Contributors to Volume 321 Article numbers are in parentheses following the names of contributors. Affiliations listed are current. MIHCAOJ EKLHEB (3), Max Delbriick Center NOEL .S YHRAF (21, 22), Department of Inter- for Molecular Medicine, D-13122 Berlin, laH Medicine, University of Virginia Health Germany System, Charlottesville, Virginia 22908 YREME N. NWORB (17), Statistics Research NEELHTAK B. LLAH (18), Department of Bio- Laboratory, Department of Anesthesia and chemistry, Washington University School of Critical Care, Massachusetts General Hos- Medicine, Saint Louis, Missouri 01136 pital, Harvard Medical School/MIT Divi- sion of Health Sciences and Technology, YERFFEJ C. NESNAH (4), Department of Bio- Boston, Massachusetts 41120 chemistry, University of Texas Health Sci- ences Center, naS Antonio, Texas -92287 NVWLO NORYB (16), Division of Infection and 0093 Immunity, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow LEAHCIM L. NOSNHOJ (10, ,11 ,21 ,41 23, 24) 21G 8QQ, Scotland Departments of Pharmacology and Internal YNNEL M. SREHTURRAC (4), Department of Medicine, Center for Biomathematical Biochemistry, University of Texas Health Technology, National Science Foundation Sciences Center, San Antonio, Texas Science and Technology Center for Biologi- 0093-92287 cal Timing, General Clinical Research Cen- ter, University of Virginia Health System, JONATHAN B. CHAIRES (19), Department of Charlottesville, Virginia 5370-80922 Biochemistry, University of Mississippi Medical Center, Jackson, Mississippi DRAHCIR P. JUNGHANS (8), Biotherapeutics 5054-61293 Development Laboratory, Harvard Insti- NHOJ J. AIERROC (5), Department of Bio- tute of Human Genetics, Harvard Medical chemistry, University of Mississippi Medical School and Division of Hematology- Center, Jackson, Mississippi 5054-61293 Oncology, Beth Israel Deaconess Medical Center, Boston, Massachusetts 51120 LEINAD J. Cox (21, 22), Center for Behavioral Medicine Research, University of Virginia SIROB P. VEHCTAVOK (21, 22), Center for Health System, Charlottesville, Virginia Behavioral Medicine Research, University 80922 of Virginia Health System, Charlottesville, NEILUJ S. SIVAD (2), Cardiology Branch, Na- Virginia 22908 tional Heart, Lung and Blood Institute, Na- ELLEHCIM LPMAL (10, 11), Departments of tional Institutes of Health, Bethesda, Mary- Anthropology and Pediatrics, Emory Uni- land 0561-29802 versity, Atlanta, Georgia 22303 BORRIES DEMELER (3, 4), Department of Bio- chemistry, The University of Texas Health MAILLIW R. SWAL (13), Department of Bio- Sciences Center at San Antonio, San Anto- chemistry, Mount Sinai School of Medicine, nio, Texas 0093-92287 New York, New York 10029 SOILIME K. SIDAIRTIMID (7), Division of Bio- CRAM .S SIWEL (7, 8), Division of Bioen- engineering and Physical Science, ORS, gineering and Physical Science, National O D, National lnstitutes of Health, Bethesda, Institutes of Health, Bethesda, Maryland Maryland 6675-29802 6675-29802 vii viii CONTRIBUTORS TO VOLUME 321 LESLIE M. LOEW (1), Department of Physiol- VIRGIL R. SCHIRF (4), Department of Bio- ogy, Center for Biomedical Imaging Tech- chemistry, University