METHODS IN ENZYMOLOGY EDITORS-IN-CHIEF John N. Abelson Melvin I. Simon DIVISION OF BIOLOGY CALIFORNIA INSTITUTE OF TECHNOLOGY PASADENA, CALIFORNIA FOUNDING EDITORS Sidney P. Colowick and Nathan O. Kaplan Contributors to Volume 378 Article numbers are in parentheses and following the names of contributors. Affiliations listed are current. Kotb Abdelmohsen (20), Institut fu ¨ r Biochemie und Molekularbiologie I, Heinrich-Heine-Universita ¨ tDu ¨ sseldorf, D-40225 Du ¨ sseldorf, Germany Antonio Arroyo (17), Centro Andaluz de Biologı ´ a del Desarrollo, Universidad Pablo de Olavide, E-41013 Sevilla, Spain Maurizio Battino (12, 14), Institute of Biochemistry, Universita ` Politecnica delle Marche, 60131 Ancona, Italy Mikael Bjo ¨ rnstedt (8), Division of Pathology, Department of Laboratory Medicine, Karolinska Institutet, Hud- dinge University Hospital, SE-141 86 Stockholm, Sweden Judy L. Bolton (6), Department of Med- icinal Chemistry and Pharmacognosy, College of Pharmacy, University of Illinois at Chicago, Chicago, Illinois 60612-7231 Stefano Bompadre (12, 14), Institute of Microbiology and Biomedical Sciences, Universita ` Politecnica delle Marche, 60131 Ancona, Italy Gaetano Cairo (25), Institute of General Pathology, University of Milan School of Medicine, 20133 Milan, Italy Christopher R. Chitambar (26), Division of Neoplastic Diseases, Medical College of Wisconsin, Milwaukee, Wisconsin 53226 Tadeusz Chojnacki (11), Institute of Bio- chemistry and Biophysics, Polish Acad- emy of Sciences, 02-106 Warszawa, Poland Gianfranca Corna (25), Institute of Gen- eral Pathology, University of Milan School of Medicine, 20133 Milan, Italy Gustav Dallner (1), Department of Bio- chemistry and Biophysics, Stockholm University, S-106 91, Stockholm, Sweden Dale A. Dickinson (23, 24), Department of Environmental Health Sciences, School of Public Health, and Center for Free Radical Biology, University of Alabama at Birmingham, Birmingham, Alabama 35294 Marta Dubin (4), Bioenergetics Research Centre, National Research Council, School of Medicine, University of Buenos Aires, 1121-Buenos Aires, Argentine Jennifer L. DuBois (2), Department of Chemistry, University of California, Berkeley, Berkeley, California 94720 Daniele Fattorini (14), Institute of Biol- ogy and Genetics, Universita ` Politecni- ca delle Marche, 60131 Ancona, Italy Silvia Ferna ´ ndez Villamil (4), Bioener- getics Research Centre, National Re- search Council, School of Medicine, University of Buenos Aires, 1121- Buenos Aires, Argentine Henry Jay Forman (23, 24), Department of Environmental Health Sciences, School of Public Health, and Center for Free Radical Biology, University of Alabama at Birmingham, Birmingham, Alabama 35294* *Current affiliation: Division of Natural Sciences, University of California, Merced, Merced, California 95344 xi Craig A. Gay (13), Centre for Vascular Research, School of Medical Sciences, University of New South Wales, Syd- ney, New South Wales 2052, Australia Jacob Gru ¨ nler (1), Department of Mole- cular Medicine, Karolinska Hospital, Karolinska Institutet Medical School, S-106 91, Stockholm, Sweden H C. Huang (22), Schering-Plough Research Institute, Kenilworth, New Jersey 07033-1300 Karen E. Iles (23), Department of Envir- onmental Health Sciences, School of Public Health, and Center for Free Radical Biology, University of Alabama at Birmingham, Birmingham, Alabama 35294 Anil K. Jaiswal (18), Department of Pharmacology, Baylor College of Medi- cine, Houston, Texas 77030 Jeffrey A. Johnson (19), University of Wisconsin-Madison, School of Phar- macy, Madison, Wisconsin 53705-2222 Delinda A. Johnson (19), University of Wisconsin-Madison, School of Phar- macy, Madison, Wisconsin 53705-2222 Joy Joseph (26), Biophysics Department, Medical College of Wisconsin, Milwau- kee, Wisconsin 53226 Shasi V. Kalivendi (26), Biophysics De- partment, Medical College of Wiscon- sin, Milwaukee, Wisconsin 53226 B. Kalyanaraman (26), Biophysics De- partment, Medical College of Wiscon- sin, Milwaukee, Wisconsin 53226 Sergey Kamzalov (9), Department of Molecular Pharmacology and Toxi- colgy, University of Southern Califor- nia, Los Angeles, California 90089-9121 Judith P. Klinman (2), Department of Chemistry, University of California, Berkeley, Berkeley, California 94720 Lars-Oliver Klotz (20), Institut fu ¨ r Biochemie und Molekularbiologie I, Heinrich-Heine-Universita ¨ tDu ¨ sseldorf, D-40225 Du ¨ sseldorf, Germany Akira Kobayashi (21), Center for Tsuku- ba Advanced Research Alliance, Insti- tute of Basic Medical Sciences and JST-ERATO Environmental Response Project, University of Tsukuba, Tsukuba 305-8577, Japan Eugene Konorev (26), Biophysics De- partment, Medical College of Wiscon- sin, Milwaukee, Wisconsin 53226 Srigiridhar Kotamraju (26), Biophysics Department, Medical College of Wis- consin, Milwaukee, Wisconsin 53226 Edward J. Land (5), School of Chemistry and Physics, Keele University, Keele, Staffordshire ST5 5BG, United Kingdom Jong-Min Lee (19), University of Wiscon- sin-Madison, School of Pharmacy, Madison, Wisconsin 53705-2222 Luciana Leone (12), Institute of Micro- biology and Biomedical Sciences, Uni- versita ` Politecnica delle Marche, 60100 Ancona, Italy Jiang Li (19), University of Wisconsin- Madison, School of Pharmacy, Madison, Wisconsin 53705-2222 Gian Paolo Littarru (14), Institute of Biochemistry, Universita ` Politecnica delle Marche, 60131 Ancona, Italy Rui-Ming Liu (24), Department of Envir- onmental Health Sciences, School of Public Health, and Center for Free Ra- dical Biology, University of Alabama, Birmingham, Alabama 35294 Ira Melchheier (20), Institut fu ¨ r Bio- chemie und Molekularbiologie I, Hein- rich-Heine-Universita ¨ tDu ¨ sseldorf, D-40225 Du ¨ sseldorf, Germany xii contributors to volume 378 Pierantonio Menna (25), Department of Drug Sciences and Centro Studi Invecchiamento, G. d’Annunzio Uni- versity School of Medicine, 66013 Chieti, Italy Giorgio Minotti (25), Department of Drug Sciences and Centro Studi Invec- chiamento, G. d’Annunzio University School of Medicine, 66013 Chieti, Italy D. James Morre ´ (15), Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, Lafayette, Indiana 47907 Fabrizio Mosca (14), Institute of Bio- chemistry, Universita ` Politecnica delle Marche, 60131 Ancona, Italy Pla ´ cido Navas (16, 17), Centro Andaluz de Biologı ´ a del Desarrollo, Universidad Pablo de Olavide, E-41013 Sevilla, Spain Truyen Nguyen (22), Schering-Plough Research Institute, Kenilworth, New Jersey 07033-1300 Tomas Nordman (8), Division of Path- ology, Department of Laboratory Medicine, Karolinska Institutet, Hud- dinge University Hospital, SE-141 86 Stockholm, Sweden Tsutomu Ohta (21), Genomics Division, National Cancer Center Resarch Insti- tute, Tokyo 104-0045, Japan Jerker M. Olsson (8), Division of Pathol- ogy, Department of Laboratory Medi- cine, Karolinska Institutet, Huddinge University Hospital, SE-141 86 Stock- holm, Sweden Pauline Patak (20), Institut fu ¨ r Biochem- ie und Molekularbiologie I, Heinrich- Heine-Universita ¨ tDu ¨ sseldorf, D-40225 Du ¨ sseldorf, Germany Trevor M. Penning (3), Department of Pharmacology, University of Pennsylva- nia School of Medicine, Philadelphia, Pennsylvania 19104-6084 Cecil B. Pickett (22), Schering-Plough Research Institute, Kenilworth, New Jersey 07033-1300 Christopher A. Ramsden (5), School of Chemistry and Physics, Keele Univer- sity, Keele, Staffordshire ST5 5BG, United Kingdom Igor Rebrin (9), Department of Molecu- lar