, C. E., Dept. of Chemistry, University of Michigan, Ann Arbor, Mich.

NORTH, ANTHONY C. T., Dept. of Zoology, Oxford University, Oxford, England

0GATA, RONALD T., Dept. of Chemistry, Stanford University, Stanford, Calif.

0GAWA, SEIJI, Dept. of Biophysics, Bell Telephone Laboratories, Murray Hill, N.J.

PADLAN, EDUARDO A., Dept. of Biophysics, The Johns Hopkins University, Baltimore, Md.

PARRY, DAVID A. D., Laboratory of Structural Molecular Biology, Children's Cancer Research Foundation, Boston, Mass.

PARSONS, STANLEY M., Dept. of Biochemistry, University of California at Berkeley, Berkeley, Calif.

PATEL, DINSI-IAW J., Dept. of Biopolymer Chem- istry, Bell Telephone Laboratories, Murray Hill, N.J.

PERUTZ, MAX F., MRC Laboratory of Molecular Biology, Cambridge, England

PHILLIPS, DAVID C., Dept. of Zoology, Oxford University, Oxford, England

POLJAK, ROBERTO J., Dept. of Biophysics, The Johns Hopkins University School of Medicine, Baltimore, Md.

POWERS, JAMES C., School of Chemistry, Georgia Institute of Technology, Atlanta, Ga.

QuIocHo, F. A., Dept. of Chemistry, Harvard University, Cambridge, Mass.

RAFTERY, MICHAEL A., Dept. of Chemistry, California Institute of Technology, Pasadena, Calif.

REDFIELD, ALFRED G., Dept. of Biochemistry, University of California at Berkeley, Berkeley, Calif.

REEKE, GEORGE N., JR., Dept. of Biochemistry, The Rockefeller University, New York, N.Y.

RICH,* ALEXANDER, Massachusetts Institute of Technology, Cambridge, Mass.

RICHARDS,* FREDERIC M., Dept. of Molecular Biophysics and Biochemistry, Yale University, New Haven, Conn.

RICHARDSON, DAVID C., Dept. of Biochemistry, Duke University, Durham, N.C.

RICHARDSON, JANE S., Dept. of Anatomy, Duke University, Durham, N.C.

RICHMOND, TIMOTHY J., Dept. of Molecular Biophysics and Biochemistry, Yale University, New Haven, Conn.

RIORDAN, JAMES F., Dept. of Biological Chemistry, Harvard Medical School, Boston, Mass.

xii SYMPOSIUM PARTICIPANTS

ROBERTUS, JON D., Dept. of Biology, University of California at San Diego, La Jolla, Calif.

RosE,* IRWIN A., Institute for Cancer Research, Philadelphia, Pa.

ROSENBUSCH, JURG P., Biology Laboratories, Harvard University, Cambridge, Mass.

ROSSMANN, MICHAEL G., Dept. of Biological Sciences, Purdue University, Lafayette, Ind.

RUMBALL, SYLVIA S., Dept. of Chemistry, Cornell University, Ithaca, N.Y.

SALEMME, F. RAYMOND, Dept. of Chemistry, University of California at San Diego, La Jolla, Calif.

SARMA, RAGHUPATHY, Laboratory of Molecular Biology, National Institute of Arthritis and Metabolic Diseases, National Institutes of Health, Bethesda, Md.

SASlSEKHXRAN, V., Dept. of Biochemical Sciences, Princeton University, Princeton, N.J.

SCHECHTER, ALAN N., Laboratory of Chemical Biology, National Institute of Arthritis and Metabolic Diseases, National Institutes of Health, Bethesda, Md.

SCHIFFER, MARIANNE, Dept. of Biological and Medical Research, Argonne National Labora- tory, Argonne, Ill.

SCHILLING, JESSE W., Dept. of Molecular Bio- physics and Biochemistry, Yale University, New Haven, Conn.

SCHOENBORN, BENNO P., Dept. of Biology, Brookhaven National Laboratories, Upton, N.Y.

