Citric Acid Cycle

778 Pages·1969·13.3 MB·English

by  John M. Lowenstein (Eds.)

Contributors to Volume XIII
Article numbers are shown in parentheses followlng the names of contributors.
Affiliations listed are current.
S. J. AJL (55), Department o Bio- vania School o Medicine, Philadelphia,
chemistry, Albert Einstein Medical Pennsylvania
Center, Philadelphia, Pennsylvania Boa B. NANAHCUB (30), Department o
YCNAN W. KCOOLA (61), Sloan Kettering Cell Physiology, University o Cali-
Institute or Cancer Research, New ornia, Berkeley, ainroilaC
York, New York J. L. ChNovAs (45), Instituto de BioIogia
S. H. G. NELLA (33, 41), Department o ,ralulleC C ~.I.C., Madrid, Spain
Biochemistry, Albany Medical College, Louis N. CECi (70, 78), The Murray
Albany, New York Strassman Memorial Laboratory, De-
LmNAD I. NONRA (30), Department o partment o Microbiology, Albert
Cell Physiology, University o Cali- Einstein Medical Center, Philadelphia,
,ainro Berkeley, ainroilaC Pennsylvania
R. E. NEVaAB (7), Department o Bio- NAMONUS AHC (13), Department o Bio-
chemistry, University o Wisconsin, logical and Medical Sciences, Brown
Madison, Wisconsin University, Providence, Rhode Island
H. A. REKRAB (50, ,15 52), Department H. C. CHANa (42), Biochemistry De-
o Biochemistry, University o CaIi- partment, Central Research, Shulton,
ornia, Berkeley, ainroilaC Inc., Cliton, New Jersey
EHSOM NAMXZNEB (22), Laboratory o J. F. A. CHASE (60), Department o
Biochemistry, University o Cam-
cinagrO and Biological Chemistry, The
bridge, Cambridge, England
Hebrew University o Jerusalem, Jeru-
salem, Israel W. W. )INALELC (7), Department o Bio-
chemistry, University o Wisconsin,
ARDNENANJ K. EEJRAHCATTAHB (78), The
Madison, Wisconsin
Murray Slrassman Memorial Labora-
R. A. Cook (9), Department o Bio-
tory, Department o Microbiology,
chemistry, University o Caliornia,
Albert Einstein Medical Center, Phila-
Berkeley, California
delphia, Pennsylvania
R. A. REPOOC (48), Department o Bio-
NATIE NIGOB (4), Department o Plant
chemistry, University o Leicester,
Biochemistry, University o ,alnroilaC
Leicester, England
Los Angeles, ainroilaC
ARABRAB E. CORKEr (65), Graduate De-
P. D. REROB (14), Molecular Biology partment o Biochemistry, ~.iednarB
Institute, University o Caliornia, Los
University, Waltham, Massachusetts
Angeles, ainroilaC
G. F. Cox (10), School o Biological
HPLAR A. WAHSDaRB (17), Department Sciences, University o East Anglia,
o Biochemistry, University o Wash- Norwich, England
ington School o Medicine, Seattle, S. YELOAD (28, 67), Department o Bio-
Washington
chemistry, College o Biological Sci-
W. A. REGDIRB (14), Department o Bio- ences, University o/ Minnesota, St.
chemistry, University o Alberta, Ed- Paul, Minnesota
monton, Alberta, Canada DIVAD D. SEIVAD (26), School o Biologi-
DLORAH J. BRIaHT (53), Department o lac Sciences, University o East Anglia,
Biochemistry, University o Pennsyl- Norwich, England
vi CONTRIBUTORS TO VOLUME XIII
HTXDUJ J. DAvls (47), Department o chemistry, Dental School, Osaka Uni-
Psychiatry, Stanord Medical Center, versity, Osaka, Japan
Pals Alto, Caliornia BmGIT NOSROCAJ (36), Department o
D. V. NAINATRAVRED (16), Institute or Biochemistry, Case Western Reserve
Enzyme Research, University o Wis- University School o Medicine, Clevc-
consin, Madison, nis~rocsiW land, Ohio
TREBOR J. DUMMEL (79), Department o WM. P. JENCKS (15), Graduate Depart-
Pharmacology, University o CaliIornia ment o Biochemistry, Brandeis Uni-
School o Medicine, San Francisco, versity, Waltham, Massachusetts