Pharmacology and Toxicology, Uni- versity of Southern California, Los Angeles, California 90089-9121 Stefania Recalcati (25), Institute of General Pathology, University of Milan School of Medicine, 20133 Milan, Italy Patrick A. Riley (5), Gray Cancer Insti- tute, Mount Vernon Hospital, North- wood HA6 2JR, United Kingdom Juan C. Rodrı ´ guez-Aguilera (17), Cen- tro Andaluz de Biologı ´ a del Desarrollo, Universidad Pablo de Olavide, E-41013 Sevilla, Spain Emanuela Salvatorelli (25), Depart- ment of Drug Sciences and Centro Studi Invecchiamento, G. d’Annunzio Uni- versity School of Medicine, 66013 Chieti, Italy Carlos Santos-Ocan ˜ a (17), Centro An- daluz de Biologı ´ a del Desarrollo, Uni- versidad Pablo de Olavide, E-41013 Sevilla, Spain Philip J. Sherratt (22), Schering-Plough Research Institute, Kenilworth, New Jersey 07033-1300 Helmut Sies (20), Institut fu ¨ r Biochemie und Molekularbiologie I, Heinrich- Heine-Universita ¨ tDu ¨ sseldorf, D-40225 Du ¨ sseldorf, Germany Rajindar S. Sohal (9, 10), Department of Pharmacology and Toxicology, Univer- sity of Southern California, Los An- geles, California 90089-9121 contributors to volume 378 xiii Roland Stocker (13), Centre for Vas- cular Research, School of Medical Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia Andre ´ s O. M. Stoppani (4), Bioenergetics Research Centre, National Research Council, School of Medicine, University of Buenos Aires, 1121-Buenos Aires, Argentine Ewa Swiezewska (7), Institute of Bio- chemistry and Biophysics, Polish Acad- emy of Sciences, 02-106 Warszawa, Poland Gregory R. J. Thatcher (6), Department of Medicinal Chemistry and Pharma- cognosy, College of Pharmacy, Univer- sity of Illinois at Chicago, Chicago, Illinois 60612-7231 Jose ´ Manuel Villalba (16, 17), Departa- mento de Biologı ´ a Celular, Fisiologı ´ ae Inmunologı ´ a, Universidad de Co ´ rdoba, E-14071 Co ´ rdoba, Spain Claudia von Montfort (20), Institut fu ¨ r Biochemie und Molekularbiologie I, Heinrich-Heine-Universita ¨ tDu ¨ sseldorf, D-40225 Du ¨ sseldorf, Germany Nobuo Watanabe (24), Department of Environmental Health Sciences, School of Public Health, and Center for Free Radical Biology, University of Alabama, Birmingham, Alabama 35294 Amanda F. Wigley (23), Department of Environmental Health Sciences, School of Public Health, University of Alaba- ma at Birmingham, Birmingham, Ala- bama 35294* Masayuki Yamamoto (21), Center for Tsukuba Advanced Research Alliance, Institute of Basic Medical Sciences and JST-ERATO Environmental Response Project, University of Tsukuba, Tsuku- ba 305-8577, Japan Linning Yu (6), Department of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, University of Illinois at Chicago, Chicago, Illinois 60612-7231 *Current affiliation: Department of Pathology, University of Alabama at Birmingham, Birmingham, Alabama 35294 xiv contributors to volume 378 Preface Developments in genomics and proteomics rapidly generated focus on new -omics, particularly metabolomics and phenomics. Quinones, hydroquinones, semiquinones, and their metabolites are naturally-occurring compounds that serve as wonderful examples for this new paradigm of interdigitating -omics; in addition to a role as substrates and products in metabolism, quinone com- pounds are intermediates in many pathways of gene regulation, enzyme protein induction, feedback control, and waste product elimination. Quinones play a pivotal role in energy metabolism (Peter Mitchell’s protonmotive ‘‘Q cycle’’), many other key processes, and even in chemotherapy, where redox cycling drugs are utilized. The present volume of Methods in Enzymology on quinones and quinone enzymes serves to bring together current methods and concepts on this topic. It focuses on