Atlas of Protein Spectra in the Ultraviolet and Visible Regions Volume Edited by Donald M Kirschenbaum, Ph D Associate Professor Department of Biochemistry College of Medicine and School of Graduate Studies Downstate Medical Center State University of New York Brooklyn, New York IFI/PLENUM • NEWYORK-WASHINGTON-LONDON Library of Congress Catalog Card Number 77-183566 ISBN 0-306-67302-9 © 1974 IFI/Plenum Data Corporation A Subsidiary of Plenum Publishing Corporation 227 West 17th Street, New York, N.Y 10011 United Kingdom edition published by Plenum Press, London A Division of Plenum Publishing Company, Ltd Davis House (4th Floor), Scrubs Lane, Harlesden, NWlO 6SE, London, England All rights reserved No part of this publication may be reproduced in any form without written permission from the publisher Printed in the United States of America To Roslyn ACKNOWLEDGMENTS 11 He who ascribes a citation unto its author brings redemption into the world." Ethics of the Fathers, Megilla 15, Rabbi Elazar citing Rabbi Hanina Permission to publish the figures used in this book has been obtained from the editors, publishers, and, where necessary, authors, of the material cited in the following journals and books: Journals Acta Biochimica et Biophysica Academiae Scientiarum Hungaricae Acta Biologica et Medica Germanica Acta Chemica Scandinavica Acta Endocrinologica Advances in Clinical Chemistry Advances in Protein Chemistry Agricultural and Biological Chemistry American Journal of Opt halmo logy American Journal of Physiology Analytical Biochemistry Archives of Biochemistry and Biophysics Archiv fur die Gesamte Virusforschung Biochimica Biophysica Acta Biochimica Biophysica Acta Library Biochemical and Biophysical Research Communications The Biochemical Journal Biochemische Zeitschrift/European Journal of Biochemistry Biochemistry Biochemistry (Biokhimya) Biophysik Bulletin of the Agricultural Chemical Society of Japan Bulletin of the Japanese Society of Scientific Fisheries Bulletin de Ia Societe de Chimie Biologique Canadian Journal of Biochemistry Chemische Berichte/Berichte der Deutschen Chemischen Gesellschaft Clinica Chimica Acta Cold Spring Harbor Symposium on Quantitative Biology Collection Czechoslovak Chemical Communications Comparative Biochemistry and Physiology Comptes rendus Academie Science, Paris Comptes rendus des travaux du Laboratoire Carlsberg Endocrinology Enzymologia Experientia Experimental Eye Research Experimental Gerontology FEBS Letters Federation Proceedings Hoppe-Seyler's Zeitschrift fur Physiologische Chemie International Archives of Allergy and Applied Immunology International Journal of Protein Research International Journal of Radiation Biology Journal of Agricultural and Food Chemistry Journal of the American Chemical Society Journal of Bacteriology Journal of Biochemistry, Tokyo Journal of Biological Chemistry Journal of Biophysical and Biochemical Cytology Journal of Clinical Investigation Journal of Experimental Medicine Journal of General Virology Journal of Histochemistry and Cytochemistry Journal of Immunology Journal of Laboratory and Clinical Medicine Journal of Lipid Research Journal of Molecular Biology Journal of Neurochemistry Journal of the Optical Society of America Journal of Physiology Molecular Biology (Russian) Nature Proceedings of the Academy of Sciences, USSR (Doklady Akademii Nauk SSSR) Proceedings of the Koninklijke Nederlandse Akademie van Wetenschappen Proceedings of the National Academy of Sciences, USA Proceedings of the Royal Society of London Radiation Research Recueil des travaux chimiques des Pays-Bas Science Seminars in Hematology Thrombosis et Diathesis Haemorrhagica Virology Zeitschrift fur Naturforschung Books Biochemistry of the phagocytic process Localization and the role of myeloperoxidase and the mechanism of the halogenation process (Ed J Schultz) Amsterdam; North-Holland Publ Co., 1970 Biological and Chemical Aspects of Oxygenases (Eds K Bloch and O Hayaishi) Tokyo; Maruzen