ainroilaC HAKUJI ARUSTAK (76), Faculty o Sci-
LEINAD EUQRUOPU)I (20), Deparlment o ence, Osaka University, Osaka, Japan
Pharmacology, University o ,ainroilaC OTIHSOY KAzmo (31), Department o
San Francisco Medical Center, School Chemistry, Institute o Medical Sci-
o Medicine, San Francisco, Caliornia ence, University o Tokyo, Minatoku,
R. L. YA)mETS^E (43), Pharmacia Fine Tokyo, Japan
Chemicals, Inc., Piscataway, New R. W. REYEMRELLEK (33, 35), Depart-
Jersey ment o Medicine, Case Western Re-
AHSAS DRALONE (18, ,12 74), Biochemistry serve University School o Medicine,
Department, Albert Einstein College Cleveland, Ohio
o Medicine, Bronx, New York OEL KESNER (62), Department o Bio-
B. aELSNAF (6), The Institute or Cancer chemistry, College o Medicine, Down-
Research, Philadelphia, Pennsylvania state Medical Center, State University
P. B. GA~HD (2, 72), Department o o New York, Brooklyn, New York
Biochemistry, The Medical School, G. EIRRAB grrTO (19, 25), Department o
Bristol, England Chemistry, Clayton Foundation Bio-
ADNERB I. G~RWIH (36), Department o chemical Institute, University o Texas,
Biochemistry, Case Western Reserve Austin, Texas
University School o Medicine, Cleve- TUMLEH R. HCSZTOLK (59), Boehringer
land, Ohio Mannheim Corp., New York, New
HTENNEK R. HANSON (74), The Con- York
necticut Agricultural Experiment Sta- H. L. OREDNROK (45, 48), Department o
tion, New Haven, Connecticut Biochemistry, University o Leicester,
Lovm B. HSREH (15), Laboratory o Bio- Leicester, England
chemistry, National Heart Institute, TSENRE KUN (20, 79), Department o!
National Institutes o Health, Be- Pharmacology, University o California,
thesda, Maryland San Francisco Medical Center, School
TREBOR L. HILL (17), Department o Bio- o Medicine, San Francisco, Caliornia
chemistry, Duke University, Durham, M. DANIEL LANE (42, 43), Department
North Carolina o Biochemistry, New York University
.vLwavM W. HSnNO (53), Department o School o Medicine, New York, New
Biophysics, University o Chicago, York
Chicago, Illinois YRNEI-I A. YDRAL (37, 49), Institute ro
R. Y. Hsv (37), Department o Bio- Enzyme Research, The University o
chemistry and Microbiology, College Wisconsin, Madison, Wisconsin
o Agriculture and Environmental Sci- DLANOD J. EUGITRAL (54), Department o
ence, Rutgers, The State University, Biochemistry, Louisiana State Uni-
New Brunswick, New Jersey versity, Baton Rouge, anai.~iuoL
KuN-YEN I-IuANO (64), Department o Y. S. LEwis (77), Central Food Techno-
Microbiology, Naval Medical Research logical Research Institute, Mysore,
Institute, Bethesda, Maryland India
OEDIH INOUE (27), Department o BIO- W. D. LooM1S (73), Department o Bio-
CONTRIBUTORS TO VOLUME XIIl vii
chemistry and Biophysics, Oregon State versity School o Medicine, Cleveland,
University, Corvallis, Oregon Ohio
JOH~ M. NaFTSNEWOL (6, 66), Graduate R. NIVRAP (3), Biochemistry Division,
Department So Biochemistry, Brandeis Department o Biochemistry, Univer-
University, Waltham, Massachusetts sity o Caliornia, Los Angeles, CaIi-
F. A. McELRoY (71), Department o Bio- ainroJ
chemistry, University o Toronto, P. J. R. YELREKCAZIHP (23), Nuffeld De-
Toronto, Canada partment o Clinical Biochemistry,
ECURB A. NEDDAFcM (29), Department University o Oxord, Oxord, England
o Chemistry, Washington State Uni- G. W. E. TUALP (8), Department o Bio-
versity, Pullman, Washington chemistry, R~tgers Medical School,
P. AREAM (44), Laboratory o Molecu- New Brunswick, New Jersey
lar Biology, University o Wisconsin, J. R. ELYAUQ (46, 57), Department o
