Sulfur and Sulfur Amino Acids

647 Pages·1987·31.25 MB·English

by  William B. Jakoby

Contributors to Volume 143
Article numbers are in parentheses lbllowing the names of contributors.
Affiliations listed are current,
DOUGLAS 0. ADAMS (72), Department of PETER K. CHIANG (34, 64), Division of -oiB
Viticulture and Enology, University of chemistry, Walter Reed Army Institute of
,ainr?ifilaC Davis, Cahfornia 61659 Research, Washington, D.C. 70302
HOWARD ADLER (10), Department of Bio- JOHN D. H. COOPER (22), Department of
chemistry and Molecular Biology, ehT Biochemistry, Coventry and Warwick-
University of Chicago, Chicago, Illinois shire Hospital, Coventry CVI 4FH, -nE
73606 gland
TERESA ALONSO (34), Department of Bio- EUGENE G. DEMASTER (17), Medical Re-
chemistry, Walter Reed Army Institute of search Laboratories, Veterans Adminis-
Research, Washington, D.C. 20307 tration Medical Center, Minneapolis,
MARY E. ANDERSON (57), Department of Minnesota 55417
Biochemistry, Cornell University Medical JAMES DE LA ROSA (26), Division of Nutri-
College, New York, New York 12001 tional Sciences, Cornell University,
WILLIAM M. AWAD, JR. (65), Departments Ithaca, New York 35841
of Medicine and Biochemistry, University MICHAEL W. DUFFEL (25), Department tf
of Miami School of Medicine, Miami, Medicinal Chemistry and Natural Prod-
Florida 10133 ucts, College of Pharmacy, University of
DAVID H. BAKER (55), Department of Ani- Iowa, Iowa City, Iowa 24225
real Sciences and Division of Nutritional NOBUYOSHI ESAKI (24, 45, 54, 70, 79, 86),
Sciences, University of Illinois, Urbana, Institute for Chemical Research, Kyoto
Illinois 10816 University, Uji, Kyoto-Fu 611, Japan
HANS-OTTO BEUTLER (3), Boehringer- ROBERT C. FAHEY (14, 15), Department of
Mannheim GmbH, Biochemical Research Chemistry, University of California-San
Center, 2318-D Tutzing, Federal Republic Diego, La Jolla, ainrbJilaC 39029
of Germany HEINZ FANKHAUSER (61), CIBA-Geigy
RAYMOND F. BURK (56), Department of Ltd., CH-4002 Basel, Switzerland
Medicine, University of Texas Health Sci- MARC W. FARISS (16), Department of Pa-
ence Center, San Antonio, Texas 48287 thology, Medical College of Virginia,
Richmond, Virginia 89232
MARY R. BURROUS (42), Department of Bio-
chemistry and Molecular Biology, ehT JACK H. FELLMAN (30, 32, 39), Department
University of Chicago, Chicago, Illinois of Biochemistry, ehT Oregon Health Sci-
73606 ences University, Portland, Oregon 10279
ERNEST B. CAMPBELL (29, 51), Department S. J. FOSTER (35), Department of Nutri-
of Biochemistry, Cornell University Medi- tional Sciences, University "~ Wisconsin-
cal College, New York, New York 12001 Madison, Madison, Wisconsin 60735
THOMAS R. CHAUNCEY (60), Department of CLEMENT E. FURLONG (23), Division of
Biochemistry and Molecular Biology, ehT Medical Genetics, Center for Inherited
University of Chicago, Chicago, Illinois Diseases, University of Washington, Se-
73606 attle, Washington 55189
xiii
xiv CONTRIBUTORS TO VOLUME 143
HOWARD E. GANTHER (9, 35), Department KAZUHIRO 1MAI (12), Branch Hospital
of Nutritional Sciences, University of Pharmacy, University of Tokyo, Bunkyo-
Wisconsin-Madison, Madison, Wisconsin ku, Tokyo 112, Japan
60735 MAKOTO ISHIMOTO (40, 87), Department of
LEONARD J. GARBER (61), lnstitutefi)r Pho- Chemical Microbiology, Faculty of Phar-