SCHULTZ, DIANA J., Dept. of Chemistry, Princeton University, Princeton, N.J.

SEARS, DUANE W., Dept. of Biological Sciences, Columbia University, New York, N.Y.

SEEMAN, NADRIAN C., Dept. of Biological Sciences, Columbia University, New York, N.Y.

SH~HN, EZRA, Dept. of Biological Sciences, Hunter College, New York, N.Y.

SHAW, BARBARA R., Dept. of Chemistry, Princeton University, Princeton, N.J.

SHAw, ELLIOTT, Dept. of Biology, Brookhaven National Laboratories, Upton, N.Y.

SHOTTON, DAVID M., Dept. of Biochemistry, University of Bristol, Bristol, England

SHULMAN, ROBERT G., Dept. of Biophysics, Bell Telephone Laboratories, Murray Hill, N.J.

SIMON, GARY L., Dept. of Chemistry, University of Wisconsin, Madison, Wis.

SIMON, SANFORD R., Dept. of Biochemistry, State University of New York at Stony Brook, Stony Brook, N.Y.

SOBELL, HENRY M., Dept. of Chemistry, The University of Rochester, Rochester, N.Y.

STEITZ, THOMAS A., Dept. of Molecular Biophysics

and Biochemistry, Yale University, New Haven, Conn.

STRANDBERG, BROR, The Wallenberg Laboratory, Uppsala University, Uppsala, Sweden

STROUD, ROBERT M., Dept. of Chemistry, California Institute of Technology, Pasadena, Calif.

STRYER, LUBERT, Dept. of Molecular Biophysics and Biochemistry, Yale University, New Haven, Conn.

STUDEBAKER, JOEL F., IBM Thomas J. Watson Research Center, Yorktown Heights, N.Y.

SUSSMAN, JOEL L., Dept. of Biological Sciences, Columbia University, New York, N.Y.

SWAN, IAN D. A., Laboratory of Molecular Biology, National Institute of Arthritis and Metabolic Diseases, National Institutes of Health, Bethesda, Md.

SYKES, BRIAN D., Dept. of Chemistry, Harvard University, Cambridge, Mass.

THOMPSON, ROBERT C., Dept. of Biological Chemistry, Harvard Medical School, Boston, Mass.

VALLEE, BERT L., Biophysics Research Laboratory, Harvard Medical School, Boston, Mass.

VOET, DONALD, Dept. of Chemistry, University of Pennsylvania, Philadelphia, Pa.

WARD, KEITH B., JR., Dept. of Biophysics, The Johns Hopkins University, Baltimore, Md.

WARREN, STEPHEN G., Dept. of Chemistry, Harvard University, Cambridge, Mass.

WATSON, H. C., Dept. of Biochemistry, Bristol University, Bristol, England

WEBER,* KLAUS, Harvard University, Cambridge, Mass.

WEI, C. H., Biology Division, Oak Ridge National Laboratory, Oak Ridge, Tenn.

WESTHEIMER, FRANK H., Dept. of Chemistry, Harvard University, Cambridge, Mass.

WETLAUFER, DONALD B., Dept. of Biochem- istry, University of Minnesota, Minneapolis, Minn.

WHITE, JOHN G., Dept. of Chemistry, Fordham University, Bronx, N.Y.

WIENER, STANLEY L., Medical Research Labora- tory, Long Island Jewish Medical Center, New Hyde Park, N.Y.

WILEY, DON C., Dept. of Biophysics, Harvard University, Cambridge, Mass.

WILLIAMS, R. J. P., Inorganic Chemistry Labora- tory, Wadham College, Oxford, England

WILLIS, ELIZABETH E., Dept. of Chemistry, Princeton University, Princeton, N.J.

WISHNER, BARRY C., Dept. of Biophysics, The Johns Hopkins University, Baltimore, Md.

WODAK, SHOSHANA, Dept. of Biological Sciences, Columbia University, New York, N.Y.

WOOD, MICAL K., General Medical Research,

SYMPOSIUM PARTICIPANTS xiii

Oakland Veterans Administration Hospital, Pittsburgh, Pa.