Madison, Wisconsin Microbiology, The University o She-
H. AMAYURAM (43), Research Institute, field, Sheffield, England
St. Joseph Hospital, Lancaster, Penn- R. RABIN (55), Department o Bio-
sylvania chemistry, Albert Einstein Medical
IHSRAJAR REDMUZAM (32, 34), Depart- Center, Philadelphia, Pennsylvania
ment o Biochemistry, All-India Insti- R. F. YELAMAR (14), Department o
tute o Medical Sciences, New Delhi, Microbiology, Indiana University,
India Bloomington, Indiana
TREBOR S. RELLIM (42), Department o RETSEL J. REED (12), Department o
Biochemistry, New York University Chemistry, Clayton Foundation Bio-
School o Medicine, New York, New chemical Institute, University o Texas,
York Austin, Texas
DIRAB B. EEJREHKUM (12), Corporate I~. C. SEVEER (55), Department o Bio-
Research, Tenneco Chemicals, Inc., chemistry, Albert Einstein Medical
Piscataway, New Jersey Center, Philadelphia, Pennsylvania
DRAWDE RELYWTNUM (62), Department D. R. IUANAS (11), Department o -OIB
o/ Biochemistry, College o Medicine, energetic Research, Retina Foundation,
Downstate Medical Center State Uni- Boston, Massachusetts
versity o New York, Brooklyn, New B. D. LAWNAS (9, 44), Department o
York Microbiology, University o Manitoba,
MAILLIW H. YEHPRUM (25), Department Winnipeg, Canada
~o Pediatrics, State University o New M. C. SCRU~ro~ (38), Department o
York at Bufalo Medical School, Bu- Biochemistry, Rutgers University
/alo, New York School o Medicine, New Brunswick,
MAILLIW F. SREYM (64), Department o New Jersey
Microbiology, University o Maryland W. TREBUES (40), Institut iir Vegetative
Medical School, Baltimore, Maryland Physiologie der Universit~it, Chemisch-
RETXED B. PORHTRON (36), Department Physiologisches Institut der Stadt
o/ Biochemistry, Case Western Reserve Frankurt, Frank~lrt/Main, Germany
University School o Medicine, Cleve- D. DREHPEHS (2), Shell Centre, London,
land, Ohio England
OREVES AOHCO (34), Department o Bio- SIWEL LEGEIS (18), Biochemistry Depart-
chemistry, New York University, ment, Albert Einstein College o Medi-
School o Medicine, New York, New cine, Bronx, New York
York LUAP A. EaERS (1), Veterans Administra-
M. R. DETSMLO (38), Department o Bio- tion Hospital. ,sald~D Texas
chemistry, Case Western Reserve Uni- YARRUM "NAMSSARTS (70), $ Department
* Deceased.
Vlll o,° SROTUBIRTNOC OT VOLUME XIII
o Microbiology, Albert Einstein Medi- serve University School o/ Medicine,
cal Center, Philadelphia, Pennsylvania Cleveland, Ohio
FuJIo SuzuxI (27), Department o Bio- W. S. RENEOEW (55), tnemtrapeD o/
chemistry, Osaka University Dental Biochemistry, Albert Einstein Medical
School, Osaka, Japan Center, Philadelphia, Pennsylvania
ORIHSOY ADEKAT (27), tnemtrapeD fo H. REKCIEW (40), Institut fiir Vegetative
Biochemistry, Osaka University Dental Physiologie der Universittit, Chemisch-
School, Osaka, Japan Physiologisches In~titut der Stadt
V. W. NOSPMOHT (7), Department o Frankurt, Frankurt/Main, Germany
Biochemistry, University o Wisconsin, P. D. J. NAMZVIEW (5, 56), Department
Madison, Wisconsin o Biochemistry, University o Leices-
BERNADINE TOLBERT (39), Department o ter, Leicester, England
Biochemistry, Case Western Reserve SEMAJ M. DaALLIW (47), Department o
University School o Medicine, Cleve- Biochemistry, Case Western Reserve
land, Ohio University School o Medicine, Cleve-
P. K. SBBUT (72), Department o Bio- land, Ohio
chemistry, University o Cambridge,
G. R. WILL,MS (71), Department o
Cambridge, England
Biochemistry, University o Toronto.
