Contributors to Volume 226 elcitrA srebmun era ni sesehtnerap gniwollof the seman of .srotubirtnoc snoitailiffA listed era .tnerruc DAVID S. AULD (23), Center for Biochem- TERESA B. FREEDMAN (13, 19), Department ical and Biophysical Sciences and Medi- of Chemistry, Syracuse University, 410-1 cine, Harvard Medical School, Boston, Center for Science and Technology, Syra- Massachusetts 51120 cuse, New York 44231 JANINA C. AUSTIN (15), Department of SIMON J. GEORGE (9), Department of Ap- Chemistry, Princeton University, Prince- plied Science, University of California, ton, New Jersey 44580 Davis, Davis, California 61659 BRUCE A. AVERILL (2), Department of ROBERT A. GOLDBECK (7), Department of Chemistry, University of Virginia, Chemistry and Biochemistry, University Charlottesville, Virginia 10922 of California, Santa Cruz, Santa Cruz, GERALD T. BABCOCK (17), The LASER Lab- California 46059 oratory, and Department of Chemistry, WILLIAM DEW. HORROCKS, JR. (21), De- Michigan State University, East Lansing, partment of Chemistry, The Pennsylvania Michigan 42884 State University, University Park, Penn- JAcQUELINE K. BARTON (25), Division of sylvania 20861 Chemistry and Chemical Engineering, DAVID S. KLIGER (7), Department of Chem- ,03-461 California Institute of Technol- istry and Biochemistry, University of -laC ogy, Pasadena, California 52119 ifornia, Santa Cruz, Santa Cruz, Califor- MARK R. CHANCE (5, 6), Department of ain 46059 Physiology and Biophysics, Albert Ein- NENAD M. KOSTI~ (24), Department of stein College of Medicine, Bronx, New Chemistry, Iowa State University, Ames, York 16401 Iowa 11005 MYLES R. CHEESMAN (9), Center for Metal- Louis B. LACROIX (1), Department of loprotein Spectroscopy and Biology, Chemistry, Stanford University, Stan- School of Chemical Sciences, University ford, California 50349 of East Anglia, Norwich NR4 7T J, United JAMES A. LARRABEE (12), Department of Kingdom Chemistry and Biochemistry, Science EEFEI CHEN (6), Department of Chemistry, Center, Middlebury College, Middlebury, University of California, Santa Cruz, Vermont 35750 Santa Cruz, California 46059 THOMAS M. LOEHR (18), Department of SUNHEE CHOI (12), Department of Chemis- Chemical and Biological Sciences, Ore- yrt and Biochemistry, Science Center, gon Graduate Institute of Science and Middlebury College, Middlebury, Ver- Technology, Beaverton, Oregon 60079 mont 35750 MICHAEL D. LOWERY (1), Department of ROMAN S. CZERNUSZEWICZ (11), Depart- Chemistry, Stanford University, Stan.. ment of Chemistry, University of Hous- ford, California 50349 ton, Houston, Texas 40277 WOLFGANG MARET (3, 26), Center for Bio- ROBERT C. DUNN (8), Pacific Northwest chemical and Biophysical Sciences and Laboratories, Battelle, Richland, Wash- Medicine, Harvard Medical School, Bos- ington 25399 ton, Massachusetts 511'20 vii Vlll CONTRIBUTORS TO VOLUME 226 CATHERINE J. MURPHY (25), Division of GIULIETTA SMULEVICH (16), Laboratorio di Chemistry and Chemical Engineering, Spettroscopia Molecolare, Dipartimento ,03-461 California Institute of Technol- di Chimica, Universita di Firenze, 12105 ogy, Pasadena, California 52119 Florence, Italy EDWARD I. SOLOMON (1), Department of LAURENCE A. NAFIE 03, 19), Department Chemistry, Stanford University, Stan- of Chemistry, Syracuse University, 410-1 ford, California 50349 Center for Science and Technology, Syra- cuse, New York 44231 THOMAS G. SPIRO (15, 16), Department of Chemistry, Princeton University, Prince- KAZUO NAKAMOTO (11), Department of ton, New Jersey 44580 Chemistry, Marquette University, Mil- ANDREW J. THOMSON (9), Center for Metal- waukee, Wisconsin 33235 loprotein Spectroscopy and Biology, THOMAS V. O'HALLORAN (4), Department School of Chemical Sciences, University of Chemistry, Northwestern University, of East Anglia, Norwich NR 4 7TJ, Evanston, Illinois 80206 United Kingdom LISA M. UTSCHIG (4), Department of Chem- FREDERICK S. RICHARDSON (22), Depart- istry, Northwestern University, Evanston, ment of Chemistry, University of Vir- Illinois 80206 ginia, Charlottesville, Virginia 10922 BERT L. VALLEE (3), Center for Biochem- JAMES P. RIEHL (22), Department of Chem- ical and Biophysical Sciences and Medi- istry, University of Missouri, St. Louis, cine, Harvard Medical School, Boston, St. Louis, Missouri 12136 Massachusetts 51120 ALISON RODGER (10), Physical Chemistry HAROLD E. VAN WART (14), Institute of Biochemistry and Cell Biology, Syntex Laboratory, Oxford University, Oxford Discovery Research, Paid Alto, Califor- 1(5.0 3QZ, England nia 30349 KENTON R. ROGERS (15), Department of SONITNATSNOC VAROTSIS (17), ehT LASER Chemistry, Princeton University, Prince- Laboratory, and Department of Chemis- ton, New Jersey 44580 try, Michigan State University, East DAVID E. ROOT (1), Department of Chemis- Lansing, Michigan 42884 try, Stanford University, Stanford. Cali- JOHN B. VINCENT (2), Department of fornia 50349 Chemistry, University of Alabama, Tus- caloosa, Alabama 78453 JOANN SANDERS-LOEHR (18), Department YANG WANG (14), George R. Harrison, of Chemical and Biological Sciences, -rO Spectroscopy Laboratory, Massachusetts egon Graduate Institute of Science and Institute of Technology, Cambridge, Technology, Beaverton, Oregon 60079 Massachusetts 93120 JOHN D. SIMON (8), Department of Chemis- JEFFREY G. WRIGHT (4), Department of try, University of California, San Diego, Chemistry, Northwestern University, La Jolla, California 39029 Evanston, Illinois 80206 W. E. SMITH (20), Department of Pure and XIAOLIANG XIE (8), Pacific Northwest Lab- Applied Chemistry, University of Strath- oratories, Battelle, Richland, Washington clyde, Glasgow 1G IXL, Scotland 25399 Preface The scope of metallobiochemistry has greatly expanded in recent years as ever more powerful techniques have been brought to bear on the constituent elements that define and characterize the subject. Metallobio- chemistry, Part A, Volume 851 of Methods in Enzymology, focused on progress in those areas in which early on there were major impediments to growth and development. Without the tools to measure metals with suffi- cient accuracy, precision, and sensitivity this scientific discipline could not have existed. In addition, unless it were possible to distinguish be- tween the metals present in a biological sample that belonged there (be- cause nature intended them to be) from those that merely appeared by accident (adventitious contamination), such metal analyses would have been meaningless. Technology overcame these hurdles, and Part A de- tailed the state of trace element analysis and the various approaches employed by the metallobiochemist to avoid artifacts and achieve the inorganic equivalent of microbiological sterility. It is a summary of the critical methods that have helped place the subject on a solid foundation. Metallobiochemistry, Part B, Volume 205 of this series, is devoted to metallothionein and related molecules. It is unusual for a Methods in Enzymology volume to feature a