tobiology, Brandeis University, Waltham, maceutical Sciences, Hokkaido Univer-
Massachusetts 45220 sity, Sapporo, Hokkaido 060, Japan
JOHN GIOVANELLI (71, 76), Laboratory of KIMIKAZU IWAMI (74), Department of Agri-
General and Comparative Biochemistry, cultural Chemistry, Kyoto Prefectural
Section on Alkaloid Biosynthesis, Na- University, Shimogamo Nakaragicho,
tional Institute of Mental Health, Be- Sakyoku, Kyoto 606, Japan
thesda, Maryland 21202 WILLIAM B. JAKOBY (36), Laboratory of
LOWELL A. GOLDSMITH (90), Dermatology Biochemistry and Metabolism, National
Unit, University of Rochester Medical Institute of Diabetes and Digestive and
Center, Rochester, New York 24641 Kidney Diseases, National Institutes of
RICHARD K. GORDON (34), Division of -DiB Health, Bethesda, Maryland 29802
chemistry, Walter Reed Army Institute of HIERONIM JAKUBOWSKI (73), Katedra Bio-
Research, Washington, D.C. 70302 chemii, Akademia Rolnicza, .IH Wolytiska
ALICE A. GREENE (23), Department of Pedi- ,53 PL-60-637 Poznah, Poland
atrics, University of California-San PETER C. JOCELYN (I ,1 46), Department of
Diego, La Jolla, CaliJbrnia 39029 Biochemistry, University of Edinburgh
Medical School, Edinburgh EH8 9XD,
OWEN W. GRIFFITH (29, 38, 49, 50, 51, 53,
63, 68), Department of Biochemistry, Cor- Scotland
nell University Medical College, New CARL R. JOHNSON (52), Department of
York, New York 12001 Chemistry, Wayne State University, De-
ANDRZEJ GURANOWSKI (73), Katedra Bio- troit, Michigan 20284
chemii, Akademia Rolnicza, .IH -oW HAROLD KADIN (47), Analytical Research
,akszFyl PL-60-637 PoznatL Poland and Development, The Squibb Institute
DAVID I. HALL (75), Department of Botany, for Medical Research, New Brunswick,
Ohio University, Athens, Ohio 10754 New Jers'ey 30980
PATRICK HARDEN (41), W. Alton Jones Cell HIROSH KANZAKI (85), Department of Ag-
Science Center, Lake Placid, New York ricultural Chemistry, Kyoto University,
64921 Kyoto 606, Japan
MICHAEL A. HAYWARD (51), Department of H. KONDO (40, 87), Department of Chemi-
Biochemistry, Cornell University Medical cal Microbiology, Faculty of Pharmaceu-
College, New York, New York 12001 tical Sciences, Hokkaido University, Sap-
LAWRENCE L. HIRSCHBERGER (26), Division poro, Hokkaido 060, Japan
of Nutritional Sciences, Cornell Univer- YASUO KONISHI (18), Monsanto Company
sity, Ithaca, New York 35841 AA4I, Chesterfield, Missouri 89136
H. ROBERT HORTON (89), Department of EDWARD M. KOSOWER (13, 48), School of
Biochemistry, North Carolina State Uni- Chemistry, Sackler Faculty f~c Exact Sci-
versity, Raleigh, North Carolina 59672 ences, TeI-Aviv University, Ramat-Aviv,
Yu HOSOKAWA (67), Division ~f Maternal leT Aviv ,87996 Israel
and Child Nutrition, National Institute of NECHAMA S. KOSOWER (13, 48), Depart-
Nutrition, Toyama, Shinjuku-ku, Tokyo ment of Human Genetics, Sackler School
,261 Japan of Medicine, Tel-Aviv University, Ramat-
JANETTE HOUK (21), Environmental Protec- Aviv, Tel Aciv ,87996 Israel
tion Agency, Washington, D.C. 06402 JAN P. KRAUS (66), Department ~fHuman
CONTRIBUTORS TO VOLUME 143 XV
Genetics, Yale University School of Medi- University of Texas at Austin, Austin,
cine, New Haven, Connecticut 01560 Texas 21787
R. J. KRAUS (9, 35), Department of Nutri- SENGODA G. RAMASWAMY (36), Laboratory