Co Ltd., 1966 C02: Chemical, Biochemical and Physiological Aspects (Eds R Forster, J Edsall, A Otis, and F Roughton) NASA SP-188 Washington, D C., 1969 The Haemostatic Mechanism in Man and Other Animals; the proceedings of a symposium (Ed R MacFarlane) New York-London; Academic Press, Inc., 1970 (Symposia of the Zoological Society of London, No 27) Metabolic Regulation and Enzyme Action FEES proceedings of the 6th meeting, Madrid (Eds A Sols and S Grisolia) New York; Academic Press, Inc., 1970 (FEES Symposia, Volume 19) Non-Heme Iron Proteins: Role in energy conversions (Ed A San Pietro) Yellow Springs, Ohio; Antioch Press, 1965 Prebiotic and Biochemical Evolution (Eds A Kimball and J Oro) New York, American Elsevier Pub Co., 1971 Probes of Structure and Function of Macromolecules and Membranes Volume 2: Probes of enzyme and hemoproteins (Ed B Chance) New York, Academic Press, Inc., 1971 !Proceedings of the 7th Congress European Society of Hematology Part II/2 New York-London; S Karger, 1959Pyridine NucIeotide-Dependent Dehydrogenases (Ed H Sund) New York; Springer Verlag, 1970 Specificity of Cell Surfaces (Eds B Davis and L Warren) New Jersey; PrenticeHall, Inc., 1967 The Svedberg (Eds A Tiselius and K Pedersen) Uppsala; Almquist and Wiksall Boktryckeri A B., 19Wj- I have gathered a bouquet of flowers from other men's gardens; naught but the string that binds them is my own Montaigne PREFACE The format of this volume is similar to that of Volume In this volume there are approximately 1150 spectra of proteins obtained under a variety of conditions This is twice the number of spectra found in Volume .For each figure, the source of the protein and the conditions under which the spectrum was taken are given if supplied by the original reference None of the spectrophotometric parameters so dear to spectroscopists are given as they were not given in the original references Unless otherwise stated, the spectra were taken of solutions in a 1-cm cell Approximately 8% of the references are to work published prior to 1960, 12% are for 1960-1965, 20/0 for 1966-1969, 27/o for 1970, 30/o for 1971, and 3$ for 1972; thus some 80/o of the references are to the literature covering the period from 1966 to early 1972 The spectra are arranged alphabetically and cross-indexed by source: animal, tissue, microbial, viral, plant and algae, yeast and fungi, and apoproteins There is also an index to those additives or modifiers which change the spectra of the proteins, e.g., a given protein and its nitro-derivative, or a given protein and the effect of a certain solvent upon its spectrum This volume and its predecessor should prove useful to anyone interested in protein spectra Specific examples of possible uses include the following: a protein chemist who desires to compare the spectrum taken in the 1950Ts with that of the spectrum of the same protein taken in the 1970*s; a hematologist who wishes to see the spectral differences among different hemoglobins; a researcher with the spectrum of a protein he has recently isolated who desires to compare some of its odd spectral features with spectra in these volumes; and a student who desires to know what kind of spectra proteins have in different solvents Standard abbreviations have been used in this volume and it has been assumed that readers know what they stand for The abbreviations used in the original references have been retained; thus DPN and DPNH, TPN and TPNH, NAD and NADH, and NADP and NADPH have been used, as well as AMP and ATP The Library of the Downstate Medical Center has been the major source of the necessary journals and books What they didnrt have available, they went out of their way to try and obtain for me During the summer months of 1971 and 1972 the Library of the Marine Biological Laboratory, Woods Hole, Massachusetts, was my "library-away-from-the-library." Here I was able to examine many of the nonmedical journals and books I should like to thank the librarians and staff of both libraries