of Texas Health Sci- nology, University of Connecticut Health ences Center, San Antonio, Texas -92287 Center, Framingham, Connecticut 03060 0093 OLE G. MOURITSEN (15), Department of CHRISTOPHER H. SCHMID (17), Biostatistics Chemistry, Technical University of Den- Research Center, Division of Clinical Care mark, DK-2800 Lyngby, Denmark Research, New England Medical Center, Boston, Massachusetts 11120 TIMOTHy G. MYERS (20), Information Tech- nology Branch, National Cancer Institute, BoRis M. SLEPCHENKO (1), Department of Physiology, Center for Biomedical Imaging National Institutes of Health, Bethesda, Technology, University of Connecticut Maryland 29802 Health Center, Framingham, Connecticut JOHN S. PHILO (6), Alliance Protein Labora- 03060 tories, Inc., Thousand Oaks, California MARTIN STRAUME (11, 12, 21, 22), Division 3282-06319 of Endocrinology and Metabolism, Depart- STEVEN M. PINCUS (9), Guilford, Connecti- ment of Internal Medicine, Center for Bio- cut 06437 mathematical Technology, National Science XIAOGANG QU (19), Department of Biochem- Foundation Center for Biological Timing, istry, University of Mississippi Medical Cen- University of Virginia Health System, Char- ter, Jackson, Mississippi 5054-61293 lottesville, Virginia 5370-80922 WILLIAM W. NAV OSDOL (20), ALZA Corpo- EDWARD L. RACHOFSKY (13), Department of ration, Mountain View, California 34049 Biochemistry, Mount Sinai School of Medi- cine, New York, New York 10029 SENNAHOJ D. VELDHUIS (14), General Clini- cal Research Center, University of Virginia OTro RISTAU (3), Max Delbriick Center for Health Sciences Center, Charlottesville, Molecular Medicine, Berlin D-13122, Virginia 2020-80922 Germany JOHN N. WEINSTEIN (20), Laboratory for Mo- MADS C. SABRA (15), Department of Chemis- lecular Pharmacology, National Cancer try, Technical University of Denmark, DK- Institute, National Institutes of Health, 2800 Lyngby, Denmark Bethesda, Maryland 29802 JAMES C. SCHAW (1), Department of Physiol- DERYCK JEREMY WILLIAMS (18), Department ogy, Center for Biomedical Imaging Tech- of Biochemistry, Washington University nology, University of Connecticut Health School of Medicine, Saint Louis, Mis- Center, Framingham, Connecticut 03060 souri 01136 Preface In the five years that have elapsed since the appearance of Part B of "Numerical Computer Methods" sdohteM( ni ,ygolomyznE Volume 240), there have been substantial enhancements in computer speed, memory size, and disk space, and decreases in computer costs. There have been parallel improvements in the numerical methodologies that can now be applied to the analysis of biological data. Procedures that previously re- quired the power of a supercomputer are now commonly run on personal computers and workstations. Computers are not oracles! But many biomedical researchers have learned to revere computers as oracles. They assume that if a computer analyzed their data the results must be correct. Data analysis algorithms make assumptions about the nature of the experimental data and the best method of analysis based on assumptions about the experimental data. They also make compromises to save computer time and space