YNOHTNA F. TvccI (78), The Murray
Toronto, Canada
Strassman Memorial Laboratory, De-
AXNICaIV R. SMAILLIW (54), Department
partment o Microbiology, Albert Ein-
of Biochemistry, Louisiana State Uni-
stein Medical Center, Philadelphia,
versity, Baton Rouge, Louisiana
Pennsylvania
JOHN R. NOSMAILLIW (65), Johnson Re-
M. F. RETTU (3~, 39), Department of
hcraes Foundation, Department o Bio-
Biochemistry, Case Western Reserve
physics and Physical Biochemistry,
University School o Medicine, Cleve-
University o Pennsylvania Medical
land, Ohio
School, Philadelphia, Pennsylvania
C. REGE-~V (16, 69), Department o Bio-
chemistry, Agricultural University, HAEaNI) G. DOOW (33, ,53 36, 47), De-
partment o Biochemistry, Case West-
Wageningen, The Netherlands
ern Reserve University School o
H. B. YRZKCXV (75), The Connecticut
Medicine, Cleveland, Ohio
Agricultural Experiment Station, New
Haven, Connecticut AKmA ADIHSOY (24), Medical Genetics,
R. W. VoN KoR~ (63, 68), Friends o Department o Medicine, University
Psychiatric Research, Incorporated, o Washington School o Medicine,
Baltimore, Maryland Seattle, Washington
EUHTaA W^LL^CZ (4), Department o M. R. GNUOY (39), Department o Bio-
Plant Biochemistry, University o Cali- chemistry, Case Western Reserve Uni-
ornia, Los Angeles, California versity School o Medicine, Cleveland,
C. C. W^N(~ (51), Department o Bio- oihO
chemistry, University o California, W. P. EEKAMELYEZ (16, 69), Laboratory
Berkeley, ainroilaC of Biochemistry, B. C. P. Jansen Insti-
EILSEL .W ,RETSBEW .RJ (58), Department tute, University o Amsterdam, Am-
o Pharmacology, Case Western Re- sterdam, The Netherlands
Preface
The citric acid cycle represents the terminal stage for the oxidation of
the major foodstuffs and energy stores in many organisms. In addition,
the cycle plays an important part in the synthesis of many cell constit-
uents from simple precursors. This volume of "Methods in Enzymology"
deals w.ith the reactions of the citric acid cycle and with a number of
reactions leading to and from the cycle. In deciding what to include in
the present volume, boundaries had to be drawn between various areas
of metabolism and the citric acid cycle. These boundaries are of necessity
somewhat arbitrary. The aspartase reactiofi is included. On the other
hand, enzymes which might have been included here, such as certain
transaminases and glutamate dehydrogenase, will be covered in a future
volume devoted to amino acid metabolism. A few of the methods which
are included should logically have appeared in earlier volumes. These are
warranted by the importance of the topic and the time it would take for
them to appear in future revisions of recent volumes.
The help and cooperation of the contributors is greatfully acknowl-
edged. I am indebted to Drs. H. L. Kornberg, H. A. Lardy, and H. Vickery
for useful suggestions, to Mrs. Janice Bright, Miss Geraldine Conner, and
Miss Kathryn Rader for their skilled and patient secretarial help, and to
the Staff of Academic Press for their cooperation. Much of the work
involved in organizing this volume was performed while I was at the
Johnson Research Foundation of the University of Pennsylvania. My
thanks and appreciation go to Dr. Britton Chance for his hospitality.