for their very necessary assistance I should also like to thank Mrs E Bakker for help in preparing the figures for publication; Miss M Colindreler for typing the indexes; Mrs E Levine for photocopying •work; and Mr W Kratil for photographing the figures If there are any errors I am responsible for them and I would appreciate having them brought to my attention Donald M Kirschenbaum Downstate Medical Center Brooklyn, New York 1973 Contents Acknowledgments vii Preface xii Protein Spectra: A ACE ACO ACY ADE ALB ALD 21 AMI 27 AMY 53 ANA 55 ANG 56 ARA 57 ARG 58 ARO 61 ASP 62 ATP 64 AZU 65 Protein Spectra: B 66 BAC 66 Protein Spectra: C 69 CAR 69 CAT 81 CEL 83 CER 84 CHL 85 CHR 87 This page has been reformatted by Knovel to provide easier navigation xiii xiv Contents CHY 90 COB 92 CON 93 CRY 95 CYC 98 CYS 99 CYT 104 Protein Spectra: D 172 DEO 172 DIM 173 DIP 174 DPN 175 Protein Spectra: E 176 ELA 176 ERA 177 ERY 179 Protein Spectra: F 181 FAC 181 FAT 182 FER 183 FIB 194 FLA 195 FRU 203 Protein Spectra: G 204 GAL 204 GEL 205 GLU 207 GLY 225 Protein Spectra: H 233 HAP 233 HEM 234 HEX 293 HIS 295 This page has been reformatted by Knovel to provide easier navigation 680 Index terms Links m,m-Dinitrophyenylsulfone Gelatin 206 Dioxane Cytochrome c 119 2,3-Diphosphoglycerate Hemoglobin 267 Dissociation Catalase 82 Dithiothreitol Lipoamide dehydrogenase 332 + DPN -pyruvate complex Lactic dehydrogenase 321 DPNH Alcohol dehydrogenase Lipoamide dehydrogenase 21 331 E Emasol Cytochrome oxidase 160 162 Epigallocatechin Polyphenol oxidase 444 1,2-Epoxy 3-(p-nitrophenoxy)-propane Pepsin 412 Ethoxyformyl Phosphofructokinase 432 Ethoxzolamide Carbonic anhydrase B 72 Carbonic anhydrase C 77 Carbonic anhydrase Co(II) 76 Ethylisocyanide Hemoglobin 248 N-EthyImaleimide Hemoglobin 246 N-Ethyl-5-phenylisooxazolium-3’-sulfonate Carboxypeptidase γval 80 This page has been reformatted by Knovel to provide easier navigation 253 681 Index terms Links F Ferric Ovotransferrin 407 Phycocyanin 439 Transferrin 537 Ferricyanide Hemoglobin 244 α-p-Mercuribenzoate hemoglobin chain 280 β-p-Mercuribenzoate hemoglobin chain 280 Protein, iron, succinylated 460 Sulfmyoglobin 379 248 261 250 261 Fluoride Hemoglobin 243 Nodules 398 Peroxidase 422 Formic acid - HCl Cytochrome C 118 Formyl Cytochrome C 119 Free radical Albumin 13 French press Membranes 358 Fructose Rat tongue extracts, l8o Fructose 1,6-diphosphate Aldolase 25 G Glucose Glucose oxidase 211 Ovalbumin 404 Rat tongue extract 180 Glutamate Glutamic acid dehydrogenase 219 Glutathione Glutathione reductase 224 This page has been reformatted by Knovel to provide easier navigation 212 264 682 Index terms Links Glycine Ribonuclease 500 Glyoxal Phosphorylase 436 Ribonuclease 508 Guanidine HCl Phosphorylase b 436 pyrophosphatase 479 p-Guanidinophenacyl Trypsin 542 H Heptenoate Glycolic acid oxidase 229 Homocysteinethiolactone + methylmercuric hydroxide Immunoglobin 313 Hydrogen Cytochrome 118 Hydrogen peroxide Hemolysates 289 Peroxidase 417 Peroxidase c 416 418 Sodium hydrosulfite Lipoyl dehydrogenase 108 344 3-Hydroxyanthranilic acid Albumin 16 erythro-β-Hydroxyaspartate Aminotransferase aspartic 48 glutamic-aspartic 49 49 Hydroxylamine Amine dehydrogenase 27 Glucose oxidase 211 Hydroxylamine + dithionite Cytochrome a 106 4-Hydroxy-6-mercaptopyrazolo (3,4-d)pyrimidine Xanthine oxidase 582 This page has been reformatted by Knovel to provide easier navigation 212 425 683 Index terms Links 2-Hyxiroxy-5-nitrobenzylbromide Carboxypeptidase A 78 Ribonuclease T1 501 Toxin 533 Hydroxypyridoxal phosphate Apoglutamic-oxaloacetic transaminase 536 Hydroxypyridoxamine phosphate Apoglutamic-oxaloacetic transaminase 536 Hydrazine sulfate Amine dehydrogenase 27 Hypoxanthine Xanthine oxidase 578 Illumination/Irradiation D-Amino and oxidase 35 Arainotransferase, aspartic 50 52 Ferredoxin 184 186 Flavodoxin 199 Glucose oxidase 214 NADH Dehydrogenase 388 NADPH-Sulfite reductase 391 Papain 410 Phage T2 427 Ribonuclease 511 Transhydrogenase 539 I