tional Sciences, University of Wisconsin- of Biochemistry and Metabolism, Na-
Madison, Madison, Wisconsin 60735 tional Institute of Diabetes and Digestive
K. KORIYAMA (28), Department of Pharma- and Kidney Diseases, National Institutes
cology, Kyoto Prefectural University of of Health, Bethesda, Maryland 29802
Medicine, Kawaramachi-Hirokaji, Kami- BETH REDFERN (17), Medical Research
myo-Ku, Kyoto 602, Japan Laboratories, Veterans Administration
JOHN LANE (42), Department of Biochemis- Medical Center, Minneapolis, Minnesota
try and Molecular Biology, The Univer- 71455
sity of Chicago, Chicago, Illinois 73606 DONALD J. REED (16), Department of -DiB
GERALD LARSEN (41), Metabolism and Ra- chemistry, Oregon State University, Cor-
diation Research Laboratory, U.S. De- vallis, Oregon 13379
partment of Agriculture, North Dakota FRANCO RENOSTO (59), Department of -DiB
State University Station, Fargo, North chemistry and Biophysics, University of
Dakota 50185 California, Davis, ainr~ifilaC 61659
FRANZ-JOSEF LHNWEBER (4, 27), Depart- RUSSELL B. RICHERSON (69), Abbott Labo-
ment of Drug Metabolism, Hoffman-La ratories, North Chicago, Illinois 46006
Roche, Inc., Nutley, New Jersey 01170 ALEXANDER B. YOR (37, 62), Protein Chem-
DANlEL J. LOGAN (25), Clayton Foundation istry Group, John Curtin School of Medi-
Biochemical Institute, The University ~f cal Research, Australian National Uni-
Texas at Austin, Austin, Texas 21787 versity, Canberra ACT 2601, Australia
MARJORIE F. Lou (20), Alcon Laboratories, TEKCHAND SAIDHA (58, 61), Institute for
Inc., Fort Worth, Texas 10167 Photobiology, Brandeis University, Wal-
JOHN H. MANGUM (65), Department of tham, Massachusetts 45220
Chemistry, Brigham Young University, VALENTINE H. SCHAEFFER (41), Depart-
Provo, Utah 20648 ment of Biochemistry and Biophysics,
ALTON MEISTER (57), Department of -DiB Center for Drugs and Biologics, Food and
chemistry, Cornell University Medical Drug Administration, Bethesda, Mary-
College, New York, New York 12001 land 29802
GEORGE A. MIuRA (34), Division of -DiB HAROLD A. SCHERAGA (18), Baker Labora-
chemistry, Walter Reed Army Institute of tory of Chemistry, Cornell University,
Research, Washington, D.C. 70302 Ithaca, New York 35841
KENNETH J. MONTY (4, 27), Department of JEROME A. SCHIFF (58, 61), Institute for
Biochemistry, University of Tennessee, Photobiology, Brandeis University, Wal-
Knoxville, Tennessee 69973 tham, Massachusetts 45220
TORU NAGASAWA (82, 85), Department of AHLERT SCHMIDT (77), Botanisches lnsti-
Agricultural Chemistry, Kyoto Univer- tat, Universitdt Miinchen, D-8000 Munich
sity, Kyoto 606, Japan ,91 Federal Republic of Germany
GERALD L. NEWTON (14, 15), Department JERRY A. SCHNEIDER (23), Department of
of Chemistry, University o( California- Pediatrics, University of California-San
San Diego, La Jolla, Califi)rnia 39029 Diego, La Jolla, California 39029
HACHIRO OZAKI (81), Central Research IRWIN H. SEGEL (59), Department of -DiB
Laboratories, Ajinomoto Co., Int., Ka- chemistry and Biophysics, University of
wasaki 210, Japan ,ainr)ifilaC Davis, California 61659
LAWRENCE L. POULSEN (20), Clayton PETER A. SEUBERT (59), Center for the Neu-
Foundation Biochemical Institute, The robiology of Learning and Memory, Uni-