WOOTTO~, JOHN F., Dept. of Physiology, Bio- chemistry and Pharmacology, NYS Veterinary College, Cornell University, Ithaca, N.Y.

WYCKOFF, HAROLD W., Dept. of Molecular Biophysics and Biochemistry, Yale University,

New Haven, Conn. WYMAN,* J., Istituto Regina Elena, Rome, Italy YANKEELOV, John A., JR., Dept. of Biology,

Brookhaven National Laboratories, Upton, N.Y.

ZIPP, ADAM P., Dept. of Chemistry, Princeton University, Princeton, N.J.

F i r s t r o w : M. A . R a f t e r y , A . R i c h , D . H o d g k i n - - M. F . P e r u t z , D . R . D a v i e s . Second row: H . E . H u x l e y , D . J . D e R o s i e r - - F . S. M a ~ h e w s , R . J . P . W i l l i a m s - - A . C. B l o o m e r , D . C. P h i l l i p s . T h i r d r o w : J . W y m a n , D . L . D . C a s p a r - - B . S t r a n d b e r g , J . T . E d s a l l - - d . B a l d w i n , S. C. H a r r i s o n . F o l t r t h r o w : C . - I . B r a n d 6 n , I . D . A . S w a n - - M. G. R o s s m a n n , A . C. T . N o r t h - - R . H . K r e t s i n g e r , C. C o h e n .

Fi r s t row: R. G. Shu lman , J . J . Hopfield, R. T. Oga ta , A. Klug , P. J . G. Bu t l e r - - D. C. Phi l l ips , A. B. E d m u n d s o n , F. M. R ieha rds .

Second row: S. M. Parsons , R . M. S t roud - - R. J . Po l j ak , H. C. W a t s o n - - S. H. Koenig , L. A. Day . Third row: F. A. Quiocho, J . J . B i rk to f t , R. S a r m a - - R . E. Dickerson, M. A. R a f t e r y - - D. J . DeRos ie r , B. P. Schoenborn. FOurth row: D. C. Wi ley , T. A. Stei tz , J . H. Miller, D. M. Shot ton , H. M. Sobell - - E. Margol iash, J . A. Yankee lov , J r . ,

G. M. E d e l m a n .

F i r s t r o w : W . P . J e n c k s , L . C. A l l e n - - W . C. H a m i l t o n , H . W . W y c k o f f - - W . G . J . H o l , K . K . K a n n a n . S e c o n d r o w : M. S c h i f f e r , C. H . W e i - - P . G u p t a - B h a y a , O . J a r d e t z k y - - D . R . D a v i e s , W . E . L o v e . T h i r d r o w : E . S h a w , J . R . C o g g i n s - - K . W e b e r . - - R . H e n d e r s o n , M. Z w i c k . F o u r t h r o w : J . V . K i l m a r t i n , R . T . O g a t a - - L . H . J e n s e n , E . E . L a t t m a n .

FOREWORD

Protein structure analysis is now a field very different from its counterpart of only a few years ago. Then, most inferences about what proteins look like at the three- dimensional level were indirect and most speculations about how they acted were quickly forgotten. But spectacular advances in protein crystallography made during this past decade now present us with precise atomic coordinates for a number of important proteins, thereby allowing the formulation of detailed mechanisms for their manner of function at the molecular level In this work, important contributions have been made by people of highly diverse backgrounds. Mathematicians, physicists, physical chemists, organic chemists, and biochemists have all played indispensible roles. This year's Symposium thus seemed a good time to bring together these various people to talk about where protein work now stands and to consider which proteins are next ripe for full understanding.