the role in the so-called Phase II of drug metabolism (xenobiotics), but includes aspects on Phase I (CYP, cytochromes P-450) and Phase III (transport systems) as well. This volume of Methods in Enzymology (Part A) focuses on quinones and quinone enzymes in terms of coenzyme Q (detection and quinone reductases), plasma membrane quinone reductases, and the role of quinones in cellular signaling and modulation of gene expression, whereas Part B addresses mitochondrial ubiquinone and reductases, anticancer qui- nones, and the role of quinone reductases in chemoprevention and nutrition, as well as the role of quinones in age-related diseases. Phase II Enzymes, Part C, will focus on glutathione, glutathione S-transferases, and other conjugation enzymes. The enzyme, NAD(P)H: quinone oxidoreductase, is the subject of a major section in this volume. This enzyme, discovered in 1958 in Stockholm by Lars Ernster and named DT-Diaphorase by him, has multiple roles, some of which were only recently discovered. There are human polymorphisms in these enzymes relating to variations in cancer risk, and enzymes targeted by quinones are being investigated. Modern methods in assaying quinone reactions and, indeed, various quinones themselves, are also included in this volume. Ubiquinone (Coenzyme Q 10 ) as a major naturally-occurring quinone became a highlight of scientific interest following its discovery in 1957, and was dis- covered to have an established role in mitochondrial electron transport by Frederick Crane. Fundamental contributions were made by Karl Folkers on xv its supplemental use for health benefits in disease prevention and by Andre ´ sO. M. Stoppani, a pioneer of Argentinian biochemistry, in utilizing quinones for the treatment of Chagas disease. We thank the Advisory Committee (Enrique Cadenas, Los Angeles; Gustav Dallner, Stockholm; Tom Kensler, Baltimore; Lars-Oliver Klotz, Du ¨ sseldorf; David Ross, Denver) for their valuable suggestions and wisdom in selecting the contributions for this volume. Helmut Sies Lester Packer xvi preface METHODS IN ENZYMOLOGY Volume I. Preparation and Assay of Enzymes Edited by Sidney P. Colowick and Nathan O. Kaplan Volume II. Preparation and Assay of Enzymes Edited by Sidney P. Colowick and Nathan O. Kaplan Volume III. Preparation and Assay of Substrates Edited by Sidney P. Colowick and Nathan O. Kaplan Volume IV. Special Techniques for the Enzymologist Edited by Sidney P. Colowick and Nathan O. Kaplan Volume V. Preparation and Assay of Enzymes Edited by Sidney P. Colowick and Nathan O. Kaplan Volume VI. Preparation and Assay of Enzymes (Continued) Preparation and Assay of Substrates Special Techniques Edited by Sidney P. Colowick and Nathan O. Kaplan Volume VII. Cumulative Subject Index Edited by Sidney P. Colowick and Nathan O. Kaplan Volume VIII. Complex Carbohydrates Edited by Elizabeth F. Neufeld and Victor Ginsburg Volume IX. Carbohydrate Metabolism Edited by Willis A. Wood Volume X. Oxidation and Phosphorylation Edited by Ronald W. Estabrook and Maynard E. Pullman Volume XI. Enzyme Structure Edited by C. H. W. Hirs Volume XII. Nucleic Acids (Parts A and B) Edited by Lawrence Grossman and Kivie Moldave Volume XIII. Citric Acid Cycle Edited by J. M. Lowenstein Volume XIV. Lipids Edited by J. M. Lowenstein Volume XV. Steroids and Terpenoids Edited by Raymond B. Clayton xvii Volume XVI. Fast Reactions Edited by Kenneth Kustin Volume XVII. Metabolism of Amino Acids and Amines (Parts A and B) Edited by Herbert Tabor and Celia White Tabor Volume XVIII. Vitamins and Coenzymes (Parts A, B, and C) Edited by Donald B. McCormick and Lemuel D. Wright Volume XIX. Proteolytic Enzymes Edited by Gertrude E. Perlmann and Laszlo Lorand Volume XX. Nucleic Acids and