xvi CONTRIBUTORS TO VOLUME 143
versity of California, lroine, California Laboratory of Chemistry, Cornell Univer-
71729 sity, Ithaca, New York 35841
ISAMU SHIIO (81), Central Research Labo- SEIZEN TOYAMA (88), Department of Agri-
ratories, Ajinomoto Co., Inc., Yokohama cultural Chemistry, University of the Ryu-
,442 Japan kyus, Nishihara, Okinawa ,10-309 Japan
R~EEVA SINGH (21), Department of Chem- TOSHIMASA TOYO'OKA (12), Department of
istry, Harvard University, Cambridge, Foods, National Institute of Hygienic Sci-
Massachusetts 83120 ences, Setagaya-ku, Tokyo ,851 Japan
WILLIAM E. SKIB~, (65), Department of -oiB D. C. TURNEEL (22), Department of Bio-
chemistry, University of Miami School of chemistry, Coventry and Warwickshire
Medicine, Miami, Florida 10133 Hospital, Coventry CVI 4FH, England
MARK X. SLIWKOWSKI (19), Triton Bio- TOSHIHIKO UBUKA (5), Department of Bio-
sciences, Inc., Alameda, California 10549 chemistry, Okayama University Medical
IVAN K. SMITH (75), Department of Botany, School, Okayama 700, Japan
Ohio University, Athens, Ohio 10754 LAWRENCE C. UHTEG (60), Department f.o
MARGARET SMITH (23), Department of -eP Biochemistry and Molecular Biology, ehT
diatries, University of California-San University of Chicago, Chicago, Illinois
Diego, La Jolla, California 39029 73606
KENJI SODA (24, 43, 44, 45, 54, 70, 78, 79, JACK R. UREN (84), Battellee Columbus
86, 88), Institute for Chemical Research, Laboratories, Columbus, Ohio 10234
Kyoto University, Uji, Kyoto-Fu 611, RANDOLPH T. WEDDING (8), Department of
Japan Biochemistry, University of California,
Bo $6R8O (1, 31), Department f.o Clinical Riverside, California 12529
Chemistry, LinkOping University, Re- CATHERINE L. WEINSTEIN (38, 49, 68), -eD
gional Hospital, LinkOping, Sweden partment of Biochemistry, Cornell Uni-
versity Medical College, New York, New
JAMES L. STEVENS (41), .W Alton Jones Cell
York 12001
Science Center, Lake Placid, New York
64921 MARILYN S. WELLS (65), Department of
Medicine, Veterans Administration Medi-
MARTHA H. STIPANUK (26), Division qfNu-
cal Center, Miami, Florida 33101, and
tritional Scienves, Cornel/ University,
University of Miami School of Medicine,
Ithaca, New York 35841
Miami, Florida 10133
DARRVL M. SULLIVAN (2), Hazleton Labo-
AIKO WESTLEY (42), Department of Bio-
ratories America, Inc., Madison, Wiscon-
chemistry and Molecular Biology, ehT
sin 50735
University of Chicago, Chicago, Illinois
HAROLD E. SWAISGOOD (19, 89), Depart- 73606
ment f~( Food Science and Biochemistry, JOHN WESTLEY (6, 10, 42, 60), Department
North Carolina State University, Raleigh, of Biochemistry and Molecular Biology,
North Carolina 59672 ehT University of Chicago, Chicago, Illi-
HIOEHIKO TANAKA (43, 44), Institute for nois 73606
Chemical Research, Kyoto University, MARK P. WESTRICK (52), Department of
Uji, Kyoto-Fu 611, Japan Chemistry, Wayne State University, -eD
Y. TANAKA (28), Department of Pharmacol- troit, Michigan 20284
ogy, Kyoto Prefectural University of GEORGE M. WmTESIDES (21), Department
Medicine, Kawaramachi-Hirokoji, Kyoto f.o Chemistry, Harvard University, Cam-
,206 Japan bridge, Massachusetts 83120
THEODORE W. THANNHAUSER (18), Baker ELISABETH M. WOLF-HEuss (33), Analyti-
CONTRIBUTORS OT VOLUME 143 xvii