single molecule, but the surge of interest in metallothionein and its structural and possibly functional relationship to DNA-binding proteins suggested that such a volume would be timely and useful. It also seemed appropriate to stress that not all metal-contain- ing biological molecules are metalloenzymes or electron-transport pro- teins. Parts C and D, Volumes 226 and 227, respectively, return to the theme of methods that have contributed to and are emerging as important factors in the advancement of the field. These methods embody concepts that had their origin about the time of the almost forgotten Sumner-Willstfitter controversy of the 1920s. Proteins, it was claimed, could hardly serve as specific biological catalysts if they were little more than nondescript col- loids. Metal ions would prove to be the real actors on the enzymatic stage. Authority prevailed until the crystallization of urease seemingly dis- patched the metal dogma to oblivion. Despite the extended protests of interest-vested diehards, protein chemistry became inextricably associ- ated with enzymology and metals fell out of fashion. (Ironically, urease turned out to be a nickel enzyme.) Biochemists who had witnessed this metal-induced brouhaha were understandably reluctant to resurrect the idea that metals might have something to do with biological catalysis. Anyone wishing to make the case would have to have persistence along with persuasive and unassail- ix X ECAFERP able analytical data. Only through scrupulous attention to detail was it possible in those neonatal days of metallobiochemistry to gain the accep- tance that allowed the field to grow and flourish. Despite its shortcomings the metal-cum-colloid view of catalysis did have one rather appealing feature: the metal would have unique proper- ties among all the atoms of the protein and perhaps these could be ex- ploited to gain important information. The metal could serve as a beacon to guide the investigator searching for an active site. It could also be a signal, either of the detailed steps of catalysis or of any other biological function with which the metal might be associated. Emission spectros- copy proved the significance of a metal-derived signal in principle, but was rather inconsiderate of the protein. Hence, attention shifted along with wavelength to absorption spectroscopy whereby it became possible to view the functional heart of a metalloenzyme directly. This window on the world of metallobiochemistry revealed unprecedented spectral fea- tures clearly indicative of an unusual coordination environment and likely characteristic of a catalytic site. Not all metals lend themselves to absorption spectroscopic investiga- tion. Zinc, one of nature's most recurrent participants, is notoriously shy in this regard. Other metals are more expressive and revealing when viewed by alternative techniques. In these two volumes (226 and 227) we have assembled a broad representation of the physical and spectroscopic methods now available that can be useful for examining metals in biologi- cal systems and for probing their environments in metalloproteins and metalloenzymes. These approaches, while by no means all-inclusive, ex- emplify the wide variety of tools and the level of sophistication currently being applied to extract both the nuances and the general principles of metallobiochemistry pertinent to these systems. We are extremely grateful to our contributors for their willingness to participate in