Protein Synthesis (Part C) Edited by Kivie Moldave and Lawrence Grossman Volume XXI. Nucleic Acids (Part D) Edited by Lawrence Grossman and Kivie Moldave Volume XXII. Enzyme Purification and Related Techniques Edited by William B. Jakoby Volume XXIII. Photosynthesis (Part A) Edited by Anthony San Pietro Volume XXIV. Photosynthesis and Nitrogen Fixation (Part B) Edited by Anthony San Pietro Volume XXV. Enzyme Structure (Part B) Edited by C. H. W. Hirs and Serge N. Timasheff Volume XXVI. Enzyme Structure (Part C) Edited by C. H. W. Hirs and Serge N. Timasheff Volume XXVII. Enzyme Structure (Part D) Edited by C. H. W. Hirs and Serge N. Timasheff Volume XXVIII. Complex Carbohydrates (Part B) Edited by Victor Ginsburg Volume XXIX. Nucleic Acids and Protein Synthesis (Part E) Edited by Lawrence Grossman and Kivie Moldave Volume XXX. Nucleic Acids and Protein Synthesis (Part F) Edited by Kivie Moldave and Lawrence Grossman Volume XXXI. Biomembranes (Part A) Edited by Sidney Fleischer and Lester Packer Volume XXXII. Biomembranes (Part B) Edited by Sidney Fleischer and Lester Packer Volume XXXIII. Cumulative Subject Index Volumes I-XXX Edited by Martha G. Dennis and Edward A. Dennis Volume XXXIV. Affinity Techniques (Enzyme Purification: Part B) Edited by William B. Jakoby and Meir Wilchek xviii methods in enzymology Volume XXXV. Lipids (Part B) Edited by John M. Lowenstein Volume XXXVI. Hormone Action (Part A: Steroid Hormones) Edited by Bert W. O’Malley and Joel G. Hardman Volume XXXVII. Hormone Action (Part B: Peptide Hormones) Edited by Bert W. O’Malley and Joel G. Hardman Volume XXXVIII. Hormone Action (Part C: Cyclic Nucleotides) Edited by Joel G. Hardman and Bert W. O’Malley Volume XXXIX. Hormone Action (Part D: Isolated Cells, Tissues, and Organ Systems) Edited by Joel G. Hardman and Bert W. O’Malley Volume XL. Hormone Action (Part E: Nuclear Structure and Function) Edited by Bert W. O’Malley and 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 Mosbach Volume XLV. Proteolytic Enzymes (Part B) Edited by Laszlo Lorand Volume XLVI. Affinity Labeling Edited by William B. Jakoby and Meir Wilchek Volume XLVII. Enzyme Structure (Part E) Edited by C. H. W. Hirs and Serge N. Timasheff Volume XLVIII. Enzyme Structure (Part F) Edited by C. H. W. Hirs and Serge N. Timasheff Volume XLIX. Enzyme Structure (Part G) Edited by C. H. W. Hirs and Serge N. Timasheff Volume L. Complex Carbohydrates (Part C) Edited by Victor Ginsburg Volume LI. Purine and Pyrimidine Nucleotide Metabolism Edited by Patricia A. Hoffee and Mary Ellen Jones Volume LII. Biomembranes (Part C: Biological Oxidations) Edited by Sidney Fleischer and Lester Packer Volume LIII. Biomembranes (Part D: Biological Oxidations) Edited by Sidney Fleischer and Lester Packer methods in enzymology xix [...]... P450 (Part B) Edited by Eric F Johnson and Michael R Waterman Volume 273 RNA Polymerase and Associated Factors (Part A) Edited by Sankar Adhya Volume 274 RNA Polymerase and Associated Factors (Part B) Edited by Sankar Adhya Volume 275 Viral Polymerases and Related Proteins Edited by Lawrence C Kuo, David B Olsen, and Steven S Carroll Volume 276 Macromolecular Crystallography (Part A) Edited by Charles... RNA Polymerases and Associated Factors (Part C) Edited by Sankar L Adhya and Susan Garges Volume 371 RNA Polymerases and Associated Factors (Part D) Edited by Sankar L Adhya and Susan Garges Volume 372 Liposomes (Part B) ¨ ¨ Edited by Negat Duzgunes, Volume 373 Liposomes (Part C) ¨ ¨ Edited by Negat Duzgunes, Volume 374 Macromolecular Crystallography (Part D) Edited by Charles W Carter, Jr., and Robert... 