cal Department, Degassa Pharma Group, IHSORIH AWAGANAY (91), Mitsubishi-Kasei
0006-D Frankfurt l, Federal Republic of Institute "q ,~~L Sciences, Maehida,
Germany Tokyo ,491 Japan
JOHN L. WOOD (7), Department of -DiB GNAHS FA YANG (72), Department of Vege-
chemistry, University of Tennessee-Mem- table Crops, University of ,ainr)ifilaC
phis, Memphis, Tennessee 11183 Davis, California 61659
HIDEAKI YAMADA (82, 85), Department of NEDOYK OTOMUSAY (74), Research Insti-
Agricultural Chemistry, Kyoto Univer- tute fin" Food Science, Kyoto University,
sity, Kyoto 606, Japan Gokasho, Uji, Kyoto 611, Japan
SHuzo ATAGAMAY (80, 83), Department of KAZUO YONAHA (88), Department of Agri-
Biology, Facahy of General Education, cultural Chemistry, University of the Ryu-
h~G University, Gfu ,11-105 Japan kyus, Nishihara, Okinawa ,10-309 Japan
KENJI IHCUGAMAY (67), Division of Mater- DANIEL M. ZIEGLER (20, 25, 69j, Clayton
nal and Child Nutrition, National Insti- Foundation Biochemical Institute, ehT
tute of Nutrition, Toyama, Shinjuku-ku, University of Texas at Austin, Austin,
Tokyo ,261 Japan Texas 21787
Preface
Because of the large number of diverse and critical roles played by
sulfur in several of its oxidation states, the subject of testing and prepar-
ing derivatives of sulfur or of measuring, purifying, and characterizing
enzymes associated with sulfur metabolism is represented in most of the
individual volumes of Methods ni Enzymology. For example, many of the
methyl transferases which utilize S-adenosylmethionine have found a
home in volumes that deal with nucleic acids, lipids, amines, or detoxica-
tion as the major topic; the polyamines are treated in a separate volume.
Enzymes and procedures involving sulfur-containing metabolites such as
coenzyme A are presented in most of the volumes dealing with metabolic
enzymes, and aspects of biotin biochemistry have appeared in books in
this series which range in emphasis from vitamins to affinity labeling.
Since no single book could cover all of these topics in a reasonable man-
ner, such dispersal is entirely appropriate. In fact, most of the examples
cited above are not presented in this volume. It is because of the broad
distribution of topics that we include, as an appendix, a brief index to
other articles on some of the subject areas covered here.
Progress and interest in the work on sulfur and sulfur amino acids, on
seleno and tellurium analogs, and on the separation and quantitation of
sulfur-containing compounds have led to the suggestion that a single
source book of methods would be useful. We have responded by provid-
ing descriptions of a large number of analytical methods for inorganic and
organic sulfur compounds in their several oxidation states. This volume
also includes general and specific methods of preparation for a number of
organic sulfur compounds and presents procedures for the purification
and assay of a group of enzymes that participate in inorganic sulfur me-
tabolism as well as in the pathways of formation and utilization of cys-
teine, methionine, and related compounds.
We express our appreciation to the individual authors who have con-
tributed to the volume and, in the process, have made their often unique
expertise available to other investigators interested in the biochemistry of
sulfur.