Drs. Aaron Klug, William Lipscomb, Max Perutz, David Phillips, and Fred Richards were a tremendous help in drawing up this program and we wish to thank them for their many suggestions. Our appreciation extends to Drs. Sanford Simon and William Lipscomb who generously supplied their special projection equipment used during the Symposium. We wish to express our gratitude to Dr. Fred Richards, without whose guidance the task of publishing the two-color stereo diagrams would not have been possible; to Drs. Masden and James (Alberta) for pro- viding photomicrographs of crystals of phosphorylase; and Dr. Steitz (Yale) and Mr. Eagles (Oxford) for providing crystals of hexokinase and aldolase for photo- micrography. The remaining crystals shown in the frontispiece were kindly provided by Dr. H. C. Watson and members of the Biochemistry Department, Bristol, who were also responsible for preparing the necessary photomicrographs and assembly of the collage.

J. D. Watson

Contents

Dedication . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Symposium Participants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Foreword . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

vii xiii xix

GENERAL CONSIDERATION OF PROTEIN STRUCTURE AND CATALYSIS

Structure-Reactivity Correlations and General Acid-Base Catalysis in Enzymic Transacylation Reactions W. P. Jencks . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

The Importance of Orientation Factors in Enzymatic Reactions D . E . Kosh- land, Jr., K. W. Carraway, G. `4. Dafforn, J. D. Gass, and D. R. Storm . . .

Views on Approximation, Orbital Steering, and Enzymatic and Model Reactions T. C. Bruice . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

The Role of Distortion in the Lysozyme Mechanism B . D . Sykes, S. L. Part, and D. Dolphin . . . . . . . . . . . . . . . . . . . . . . . . . . . : . . . . . . . . . . . . . . . . . . . . . . .

Protein Structure, Ribonuclease-S and Nucleotide Interactions F . M . Richards, H. W. Wyckoff, W. D. Carlson, N. M. ,411ewell, B. Lee, and Y. Mitsui . . .

Enzymatic Catalysis and the Transition State Theory of Reaction Rates: Transition State Analogs G. E. Lienhard, I. I. Secemski, K. A. Koehler, and R. iV. Lindquist . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

The Entatic State R. J. P. Williams . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

1

13

21

29

35

45 53

PROTEASES: STRUCTURE, CATALYSIS, INHIBITION

~-Chymotrypsin: What Can We Learn about Catalysis from X-Ray Diffraction ? R. Henderson, C. S. Wright, G. P. Hess, and D. M. Blow . . . . . . . . . . . . . . .

Conformational Equilibria and the Salt Bridge in Chymotrypsin A . R . Fersht . Spectrophotometric and Structural Evidence as to the Mechanism of Protease

Catalysis at Chemical Bonding Resolution S. .4. Bernhard and S.-J. Lau .. The Stereochemistry of Substrate Binding to Chymotrypsin Ay D . M . Segal,

G. H. Cohen, D. R. Davies, J. C. Powers, and P. E. Wilcox . . . . . . . . . . . . . Conformational Changes and Inhibitor Binding at the Active Site of Elastase

D. M. Shotton, iv. J. White, and H. C. Watson . . . . . . . . . . . . . . . . . . . . . . . . A Comparison of the Three-dimensional Structures of Subtilisin BPN' and

Subtilisin Novo J. Drenth, W. G. J. Hol, J. N. Jansonius, and R. Koekoek . The Aromatic Substrate Binding Site in Subtilisin BPN' and Its Resemblance

to Chymotrypsin J. Kraut, J. D. Robertus, J. J. Birktoft, R. A. Alden, P. E. Wilcox, and J. C. Powers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

The Crystal and Molecular Structure of DIP-inhibited Bovine Trypsin at 2.7/~ Resolution R. M. Stroud, L. M. Kay, and R. E. Dickerson . . . . . . . . . . . .

Pancreatic Trypsin Inhibitor (Kunitz) Part I. Structure and Function R. Huber, D. Kukla, `4. Ruhlmann, and W.

Steigemann . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

xxi

63 71

75

85

91

107

117

125

141

xxii CONTENTS

Part II. Complexes with Proteinases A. Riihlmann, H. J. Schramm, D. KuIda, and R. Huber . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 148

ENZYMES IN THE GLYCOLYTIC PATHWAY

Crystallographic Studies of Chicken Triose Phosphate Isomerase D . W . Banner, A. C. Bloomer, G. A. Petsko, D. C. Phillips, and C. I. Pogson . . . . . . . . . .