March, 9691
JOHN M. NIETSNEWOL
METHODS IN ENZYMOLOGY
DETIDE YB
Sidney .P Colowick and Nathan O. Kaplan
VANDERBILT UNIVERSITY DEPARTMENT OF CHEMISTRY
SCHOOL OF MEDICINE UNIVERStITY OF CALIFORNIA
NASHVILLE~ T~NNESSEE AT SAN DIEGO
LA JOLLA, CALIFORNIA
I. Preparation and Assay of Enzymes
II. Preparation and Assay of Enzymes
III. Preparation and Assay of Substrates
IV. for Techniques Special the Enzymologist
.V Preparation and Assay of Enzymes
.IV Preparation and Assay of Enzymes )deunitnoC(
Preparation and Assay of Substrates
Techniques Special
VII. Cumulative Subject Index
METHODS IN ENZYMOLOGY
EDITORS-IN-CHIEF
Sidney .P Colowick Nathan O. nalpaK
EMULOV VIII. Complex Carbohydrates
Edited by ELIZABETH F. NEUFELD DNA VICTOR GRUBSNIG
EMULOV IX. Carbohydrate Metabolism
Edited by WILLIS A. DOOW
EMULOV X. Oxidation and Phosphorylation
Edited by DLANOR W. KOORBATSE DNA DRANYAM E. NAMLLUP
EMULOV XI. Enzyme Structure
Edited by .C H. W. Hms
EMULOV XII. Nucleic Acids (Parts A and B)
Edited by ECNEaWAL NAMSSORG DNA KIVIE EVADLOM
EMULOV XIII. Citric Acid Cycle
Edited by J. M. NIETSNEWOL
EMULOV XIV. Lipids
Edited by J. M. NIETSNEWOL
EMULOV XV. Steroids and Terpenoids
Edited by DNOMYAR B. NOTYALC
EMULOV XVI. Fast Reactions
Edited by HTENNEK NITSUK
EMULOV XVII. Metabolism of Amino Acids and Amines (Parts A and B)
Edited by TREBREH ROBAT DNA AILEC WHITE ROBAT
EMULOV XVIII. Vitamins and Coenzymes (Parts A, B, and C)
Edited by DLANOD B. KCIMROCCM DNA LEUMEL D. THGIRW
EMULOV XIX. Proteolytic Enzymes
Edited by EDURTREG E. NNAMLREP DNA OLZSAL DNAROL
iiivx
SDOHTEM IN YGOLOMYZNE xix
VOLUME XX. Nucleic Acids and Protein Synthesis (Part )C
Edited yb KIVIE MOLDAVE AND LAWRENCE GROSSMAN
VOLUME XXI. Nucleic Acids (Part D)
Edited yb LAWRENCE NAMSSORG DNA KIVIE EVADLOM
VOLUME XXlI. Enzyme Purification and Related Techniques
Edited by WILLIAM B. YBOKAJ
VOLUME XXlII. Photosynthesis (Part A)
Edited by ANTHONY NAS PIETRO
VOLUME XXIV. Photosynthesis and Nitrogen Fixation (Part B)
Edited by ANTHONY NAS PIETRO
VOLUME XXV. Enzyme Structure (Part B)
Edited by C. H. W. HIRS DNA SERGE N. TIMASHEFF
VOLUME XXVI. Enzyme Structure (Part C)
Edited yb C. H. W. HIRS DNA SERGE N. TIMASHEFF
VOLUME XXVII. Enzyme Structure (Part D)
Edited by C. H. W. HIRS DNA SERGE N. TIMASHEFF
VOLUME XXVIII. Complex Carbohydrates (Part B)
Edited yb VICTOR GRUBSNIG
VOLUME XXIX. Nucleic Acids and Protein Synthesis (Part E)
Edited by LAWRENCE NAMSSORG DNA KIVIE EVADLOM
VOLUME XXX. Nucleic Acids and Protein Synthesis (Part F)
Edited yb KIVIE MOLDAVE DNA LAWRENCE NAMSSORG
VOLUME XXXI. Biomembranes (Part A)
Edited by SIDNEY FLEISCHER DNA LESTER REKCAP
VOLUME XXXII. Biomembranes (Part B)