this endeavor. They have made a concerted effort to de- scribe techniques in ways that would be most beneficial to the reader. The chapters differ from the more typical ones in this series in that they identify principles underlying a particular method, the kinds of questions that can be addressed, and the ways to interpret results. Step-by-step instructions were not practical in most cases, and generally the objective has been to provide a sense of what can be accomplished. It required more description than anticipated for most of the topics, and this necessi- tated two volumes instead of one. We appreciate the understanding of our colleagues at Academic Press and we thank them again as well as all the contributors for making this such a pleasant experience. JAMES F. RIORDAN BERT L. VALLEE METHODS IN ENZYMOLOGY EMULOV I. Preparation and Assay of Enzymes Edited yb SIDNEY P. KCIWOLOC DNA NAHTAN O. NALPAK EMULOV II. Preparation and Assay of Enzymes Edited yb SIDNEY P. KCIWOLOC DNA NAHTAN O. NALPAK EMULOV III. Preparation and Assay of Substrates Edited yb SIDNEY P. KCIWOLOC DNA NAHTAN O. NALPAK EMULOV IV. Special Techniques for the Enzymologist Edited yb YENDIS P. KCIWOLOC DNA NAHTAN O. NALPAK EMULOV V. Preparation and Assay of Enzymes Edited yb SIDNEY P. KCIWOLOC DNA NAHTAN O. NALPAK EMULOV VI. Preparation and Assay of Enzymes )deunitnoC( Preparation and Assay of Substrates Special Techniques Edited yb SIDNEY P. KCIWOLOC DNA NAHTAN O. NALPAK EMULOV VII. Cumulative Subject Index Edited yb SIDNEY P. KCIWOLOC DNA NAHTAN O. NALPAK EMULOV VIII. Complex Carbohydrates Edited yb HTEBAZILE F. NEUFELD DNA ROTCIV GRUBSNIG EMULOV IX. Carbohydrate Metabolism Edited yb WILLIS A. DOOW EMULOV X. Oxidation and Phosphorylation Edited yb DLANOR W. KOORBATSE DNA DRANYAM E. NAMLLUP EMULOV XI. Enzyme Structure Edited yb C. H. W. HIRS EMULOV XII. Nucleic Acids (Parts A and B) Edited yb ECNERWAL NAMSSORG DNA KIVIE EVADLOM EMULOV XIII. Citric Acid Cycle Edited yb J. M. NIETSNEWOL EMULOV XIV. Lipids Edited yb J. M. NIETSNEWOL EMULOV XV. Steroids and Terpenoids Edited yb DNOMYAR B. NOTYALC EMULOV XVI. Fast Reactions Edited yb HTENNEK NITSUK ix xii METHODS IN ENZYMOLOGY EMULOV XVII. Metabolism of Amino Acids and Amines (Parts A and B) Edited by TREBREH ROBAT DNA CELIA WHITE ROBAT EMULOV XVIII. Vitamins and Coenzymes (Parts A, B, and C) Edited yb DLANOD B. McCoRMICK DNA LEUMEL D. THGIRW EMULOV XIX. Proteolytic Enzymes Edited yb EDURTREG E. NNAMLREP DNA OLZSAL DNAROL EMULOV XX. Nucleic Acids and Protein Synthesis (Part )C Edited yb KIVIE EVADLOM DNA ECNERWAL NAMSSORG EMULOV XXI. Nucleic Acids (Part D) Edited yb ECNERWAL NAMSSORG DNA KIVIE EVADLOM EMULOV XXII. Enzyme Purification and Related Techniques Edited yb WILLtAM B. YBOKAJ EMULOV XXIII. Photosynthesis (Part A) Edited yb YNOHTNA NAS ORTEIP EMULOV XXIV. Photosynthesis and Nitrogen Fixation (Part B) Edited yb YNOHTNA NAS ORTEIP EMULOV XXV. Enzyme Structure (Part B) Edited yb C. H. W. HIRS DNA EGRES N. FFEHSAMIT EMULOV XXVI. Enzyme Structure (Part C) Edited yb 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 yb ROTCIV GRUBSNIG EMULOV XXIX. Nucleic Acids and Protein Synthesis (Part E) Edited yb ECNERWAL NAMSSORG DNA KIVIE EVADLOM EMULOV XXX. Nucleic Acids and Protein Synthesis (Part F) Edited yb KIVIE EVADLOM DNA ECNERWAL NAMSSORG EMULOV XXXI. Biomembranes (Part A) Edited yb YENDIS REHCSIELF DNA RETSEL REKCAP EMULOV XXXII. Biomembranes (Part B) Edited yb SIDNEY REHCSIELF DNA RETSEL REKCAP EMULOV XXXIII. Cumulative