in the memory of the computer. Computer programmers also make mistakes. Consequently, computer programs can include unwarranted assumptions and can make mistakes! Biomedical researchers cannot simply insert data into a computer and accept the results as gospel. Researchers must be aware of the assumptions and methods used by their data analysis programs. They should always question the results of a computer analysis, i.e., do they have physical meaning? The purpose of these volumes is to help biomedical researchers meet these needs. It is clear that new methods of analyzing experimental data have en- hanced the type of conclusions that can be drawn and have changed the way in which experiments are conceived and done. The biochemical community must be aware of these new developments. The aim of this volume, and of Parts A and B (Volumes 210 and 240), is to inform biomedical researchers of the modern data analysis methods that have developed concomitantly with computer hardware. MICHAEL L. NOSNHOJ LUDWIG BRAND xi METHODS IN ENZYMOLOGY EMULOV I. Preparation and Assay of Enzymes detidE yb SIDNEY P. KCIWOLOC DNA NATHAN O. KAPLAN EMULOV II. Preparation and Assay of Enzymes detidE yb SIDNEY P. KCIWOLOC DNA NATHAN O. KAPLAN EMULOV Ill. Preparation and Assay of Substrates detidE by SIDNEY P. KCIWOLOC DNA NATHAN O. KAPLAN EMULOV IV. Special Techniques for the Enzymologist detidE by SIDNEY P. KCIWOLOC DNA NATHAN O. KAPLAN EMULOV V. Preparation and Assay of Enzymes detidE by SIDNEY P. KCIWOLOC DNA NATHAN O. KAPLAN EMULOV VI. Preparation and Assay of Enzymes )deunitnoC( Preparation and Assay of Substrates Special Techniques detidE yb SIDNEY P. KCIWOLOC DNA NATHAN O. KAPEAN EMULOV VII. Cumulative Subject Index detidE yb SIDNEY P. KCIWOLOC DNA NATHAN O. KAPLAN EMULOV VIII. Complex Carbohydrates detidE yb ELIZABETH F. NEUFELD DNA VICTOR GRUBSNIG EMULOV IX. Carbohydrate Metabolism detidE yb WILLIS A. WOOD EMULOV X. Oxidation and Phosphorylation detidE by RONALD W. KOORBATSE DNA MAYNARD E. NAMLLUP EMULOV XI. Enzyme Structure detidE yb C. H. W. HIRS EMULOV XII. Nucleic Acids (Parts A and B) detidE yb ECNERWAL NAMSSORG DNA KIVIE EVADLOM EMULOV XIII. Citric Acid Cycle detidE yb J. M. NIETSNEWOL EMULOV XIV. Lipids detidE yb J. M. NIETSNEWOL EMULOV XV. Steroids and Terpenoids detidE by DNOMYAR B. NOTYALC EMULOV XVI. Fast Reactions detidE yb NITSUK HTENNEK xi xii METHODS IN ENZYMOLOGY EMULOV XVII. Metabolism of Amino Acids and Amines (Parts A and B) Edited by TREBREH ROBAT DNA CELIA ETIHW 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 EMULOV XX. Nucleic Acids and Protein Synthesis (Part )C Edited by KIVIE EVADLOM DNA ECNERWAL NAMSSORG EMULOV XX|. Nucleic Acids (Part D) Edited by ECNERWAL NAMSSORG DNA KIVIE EVADLOM EMULOV XXII. Enzyme Purification and Related Techniques Edited by MAILLIW B. YBOKAJ EMULOV XXIII. Photosynthesis (Part A) Edited by YNOHTNA NAS ORTEIP EMULOV XXIV. Photosynthesis and Nitrogen Fixation (Part B) Edited by YNOHTNA NAS ORTEIP EMULOV XXV. Enzyme Structure (Part B) Edited by .C H. W. HIRS DNA EGRES N. FFEHSAMIT EMULOV XXVI. Enzyme Structure (Part C) Edited by .C H. W. HIRS DNA EGRES N. FFEHSAMIT