WILLIAM B. JAKOBY
OWEN W. GRIFFITH
xix
METHODS IN ENZYMOLOGY
EDITED YB
Sidney P. Colowick and Nathan O. Kaplan
VANDERBILT UNIVERSITY DEPARTMENT OF CHEMISTRY
SCHOOL OF MEDICINE UNIVERSITY OF CALIFORNIA
NASHVILLE, TENNESSEE AT SAN DIEGO
LA JOLLA. CALIFORNIA
.1 Preparation and Assay of Enzymes
II. Preparation and Assay of Enzymes
III. Preparation and Assay of Substrates
IV. Special Techniques for the Enzymologist
.V Preparation and Assay of Enzymes
VI. Preparation and Assay of Enzymes )deunitnoC(
Preparation and Assay of Substrates
Special Techniques
VII. Cumulative Subject Index
xxi
METHODS IN ENZYMOLOGY
EDITORS-IN-CHIEF
Sidney P. Colowick and Nathan O. Kaplan
EMULOV VIII. Complex Carbohydrates
Edited by ELIZABETH F. NEUFELD DNA VICTOR GINSBURG
EMULOV IX. Carbohydrate Metabolism
Edited by WILLIS A. DOOW
EMULOV X. Oxidation and Phosphorylation
Edited yb DLANOR W. AND MAYNARD ESTABROOK E. PULLMAN
EMULOV XI. Enzyme Structure
Edited by C. H. W. HIRS
EMULOV XII. Nucleic Acids (Parts A and B)
Edited yb LAWRENCE AND GROSSMAN KIVIE MOLDAVE
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
VOLUME XVI. Fast Reactions
Edited yb KENNETH NITSUK
EMULOV XVII. Metabolism of Amino Acids and Amines (Parts A and B)
Edited by HERBERT TABOR AND CELIA WHITE TABOR
iiixx
xxiv SDOHTEM NI YGOLOMYZNE
EMULOV XVIII. Vitamins and Coenzymes (Parts A, B, and C)
Edited yb DLANOD B. KCIMROCCM DNA LEMUEL D. THG1RW
VOLUME XlX. Proteolytic Enzymes
Edited yb GERTRUDE E. PERLMANN AND LASZLO LORAND
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 B. WILLIAM YBOKAJ
EMULOV XXIII. Photosynthesis (Part A)
Edited yb YNOHTNA NAS ORTEIP
EMULOV XXIV. Photosynthesis and Nitrogen Fixation (Part B)
Edited yb YNOHTNA NAS ORTE1P
EMULOV XXV. Enzyme Structure (Part B)
Edited yb C. H. W. HIRS DNA EGRES N. FFEHSAM1T
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 GRUaSNIG
EMULOV XXIX. Nucleic Acids and Protein Synthesis (Part E)
Edited yb ECNERWAL NAMSSORG DNA KIVlE EVADLOM
EMULOV XXX. Nucleic Acids and Protein Synthesis (Part F)
Edited yb KIVIE EVADLOM DNA ECNERWAL NAMSSORG
EMULOV XXXI. Biomembranes (Part A)
Edited by REHCSIELF YENDIS DNA RETSEL REKCAP
METHODS IN ENZYMOLOGY XXV
EMULOV XXXII. Biomembranes (Part B)
Edited by YENDIS REHCSIELF DNA RETSEL REKCAP
EMULOV XXXIII. Cumulative Subject Index Volumes I-XXX
Edited by 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
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. NAMDRAH DNA BERT W. O'MALLEY
EMULOV XXXIX. Hormone Action (Part D: Isolated Cells, Tissues, and
Organ Systems)
Edited yb JOEL G. NAMDRAH DNA BERT W. O'MALLEV
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 by KLAUS HCABSOM
EMULOV XLV. Proteolytic Enzymes (Part B)
Edited yb LASZLO DNAROL