Triosephosphate Isomerase: Isotope Studies on the Mechanistic Pathway J. R. Knowles, P. F. Leadlay, and S. G. Maister . . . . . . . . . . . . . . . . . . . . . . . .

X-Ray Diffraction Studies on Enzymes in the Glycolytic Pathway J. W. Campbell, E. Duee, G. Hodgson, W. D. Mercer, D. K. Stammers, P. L. Wendell, H. Muirhead, and H. C. Watso~ . . . . . . . . . . . . . . . . . . . . . . . . . . . .

151

157

165

DEHYDROGENASES

Structural Studies on Heart Muscle Malate Dehydrogenases D. Tsernoglou, E. Hill, and L. J. Banaszak . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Structural Constraints on Possible Mechanisms of Lactate Dehydrogenase as Shown by High Resolution Studies of the Apoenzyme and a Variety of Enzyme Complexes M. G. Rossmann, M. J. Adams, M. Buehner, G. C. Ford, M. L. Hackert, P. J. Lentz, Jr., A. McPherson, Jr., R. W. Schevitz, and I. E. Smiley . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Relaxation Studies on an Allosteric Enzyme: Aspartokinase I-Homoserine Dehydrogenase I J. Janin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

A Low Resolution Electron-Density Map of Lipoyl Transsuccinylase, the Core of the ~-Ketoglutarate Dehydrogenase Complex D . J . DeRosier and R. M. Oliver . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

171

179

193

199

MUSCLE PROTEINS

Tropomyosin Crystal Dynamics C. Cohen, D. L. D. Caspar, D. A. D. Parry, and R. M. Lucas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

The Structure of a Calcium-binding Protein from Carp Muscle R . H . Kretsinger, C. E. Nocbolds, C. J. Coffee, and R. A. Bradshaw . . . . . . . . . . . . . . . . . . . . .

205

217

NEW PROTEIN STRUCTURES AND PROGRESS REPORTS

Crystal Structure of Human Erythrocyte Carbonic Anhydrase C. VI. The Three- dimensional Structure at High Resolution in Relation to Other Mammalian Carbonic Anhydrases K. K. Kannan, A. Liljas, I. Waara, P.-C. Bergstdn, S. L~ivgren, B. Strandberg, U. Bengtsson, U. Carlbom, K. Fridborg, L. Jdrup, and M. Peter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

The Crystal Structure of Rhombohedral 2 Zinc Insulin T . L . BlundeU, J. F. Cutfield, E. J. Dodson, G. G. Dodson, D. C. Hodglcin, and D. A. Mercola

Some Aspects of the Structure of Staphylococcal Nuclease Part I. Crystallographic Studies F. A. Cotton, C. J. Bier, V. W. Day, E. E.

Hazen, Jr., and S. JLarsen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Part II. Studies in Solution C. B. Anfinsen, A. N. Schechter, and H. Taniuchi

221

233

243 249

CONTENTS Txill

Tentative Sequential Model for the Unfolding and Refolding of Staphylococcal Nuclease at High pH O. Jardetzky, J. L. Markley, H. Thielmann, Y. Arata, and M. N. Williams . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Concerning the Stereochemistry of Actinomycin Binding to DNA: An Actino- mycin-Deoxyguanosine Crystalline Complex H. M. Sobell, S. C. Jain, T. D. Sakore, G. Ponticello, and C. E. Nordman . . . . . . . . . . . . . . . . . . . . . . . . .

X-Ray Crystallographic Studies of Concanavalin A K. D. Hardman, M. K. Wood, M. Schiffer, A. B. Edmundson, and C. F. Ainsworth . . . . . . . . . . . . .