EMULOV XXVII. Enzyme Structure (Part D) Edited by C. H. W. HIRS DNA EGRES N. FFEHSAMIT EMULOV XXVIII. Complex Carbohydrates (Part B) Edited by ROTCIV GRUBSNIG EMULOV XXIX. Nucleic Acids and Protein Synthesis (Part E) Edited by ECNERWAL NAMSSORG DNA KIVIE EVADLOM EMULOV XXX. Nucleic Acids and Protein Synthesis (Part F) Edited by KIVIE EVADLOM DNA ECNERWAL NAMSSORG EMULOV XXXI. Biomembranes (Part A) Edited by YENDIS REHCSIELF DNA RETSEL REKCAP EMULOV XXXII. Biomembranes (Part B) Edited by YENDIS REHCSIELF DNA RETSEL REKCAP EMULOV XXXIII. Cumulative Subject Index Volumes I-XXX Edited by AHTRAM G. SINNED DNA DRAWDE A. SINNED EMULOV XXXIV. Affinity Techniques (Enzyme Purification: Part B) Edited by MAILLIW B. YBOKAJ DNA MEIR KEHCLIW EMULOV XXXV. Lipids (Part B) Edited by NHOJ M. NIETSNEWOL METHODS NI ENZYMOLOGY Xlll .,° EMULOV XXXVI. Hormone Action (Part A: Steroid Hormones) Edited by BERT W. O'MALLEY DNA JOEL G. NAMDRAH EMULOV XXXVII. Hormone Action (Part B: Peptide Hormones) Edited by BERT W. O'MALLEY DNA JOEL G. NAMDRAH EMULOV XXXVIII. Hormone Action (Part C: Cyclic Nucleotides) Edited by JOEL G. HARDMAN DNA BERT W. O'MALLEY EMULOV XXXIX. Hormone Action (Part D: Isolated Cells, Tissues, and Organ Systems) Edited by JOEL G. HARDMAN DNA BERT W. O'MALLEY EMULOV XL. Hormone Action (Part E: Nuclear Structure and Function) Edited by BERT W. O'MALLEY DNA JOEL G. HARDMAN EMULOV XLI. Carbohydrate Metabolism (Part B) Edited by W. A. DOOW EMULOV XLII. Carbohydrate Metabolism (Part C) Edited by W. A. DOOW EMULOV XLIII. Antibiotics Edited by JoHN H. HASH EMULOV XLIV. Immobilized Enzymes Edited by SUALK HCABSOM EMULOV XLV. Proteolytic Enzymes (Part B) Edited by OLZSAL DNAROL EMULOV XLVI. Affinity Labeling Edited by WILLIAM B. JAKOBY DNA MEIR WILCHEK EMULOV XLVII. Enzyme Structure (Part E) Edited by C. H. W. HIRS DNA EGRES N. FFEHSAMIT EMULOV XLVIII. Enzyme Structure (Part F) Edited by C. H. W. HIRS DNA EGRES N. FFEHSAMIT EMULOV XLIX. Enzyme Structure (Part G) Edited by C. H. W. HIRS DNA EGRES N. FFEHSAMIT EMULOV L. Complex Carbohydrates (Part C) Edited by VICrOR 6RUBSNIG EMULOV LI. Purine and Pyrimidine Nucleotide Metabolism Edited by PATRICIA A. HOFFEE DNA MARY ELLEN JONES EMULOV LII. Biomembranes (Part C: Biological Oxidations) Edited by SIDNEY FLEISCHER DNA LESTER PACKER EMULOV LIII. Biomembranes (Part D: Biological Oxidations) Edited by SIDNEY FLEISCHER DNA LESTER PACKER EMULOV LIV. Biomembranes (Part E: Biological Oxidations) Edited by YENDIS FLEISCHER DNA LESTER PACKER xiv METHODS IN ENZYMOLOGY EMULOV LV. Biomembranes (Part F: Bioenergetics) Edited yb SIDNEY FLEISCHER AND LESTER PACKER EMULOV LVI. Biomembranes (Part G: Bioenergetics) Edited by SIDNEY FLEISCHER AND LESTER PACKER EMULOV LVII. Bioluminescence and Chemiluminescence Edited by MARLENE A. DELUCA EMULOV LVIII. Cell Culture Edited by WILLIAM B. YBOKAJ AND IRA PASTAN EMULOV LIX. Nucleic Acids and Protein Synthesis (Part G) Edited yb KIVIE MOLDAVE AND LAWRENCE GROSSMAN EMULOV LX. Nucleic Acids and Protein Synthesis (Part H) Edited by KIVIE MOLDAVE AND LAWRENCE GROSSMAN EMULOV .16 Enzyme Structure (Part H) Edited by C. H. W. HIRS DNA EGRES N. FEEHSAMIT EMULOV .26 