269 Nitric Oxide (Part B: Physiological and Pathological Processes) Edited by Lester Packer Volume 270 High Resolution Separation and Analysis of Biological Macromolecules (Part A: Fundamentals) Edited by Barry L Karger and William S Hancock Volume 271 High Resolution Separation and Analysis of Biological Macromolecules (Part B: Applications) Edited by Barry L Karger and William S Hancock Volume 272... Sweet Volume 375 Chromatin and Chromatin Remodeling Enzymes (Part A) Edited by C David Allis and Carl Wu Volume 376 Chromatin and Chromatin Remodeling Enzymes (Part B) Edited by C David Allis and Carl Wu Volume 377 Chromatin and Chromatin Remodeling Enzymes (Part C) Edited by C David Allis and Carl Wu Volume 378 Quinones and Quinone Enzymes (Part A) Edited by Helmut Sies and Lester Packer Volume 379 Energetics... Regulators (Part B: Calcium and Lipids) Edited by P Michael Conn and Anthony R Means Volume 142 Metabolism of Aromatic Amino Acids and Amines Edited by Seymour Kaufman Volume 143 Sulfur and Sulfur Amino Acids Edited by William B Jakoby and Owen Griffith Volume 144 Structural and Contractile Proteins (Part D: Extracellular Matrix) Edited by Leon W Cunningham Volume 145 Structural and Contractile Proteins (Part. .. Vesicular Traffic) Edited by W E Balch, Channing J Der, and Alan Hall Volume 330 Hyperthermophilic Enzymes (Part A) Edited by Michael W W Adams and Robert M Kelly Volume 331 Hyperthermophilic Enzymes (Part B) Edited by Michael W W Adams and Robert M Kelly Volume 332 Regulators and Effectors of Small GTPases (Part F: Ras Family I) Edited by W E Balch, Channing J Der, and Alan Hall Volume 333 Regulators and. .. by Nejat Duzgunes, Volume 222 Proteolytic Enzymes in Coagulation, Fibrinolysis, and Complement Activation (Part A: Mammalian Blood Coagulation Factors and Inhibitors) Edited by Laszlo Lorand and Kenneth G Mann Volume 223 Proteolytic Enzymes in Coagulation, Fibrinolysis, and Complement Activation (Part B: Complement Activation, Fibrinolysis, and Nonmammalian Blood Coagulation Factors) Edited by Laszlo... Carbohydrates (Part D) Edited by Victor Ginsburg Volume 84 Immunochemical Techniques (Part D: Selected Immunoassays) Edited by John J Langone and Helen Van Vunakis Volume 85 Structural and Contractile Proteins (Part B: The Contractile Apparatus and the Cytoskeleton) Edited by Dixie W Frederiksen and Leon W Cunningham Volume 86 Prostaglandins and Arachidonate Metabolites Edited by William E M Lands and. .. Interactions Edited by Jonathan B Chaires and Michael J Waring Volume 341 Ribonucleases (Part A) Edited by Allen W Nicholson Volume 342 Ribonucleases (Part B) Edited by Allen W Nicholson Volume 343 G Protein Pathways (Part A: Receptors) Edited by Ravi Iyengar and John D Hildebrandt Volume 344 G Protein Pathways (Part B: G Proteins and Their Regulators) Edited by Ravi Iyengar and John D Hildebrandt... Cytochrome P450, Part C Edited by Eric F Johnson and Michael R Waterman Volume 358 Bacterial Pathogenesis (Part C: Identification, Regulation, and Function of Virulence Factors) Edited by Virginia L Clark and Patrik M Bavoil Volume 359 Nitric Oxide (Part D) Edited by Enrique Cadenas and Lester Packer Volume 360 Biophotonics (Part A) Edited by Gerard Marriott and Ian Parker Volume 361 Biophotonics (Part B) Edited . Colowick and Nathan O. Kaplan Volume V. Preparation and Assay of Enzymes Edited by Sidney P. Colowick and Nathan O. Kaplan Volume VI. Preparation and Assay of Enzymes (Continued) Preparation and Assay. Free Radical Biology, University of Alabama, Birmingham, Alabama 35294 Amanda F. Wigley (23), Department of Environmental Health Sciences, School of Public Health, University of Alaba- ma at Birmingham,. School of Public Health, and Center for Free Radical Biology, University of Alabama at Birmingham, Birmingham, Alabama 35294 Anil K. Jaiswal (18), Department of Pharmacology, Baylor College of