The Structure of Concanavalin A at 4 A Resolution G.N. Reeke, J . W. Becker, and F. A. Quiocho . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

The 5.5/~ Resolution Structure of the Regulatory Enzyme, Aspartate Trans- carbamylase D. C. Wiley, D. R. Evans, S. G. Warren, C. H. McMurray, B. F. P. Edwards, W. A. Franks, and W. N. Lipscomb . . . . . . . . . . . . . . . . .

Molecular Symmetry and Crystal Packing of E. coli Glutamine Synthetase D. Eisenberg, E. G. Heidner, P. Goodkin, M. N. Dastoor, B. H. Weber, F. Wedler, and J. D. Bell . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

257

263

271

277

285

291

HEMOGLOBINS: MUTANTS, DERIVATIVES, AND COMPLEXES

Stereochemistry of Cooperative Effects in Hemoglobin M. F. Perutz and L. F. TenEyck . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

The Effect of Removal of C-terminal Residues on Cooperative Interactions in Hemoglobin J. V. Kilmartin and J. A. Hewitt . . . . . . . . . . . . . . . . . . . . . . .

Three-dimensional Structure of Abnormal Mutant Human Hemoglobins J. Greet . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

The Binding of a Spin-labeled Triphosphate to Hemoglobin R . T . Ogata and H. M. McConnell . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

An Allosteric Model of Hemoglobin R. G. Shulman, S. Ogawa, and J. J. H opfield . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Coboglobins: Cobalt Substitution and the Nature of the Prosthetic Group-- Apoprotein Interaction in Hemoglobin and Myoglobin B . M . Hoffman, C. A. Spilburg, and D. H. Petering . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

The Structures of Lamprey and Bloodworm Hemoglobins in Relation to Their Evolution and Function W. E. Love, P. A. Klock, E. E. Lattman, E. A. Padlan, K. B. Ward, Jr., and W. A. Hendrickson . . . . . . . . . . . . . . . . . . . . .

295

311

315

325

337

343

349

IRON-CONTAINING PROTEINS OTHER THAN HEMOGLOBIN

The Structure of a Non-Heme Iron Protein: Rubredoxin at 1.5 A Resolution K. D. Watenpaugh, L. C. Sieker, J . R. Herriott, and L. H. Jensen . . . . . . .

The Structure of a Clostridial Flavodoxin, an Electron-transferring Flavoprotein. III. An Interpretation of an Electron-Density Map at a Nominal Resolution of 3.25 A M. L. Ludwig, R. D. Andersen, P. A. Apgar, R. M. Burnett, M. E. LeQuesne, and S. G. Mayhew . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Structure of the Iron-Sulfur Cluster in the Chromatium Iron Protein at 2.25/~ Resolution C. W. Carter, Jr., S. T. Freer, Ng. H. Xuong, R. A. Alden, and J. Kraut . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

The Structure of Cytochrome b 5 at 2.0 A Resolution F . S . Mathews, P. Argos, and M. Levine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Conformational Changes upon Reduction of Cytochrome c T. Takano, R. Swanson, O. B. Kallai, and R. E. Dickerson . . . . . . . . . . . . . . . . . . . . . . . . . .

Pulsed NMR Study of the Structure of Cytochrome c A . G . Redfield and R. K. Gupta . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

359

369

381

387

397

405

xxiv CONTENTS

IMMUNOGLOBULIN STRUCTURE: IgG AND FRAGMENTS

Crystal Structure of an Immunoglobulin Molecule by X-Ray Diffraction and Electron Microscopy V. R. Sarma, D. R. Davies, L. W. Labaw, E. W. Silverton and W. D. Terry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

X-Ray Crystallographic Studies of the Fab and Fc Fragments of Human Myeloma Immunoglobulins R. J. Poljak, L. M. Amzel, H. P. Avey, L. N. Becka, D. J. Goldstein, and R. L. Humphrey . . . . . . . . . . . . . . . . . . . . . . . . . .

Crystallographic Studies of an IgG Immunoglobulin and the Bence-Jones Protein from One Patient A. B. Edmundson, M. Schiller, M. K. Wood, K. D. Hardman, K. 1~. Ely, and C. F. Ainsworth . . . . . . . . . . . . . . . . . . . . . . . .