Vitamins and Coenzymes (Part D) Edited yb DONALD B. McCORMICK AND LEMUEL D. WRIGHT EMULOV .36 Enzyme Kinetics and Mechanism (Part A: Initial Rate and Inhibitor Methods) Edited by DANIEL L. PURICH EMULOV .46 Enzyme Kinetics and Mechanism (Part B: Isotopic Probes and Com- plex Enzyme Systems) Edited by DANIEL L. PURICH EMULOV .56 Nucleic Acids (Part I) Edited yb LAWRENCE GROSSMAN AND EIVIK MOLDAVE EMULOV .66 Vitamins and Coenzymes (Part E) Edited by DONALD B. McCORMICK AND LEMUEL D. WRIGHT EMULOV .76 Vitamins and Coenzymes (Part F) Edited yb DONALD B. McCORMICK AND LEMUEL D. WRIGHT VOLUME 68. Recombinant DNA Edited yb YAR Wu EMULOV .96 Photosynthesis and Nitrogen Fixation (Part C) Edited by ANTHONY SAN PIETRO EMULOV .07 Immunochemical Techniques (Part A) Edited yb HELEN VAN VUNAKIS AND NHOJ J. LANGONE EMULOV .17 Lipids (Part C) Edited by NHOJ M. NIETSNEWOL EMULOV .27 Lipids (Part D) Edited by NHOJ M. NIETSNEWOL METHODS IN ENZYMOLOGY VX EMULOV .37 Immunochemical Techniques (Part B) Edited by JOHN J. ENOGNAL DNA HELEN NAV SIKANUV EMULOV .47 Immunochemical Techniques (Part C) Edited by JOHN J. LANGONE AND HELEN VAN VUNAKIS EMULOV .57 Cumulative Subject Index Volumes XXXI, XXXII, XXXIV-LX Edited by DRAWDE A. DENNIS AND MARTHA G. DENNIS EMULOV .67 Hemoglobins Edited by ERALDO ANTONINI, LUIGI ROSsI-BERNARDI, AND EMILIA CH1ANCONE EMULOV .77 Detoxication and Drug Metabolism Edited by WILLIAM B. JAKOBY EMULOV .87 Interferons (Part A) Edited by YENDIS AKTSEP EMULOV .97 Interferons (Part B) Edited by YENDIS AKTSEP EMULOV .08 Proteolytic Enzymes (Part C) Edited by OLZSAL DNAROL EMULOV .18 Biomembranes (Part H: Visual Pigments and Purple Mem- branes, I) Edited by RETSEL REKCAP EMULOV .28 Structural and Contractile Proteins (Part A: Extracellular Matrix) Edited by LEON W. MAHGNINNUC DNA DIXIE W. NESKIREDERF EMULOV .38 Complex Carbohydrates (Part D) Edited by ROTCIV GRUBSNIG EMULOV .48 Immunochemical Techniques (Part D: Selected Immunoassays) Edited by JOHN J. LANGONE AND HELEN VAN VUNAKIS EMULOV .58 Structural and Contractile Proteins (Part B: The Contractile Appara- tus and the Cytoskeleton) Edited by DIXIE W. FREDERIKSEN AND LEON W. CUNNINGHAM EMULOV .68 Prostaglandins and Arachidonate Metabolites Edited by WILLIAM E. M. LANDS AND WILLIAM L. SMITH EMULOV .78 Enzyme Kinetics and Mechanism (Part C: Intermediates, Stereo- chemistry, and Rate Studies) Edited by DANIEL L. PURICH EMULOV .88 Biomembranes (Part I: Visual Pigments and Purple Mem- branes, II) Edited by RETSEL REKCAP EMULOV .98 Carbohydrate Metabolism (Part D) Edited by WILLIS A. WooD EMULOV .09 Carbohydrate Metabolism (Part E) Edited by WILLIS A. WOOD xvi METHODS NI ENZYMOLOGY EMULOV 91. Enzyme Structure (Part I) Edited yb C. H. W. HIRS DNA SERGE N. FFEHSAMIT EMULOV 92. Immunochemical Techniques (Part E: Monoclonal Antibodies and General Immunoassay Methods) Edited by JOHN J. LANGONE DNA HELEN VAN SIKANUV EMULOV 93. Immunochemical Techniques (Part F: Conventional Antibodies, Fc Receptors, and Cytotoxicity) Edited by JOHN J. LANGONE DNA HELEN VAN SIKANUV EMULOV 94. Polyamines Edited by HERBERT TABOR DNA CELIA WHITE TABOR EMULOV 95. Cumulative Subject Index Volumes 