Subject Index Volumes I-XXX Edited yb AHTRAM G. DENNIS DNA DRAWDE a. SINNED EMULOV XXXIV. Affinity Techniques (Enzyme Purification: Part B) Edited yb WILLIAM B. YBOKAJ DNA MEIR KEHCLIW EMULOV XXXV. Lipids (Part B) Edited yb JOHN M. NIETSNEWOL METHODS IN ENZYMOLOGY xiii EMULOV XXXVI. Hormone Action (Part A: Steroid Hormones) Edited yb BERT W. O'MALLEY AND JOEL G. HARDMAN EMULOV XXXVII. Hormone Action (Part B: Peptide Hormones) Edited yb BERT W. O'MALLEY DNA JOEL G. NAMDRAH EMULOV XXXVIII. Hormone Action (Part C: Cyclic Nucleotides) Edited yb JOEL G. HARDMAN AND BERT W. O'MALLEY EMULOV XXXIX. Hormone Action (Part D: Isolated Cells, Tissues, and Organ Systems) Edited by JOEL G. HARDMAN AND BERT W. O'MALLEY EMULOV XL. Hormone Action (Part E: Nuclear Structure and Function) Edited by BERT W. O'MALLEY DNA JOEL G. NAMDRAH 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 yb KLAUS HCABSOM EMULOV XLV. Proteolytic Enzymes (Part B) Edited yb OLZSAL DNAROL EMULOV XLVI. Affinity Labeling Edited yb WILLIAM B. YBOKAJ DNA MEIR KEHCLIW EMULOV XLVII. Enzyme Structure (Part E) Edited yb C. H. W. HIRS DNA EGRES N. EEEHSAMIT EMULOV XLVIII. Enzyme Structure (Part F) Edited by C. H. W. HtRS DNA EGRES N. FFEHSAMIT EMULOV XLIX. Enzyme Structure (Part G) Edited yb C. H. W. HIRS DNA EGRES N. FFEHSAMIT EMULOV L. Complex Carbohydrates (Part )C Edited by ROTCIV GRUBSNIG EMULOV LI. Purine and Pyrimidine Nucleotide Metabolism Edited yb PATRICIA A. HOFFEE AND MARY ELLEN JONES EMULOV LII. Biomembranes (Part C: Biological Oxidations) Edited yb SIDNEY FLEISCHER AND LESTER PACKER EMULOV LIII. Biomembranes (Part D: Biological Oxidations) Edited yb SIDNEY FLEISCHER AND LESTER PACKER EMULOV LIV. Biomembranes (Part E: Biological Oxidations) Edited yb SIDNEY FLEISCHER AND LESTER PACKER xiv SDOHTEM IN YGOLOMYZNE EMULOV LV. Biomembranes (Part F: Bioenergetics) Edited yb SIDNEY REHCSIELF DNA RETSEL REKCAP EMULOV LVI. Biomembranes (Part G: Bioenergetics) Edited yb SIDNEY REHCSIELF DNA RETSEL REKCAP EMULOV LVII. Bioluminescence and Chemiluminescence Edited yb ENELRAM A. DELUCA EMULOV LVIII. Cell Culture Edited yb WILLIAM B. YBOKAJ DNA ARI NATSAP EMULOV LIX. Nucleic Acids and Protein Synthesis (Part G) Edited yb KIVIE MOLDAVE DNA LAWRENCE NAMSSORG EMULOV LX. Nucleic Acids and Protein Synthesis (Part H) Edited yb KIVIE MOEDAVE DNA LAWRENCE NAMSSORG EMULOV 61. Enzyme Structure (Part H) Edited yb C. H. W. HIRS DNA EGRES N. EEEHSAMIT EMULOV .26 Vitamins and Coenzymes (Part D) Edited yb DONALD B. McCoRMICK DNA LEMUEL D. WRIGHT EMULOV 63. Enzyme Kinetics and Mechanism (Part A: Initial Rate and Inhibi- tor Methods) Edited by DANIEL L. PURICH EMULOV .46 Enzyme Kinetics and Mechanism (Part B: Isotopic Probes and Complex Enzyme Systems) Edited yb DANIEL L. PURICH EMULOV .56 Nucleic Acids (Part I) Edited by LAWRENCE NAMSSORG DNA KIVIE MOLDAVE EMULOV .66 Vitamins and Coenzymes (Part E) Edited yb DONALD B. McCoRMICK DNA LEMUEL D. WRIGHT EMULOV .76 Vitamins and Coenzymes (Part F) Edited yb DLANOD B. KCIMROCCM DNA LEUMEL D. THGIRW VOLUME 68. Recombinant DNA Edited yb YAR Wu EMULOV 69. Photosynthesis and Nitrogen Fixation (Part C) Edited yb YNOHTNA NAS ORTEIP EMULOV 70. Immunochemical Techniques (Part A) Edited yb HELEN VAN VUNAKIS DNA NHOJ J. LANGONE EMULOV 71. Lipids (Part C) Edited yb JOHN M. NIETSNEWOL EMULOV .27 Lipids (Part D) Edited yb JOHN M. NIETSNEWOL EMULOV 73. Immunochemical Techniques (Part )B Edited yb JOHN J. LANGONE DNA HELEN VAN VUNAKIS SDOHTEM IN YGOLOMYZNE VX