Edited by SIDNEY FLEISCHER DNA LESTER REKCAP
VOLUME XXXIII. Cumulative Subject Index Volumes I-XXX
Edited yb MARTHA G. DENNIS DNA EDWARD A. DENNIS
XX METHODS IN ENZYMOLOGY
VOLUME XXXIV. Affinity Techniques (Enzyme Purification: Part B)
Edited yb WILLIAM B. YBOKAJ DNA MEIR WILCHEK
VOLUME XXXV. Lipids (Part B)
Edited by JOHN M. LOWENSTEIN
VOLUME XXXVI. Hormone Action (Part A: Steroid Hormones)
Edited by BERT W. O'MALLEY DNA JOEL G. HARDMAN
VOLUME XXXVII. Hormone Action (Part B: Peptide Hormones)
Edited by BERT W. O'MALLEY DNA JOEL G. NAMDRAH
VOLUME XXXVIII. Hormone Action (Part C: Cyclic Nucleotides)
Edited by JOEL G. HARDMAN DNA BERT W. O'MALLEY
VOLUME XXXIX. Hormone Action (Part D: Isolated Cells, Tissues, and
Organ Systems)
Edited by JOEL G. HARDMAN DNA BERT W. O'MALLEY
VOLUME XL. Hormone Action (Part E: Nuclear Structure and Function)
Edited by BERT W. O'MALLEY DNA JOEL G. HARDMAN
VOLUME XLI. Carbohydrate Metabolism (Part )B
Edited by W. A. WOOD
VOLUME XLII. Carbohydrate Metabolism (Part )C
Edited by W. A. WOOD
VOLUME XLIII. Antibiotics
Edited by JOHN H. HASH
VOLUME XLIV. Immobilized Enzymes
Edited by KLAUS HCABSOM
VOLUME XLV. Proteolytic Enzymes (Part )B
Edited by LASZLO LORAND
VOLUME XLVI. Affinity Labeling
Edited yb WILLIAM B. YBOKAJ DNA MEIR WILCHEK
VOLUME XLVII. Enzyme Structure (Part E)
Edited by C. H. W. HIRS DNA SERGE N. TIMASHEFF
METHODS IN ENZYMOLOGY xxi
VOLUME XLVIII. Enzyme Structure (Part F)
Edited yb C. H. W. HIRS DNA SERGE N. TIMASHEFF
VOLUME XLIX. Enzyme Structure (Part G)
Edited yb C. H. W. HIRS DNA SERGE N. TIMASHEFF
VOLUME L. Complex Carbohydrates (Part C)
Edited yb VICTOR GRUBSNIG
VOLUME LI. Purine and Pyrimidine Nucleotide Metabolism
Edited yb PATRICIA A. HOFFEE DNA MARY ELLEN JONES
VOLUME LII. Biomembranes (Part C: Biological Oxidations)
Edited by SIDNEY FLEISCHER DNA LESTER REKCAP
VOLUME LIII. Biomembranes (Part D: Biological Oxidations)
Edited yb SIDNEY FLEISCHER DNA LESTER REKCAP
VOLUME LIV. Biomembranes (Part E: Biological Oxidations)
Edited yb SIDNEY FLEISCHER DNA LESTER REKCAP
VOLUME LV. Biomembranes (Part F: Bioenergetics) (in preparation)
Edited by SIDNEY REI-ICSIELF DNA LESTER REKCAP
VOLUME LVI. Biomembranes (Part G: Bioenergetics) (in preparation)
Edited by SIDNEY FLEISCHER DNA LESTER REKCAP
VOLUME LVII. Bioluminescence and Chemiluminescence
Edited yb MARLENE A. DELUCA
VOLUME LVIII. Cell Culture
Edited by WILLIAM B. YBOKAJ DNA ARI H. NATSAP
VOLUME LIX. Nucleic Acids and Protein Synthesis (Part G)
Edited by KIVlE MOLDAVE DNA LAWRENCE NAMSSORG
VOLUME LX. Nucleic Acids and Protein Synthesis (Part H) (in prepa-
ration)
Edited by KIVIE MOLDAVE DNA LAWRENCE NAMSSORG