61-74, 76-80 Edited by EDWARD A. DENNIS DNA MARTHA G. SINNED EMULOV 96. Biomembranes Part J: Membrane Biogenesis: Assembly and Tar- geting (General Methods; Eukaryotes) Edited yb YENDIS REHCSIELF DNA BECCA REHCSIELF EMULOV 97. Biomembranes Part K: Membrane Biogenesis: Assembly and Tar- geting (Prokaryotes, Mitochondria, and Chloroplasts) Edited by YENDIS REHCSIELF DNA BECCA REHCSIELF EMULOV 98. Biomembranes (Part L: Membrane Biogenesis: Processing and Re- cycling) Edited by YENDIS REHCSIELF DNA BECCA REHCSIELF EMULOV 99. Hormone Action (Part F: Protein Kinases) Edited by JACKIE O. CORBIN DNA JOEL G. NAMDRAH EMULOV 100. Recombinant DNA (Part B) Edited by RAY Wu, ECNERWAL ,NAMSSORG DNA KIVIE EVADLOM EMULOV 101. Recombinant DNA (Part C) Edited by RAY Wu, ECNERWAL ,NAMSSORG DNA KIVIE EVADLOM EMULOV 102. Hormone Action (Part G: Calmodulin and Calcium-Binding Pro- teins) Edited yb ANTHONY R. MEANS DNA BERT W. O'MALLEY EMULOV 103. Hormone Action (Part H: Neuroendocrine Peptides) Edited by P. MICHAEL NNOC EMULOV 104. Enzyme Purification and Related Techniques (Part )C Edited by WILLIAM B. YBOKAJ EMULOV 105. Oxygen Radicals in Biological Systems Edited by LESTER REKCAP EMULOV 106. Posttranslational Modifications (Part )A Edited by FINN WOLD DNA KIVIE EVADLOM EMULOV 107. Posttranslational Modifications (Part )B Edited by FINN WOLD DNA KIVIE EVADLOM SDOHTEM NI YGOLOMYZNE xvii EMULOV 108. Immunochemical Techniques (Part G: Separation and Characteriza- tion of Lymphoid Cells) Edited by GIOVANNI DI SABATO, JOHN J. LANGONE, DNA HELEN VAN SIKANUV EMULOV 109. Hormone Action (Part I: Peptide Hormones) Edited by ZTUL REMUABNRIB DNA TREB W. YELLAM'O EMULOV 110. Steroids and Isoprenoids (Part )A Edited by NHOJ H. WAL DNA SNAH .C GNILL1R EMULOV 111. Steroids and Isoprenoids (Part B) Edited by NHOJ H. WAL DNA SNAH .C GNILLIR EMULOV 112. Drug and Enzyme Targeting (Part A) Edited by HTENNEK J. REDDIW DNA HPLAR NEERG EMULOV 113. Glutamate, Glutamine, Glutathione, and Related Compounds Edited by NOTLA RETSIEM EMULOV 114. Diffraction Methods for Biological Macromolecules (Part A) Edited by DLORAH W. ,FFOKCYW .C H. W. HIRS, DNA EGRES N. FFEHSAMIT EMULOV 115. Diffraction Methods for Biological Macromolecules (Part B) Edited by DLORAH W. ,FEOKCYW .C H. W. HIRS, DNA EGRES N. FFEHSAMIT EMULOV 116. Immunochemical Techniques (Part H: Effectors and Mediators of Lymphoid Cell Functions) Edited by GIOVANNI DI ,OTABAS JOHN J. LANGONE, DNA HELEN VAN SIKANUV EMULOV 117. Enzyme Structure (Part J) Edited by .C H. W. HIRS DNA EGRES N. FFEHSAMIT EMULOV 118. Plant Molecular Biology Edited by RUHTRA HCABSSIEW DNA TREBREH HCABSSIEW EMULOV 119. Interferons (Part C) Edited by YENDIS AKTSEP EMULOV 120. Cumulative Subject Index Volumes 81-94, 96-101 EMULOV 121. Immunochemical Techniques (Part I: Hybridoma Technology and Monoclonal Antibodies) Edited by NHOJ J. ENOGNAL DNA NELEH NAV SIKANUV EMULOV 122. Vitamins and Coenzymes (Part G) Edited by KNARF LITYHC DNA DLANOD B. KCIMRoCcM EMULOV 123. Vitamins and Coenzymes (Part )H Edited by KNARF LITYHC DNA DLANOD B. KCIMRoCcM EMULOV 124. Hormone Action (Part J: Neuroendocrine Peptides) Edited by P. LEAHCIM NNOC