EMULOV 74. Immunochemical Techniques (Part C) Edited by JOHN J. LANGONE DNA HELEN NAV SIKANUV EMULOV 75. Cumulative Subject Index Volumes XXXI, XXXII, XXXIV-LX Edited yb EDWARD A. DENNIS DNA MARTHA G. DENNIS EMULOV 76. Hemoglobins Edited by ERALDO ANTONINI, LUIGI ROSSI-BERNARDI, DNA EMILIA CHIAN- ENOC EMULOV 77. Detoxication and Drug Metabolism Edited yb WILLIAM B. JAKOaY EMULOV 78. Interferons (Part A) Edited yb SIDNEY AKTSEP EMULOV 79. Interferons (Part B) Edited yb SIDNEY AKTSEP EMULOV 80. Proteolytic Enzymes (Part C) Edited yb LASZLO DNAROL EMULOV .18 Biomembranes (Part H: Visual Pigments and Purple Mem- branes, I) Edited by LESTER REKCAP EMULOV 82. Structural and Contractile Proteins (Part A: ExtraceUular Matrix) Edited by LEON W. CUNNINGHAM DNA DIXIE W. FREDERIKSEN EMULOV 83. Complex Carbohydrates (Part D) Edited yb VICTOR GRUBSNIG EMULOV 84. Immunochemical Techniques (Part D: Selected Immunoassays) Edited yb JOHN J. ENOGNAL DNA HELEN NAV SIKANUV EMULOV 85. Structural and Contractile Proteins (Part B: The Contractile Appa- ratus and the Cytoskeleton) Edited yb DIXIE W. FREDERIKSEN DNA LEON W. CUNNINGHAM EMULOV 86. Prostaglandins and Arachidonate Metabolites Edited yb WILLIAM E. M. LANDS DNA WILLIAM L. SMITH EMULOV 87. 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 LESTER REKCAP EMULOV 89. Carbohydrate Metabolism (Part D) Edited by WILLIS A. WOOD EMULOV 90. Carbohydrate Metabolism (Part E) Edited yb WILLIS A. WOOD EMULOV 91. Enzyme Structure (Part I) Edited by C. H. W. HIRS DNA SERGE N. EEEHSAMIT xvi SDOHTEM NI YGOLOMYZNE EMULOV .29 Immunochemical Techniques (Part E: Monoclonal Antibodies and General Immunoassay Methods) Edited yb JOHN J. ENOCNAL DNA HELEN NAV SIKANUV EMULOV .39 Immunochemical Techniques (Part F: Conventional Antibodies, Fc Receptors, and Cytotoxicity) Edited by JOHN J. ENOGNAL DNA HELEN NAV SIKANUV EMULOV 94. Polyamines Edited yb TREBREH ROBAT DNA CELIA WHITE ROBAT EMULOV .59 Cumulative Subject Index Volumes 61-74, 76-80 Edited yb DRAWDE A. DENNIS DNA AHTRAM G. SINNED EMULOV .69 Biomembranes Part J: Membrane Biogenesis: Assembly and Targeting (General Methods; Eukaryotes) Edited yb YENDIS REHCSIELF DNA ACCEB REHCSIELF EMULOV .79 Biomembranes Part K: Membrane Biogenesis: Assembly and Targeting (Prokaryotes, Mitochondria, and Chloroplasts) Edited by SIDNEY REHCSIELF DNA ACCEB REHCSIEEF EMULOV 98. Biomembranes (Part L: Membrane Biogenesis: Processing and Recycling) Edited yb SIDNEY REHCSIELF DNA ACCEB REHCSIELF EMULOV 99. Hormone Action (Part F: Protein Kinases) Edited by EIKCAJ D. NIBROC DNA JOEL G. NAMDRAH EMULOV 100. Recombinant DNA (Part B) Edited yb YAR Wu, ECNERWAL ,NAMSSORG DNA KIVIE EVADEOM EMULOV 101. Recombinant DNA (Part )C Edited yb YAR Wu, ECNERWAL ,NAMSSORG DNA KIVIE EVADLOM EMULOV 102. Hormone Action (Part G: Calmodulin and Calcium-Binding Pro- teins) Edited yb YNOHTNA R. SNAEM DNA BERT W. O'MALLEY EMULOV 103. Hormone Action (Part H: Neuroendocrine Peptides) Edited yb P. LEAHCIM NNOC EMULOV 104. Enzyme Purification and Related Techniques (Part C) Edited yb WILLIAM B. YBOKAJ EMULOV 105. Oxygen Radicals in Biological Systems Edited by RETSEL REKCAP EMULOV 106. Posttranslational Modifications (Part A) Edited yb FINN WOLD DNA KIVIE EVADLOM EMULOV 107. Posttranslational Modifications (Part B) Edited by FINN WOLD DNA KIVIE EVADEOM EMULOV 108. Immunochemical Techniques (Part G: Separation and Character- ization of Lymphoid Cells) Edited yb INNAVOIG DI ,OTABAS JOHN J. ,ENOGNAL DNA NELEH NAV SIKANUV