413

421

427

STRUCTURE OF VIRUSES

An Electron-Density Map of Tobacco Mosaic Virus at 10 .~ Resolution A.N. Barrett, J . Barrington Leigh, K. C. Holmes, A. Klug, R. Leberman, E. Mandelkow, and P. Von Sengbusch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

The Disk of TMV Protein and Its Relation to the Helical and Other Modes of Aggregation A. Klug and A. C. H. Durham . . . . . . . . . . . . . . . . . . . . . . . . .

The Mechanism and Control of the Assembly of Tobacco Mosaic Virus from Its RNA and Protein Disks P . J . G . Butler . . . . . . . . . . . . . . . . . . . . . . . . . . . .

X-Ray Diffraction Studies of the Structure of Satellite Tobacco Necrosis Virus K. Akervall, B. Strandberg, M. G. Rossmann, U. Bengtsson, K. Fridborg, H. Johannisen, K. K. Kannan, S. Lovgren, G. Pete f , B. Oberg, D. Eaker, S. Hjertdn, L. Ryden, and I. Moking . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Three-dimensional Image Reconstructions of Some Small Spherical Viruses R. A. Crowther and L. A. Amos . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Structure of Tomato Bushy Stunt Virus: Three-dimensional X-Ray Diffraction Analysis at 30 A Resolution S .C . Harrison . . . . . . . . . . . . . . . . . . . . . . . . .

Structural Studies on the Adenovirus Hexon R . M . Franklin, S. C. Harrison, U. Pettersson, C.-I. Br~inddn, P.-E. Werner, and L. Philipson . . . . . . . . . . .

433

449

461

469

489

495

503

SPECTROSCOPY AND SPECTROSCOPIC PROBES

The Study of Biological Macromolecules Using Perturbed Angular Correlations of Gamma Radiation C. F. Meares and D. G. Westmoreland . . . . . . . . . . .

Speetro-chemical Probes for Protein Conformation and Function B . L . Vallee, J. F. Riordan, J. T. Johansen, and D. M. Livingston . . . . . . . . . . . . . . . . . . .

Mapping Active Sites of Phosphoryl-transferring Enzymes by Magnetic Resonance Methods M. Cohn, J. S. Leigh, Jr., and G. H. Reed . . . . . . . . .

Use of 19F-Nuclear Magnetic Resonance Spectroscopy for Detection of Protein Conformation Changes: Application to Lysozyme, Ribonuclease, and Hemoglobin M. A. Raftery, W. H. Huestis, and F. Millett . . . . . . . . . . . .

On the Interpretation of Solvent Proton Magnetic Relaxation Data with Par- ticular Application to the Structure of the Active Site of Mn-Carboxy- peptidase A S. H. Koenig, R. D. Brown, and J. Studebaker . . . . . . . . . . . .

X-Ray Diffraction and Nuclear Magnetic Resonance Dispersion Studies on Derivatives of Carboxypeptidase A F. A. Quiocho, P. H. Bethge, W. N. Lipscomb, J. F. Studebaker, R. D. Brown, and S. H. Koenig . . . . . . . . . . . .

511

517

533

541

551

561

CONTENTS xxv

TECHNIQUES IN DIFFRACTION AND MODEL BUILDING

A Neutron Diffraction Analysis of Myoglobin. III. Hydrogen-Deuterium Bonding in Side Chains B . P . Schoenborn . . . . . . . . . . . . . . . . . . . . . . . . . . .

The Use of a Computer-controlled Display System in the Study of Molecular Conformations C. D. Barry and A. C. T. North . . . . . . . . . . . . . . . . . . . . . .

Construction of Space-filling Models of Proteins Using Dihedral Angles J . A . Yankeelov, Jr., and J. R. Coggins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

569

577

585

SUMMARY

Protein Crystallography 1971 : Coming of Age D . C . Phillips . . . . . . . . . . . . . . 589

Appendix: Stereo Plates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Subject Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Name Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

593

633

639