Imidazole Tryptophanase 550 Indoleacetic acid Peroxidase a 419 Iodine Albumin 19 Arginine kinase 5-sulfenyl sulfonate 58 Diiodoangiotensin II 56 Erabutoxin 177 Glyceraldehyde 3-phosphate dehydrogenase 226 Insulin 313 This page has been reformatted by Knovel to provide easier navigation 420 684 Index terms Links Iodine (Continued) β-Lactoglobulin 10 Ovotransferrin 407 Peroxidase 48 Prealbumin 445 Retinol-binding protein 485 Iodoacetamide Lipoamide dehydrogenase 333 Iodoacetic acid Parathyroid hormone 411 Iproniazide Peroxidase 419 Isoleucine Aminotransferase, branched chain 44 K α-Ketoglutarate Aminotransferase, GABA 46 Aminotransferase, glutamic-aspartic 49 Imidazolylacetolphosphate-L-glutamate aminotransferase 310 L Lactate Cytochrome b2 109 Glycolic acid oxidase 227 Lactose β-Lactoglobulin 322 Lecithin Hemoglobin 266 Leucine L-Amino acid oxidase 40 Lipoic acid amide Lipoyl dehydrogenase 342 Lysine L-Lysine monooxygenase This page has been reformatted by Knovel to provide easier navigation 347 323 685 Index terms Links M Mercaptoethanol Cytochrome 104 Cytochrome complex 104 p-Mercuriphenyl sulfonic acid B-Phycoerythrin Mercury Coneanavalin 94 Methoxamide Carboxypeptidase γval 80 DL-α-Methylaspartate Aminotransferase, aspartic 47 Methylamine Amine dehydrogenase Methyl-modified α-Chymotrypsin 91 2-Methyl-1,4-naphthohydroquinone Peroxidase 422 N N-Methylnicotinamide Aldehyde oxidase 23 β-Lactalbumin 318 Lysozyme 353 + NAD Glyceraldehyde phosphate dehydrogenase 224 NADH Glutathione reductase 223 Lipoamide dehydrogeanse 332 Microsomes 365 Salicylate hydroxylase 517 NADH2 Lipoamide dehydrogenase 333 NADPH Glutathione reductase 223 Microsomes 365 Neoprontosil Carbonic anhydrase B 76 This page has been reformatted by Knovel to provide easier navigation 333 686 Index terms Links Neoprontosil (Continued) Carbonic anhydrase C 76 Nitric oxide Cytochrome cd 144 Cytochrome oxidase 158 NitroAlkaline phosphatase 429 Arginine kinase 58 Carbonic anhydrase, B 73 Carbonic anhydrase C 74 Cytochrome c 122 Glutamate dehydrogenase 217 Methemoglobin 270 Neurophysin II 394 Phosphatase 430 Ribonuclease 504 p-Nitrobenzene sulfonamide Carbonic anhydrase B, Co(II) 75 Carbonic anhydrase C, Co(II) 75 2-Nitro-4-carboxyphenyIsulfenyl Nuclease 401 Nitroguanyl Albumin 13 p-Nitrophenyl acetate Trypsin 541 2-Nitrophenylsulfenyl Glucagon 207 Lysozyme 351 Nuclease 401 Toxin 533 Nitroso Hemoglobin 327 Leghemoglobin 327 O L-Ornithine Arginine racemase 59 This page has been reformatted by Knovel to provide easier navigation 430 123 557 687 Index terms Links Oxalate Glycolic acid oxidase 229 Ozone Cobratoxin 92 Lysozyme 350 P Pepsin modified Albumin 17 Cytochrome c 123 cyanide complex 124 methylcarbylamine compound 124 nitrosobenzene compound 124 Globulin 207 Performic acid oxidized α-Chymotrypsin 90 Nuclease 400 Ribonuclease 501 Periodate Albumin 12 Globulin 206 Apocytochrome b-562 113 pH Carbonic anhydrase 78 Carbonic anhydrase B 75 77 Carbonic anhydrase C 74 75 Carbonic anhydrase, carboxymethyl 2+ 69 Carbonic anhydrase CO 74 Carboxypeptidase A 78 Concanavalin A 94 Cyanmethemoglobin A0 269 Cyanmethemoglobin A1 269 Cystathionine synthetase 99 Cytochrome c 122 Cytochrome c peroxidase 165 Cytochrome cc’ 134 Cytochrome c2 129 Cytochrome P-450 153 This page has been reformatted by Knovel to provide easier navigation 75 123 125 688 Index terms Links pH (Continued) Erabutoxin 177 Fibrinogen 194 Flagella 195 Flagellar protein 196 Foot-and-mouth disease virus, protein 565 Fructokinase 203 Galactothermin 204 Glucose oxidase 212 Glutamate dehydrogenase 218 Glycerol kinase 227 Glycolic acid oxidase 230 Glycoprotein 232 Glyoxylic acid reductase 232 Hamagglutinin 234 Haptoglobin 233 Hemocuprein 239 Hemoglobin 246 Hemoglobin, α-chains 281 Hemoglobin Toulouse 286 Keratinase 315 Laccase 317 Leghemiglobin 325 Leghemoglobin 328 Lipoprotein 339 Lipoxygenase 340 Lysozyme 351 α2-Macroglobulin 355 Myosin 385 Papain 409 Parvalbumin 411 Peroxidase 423 Phosphatase, nitrotyrosyl 429 Phospholipase A-I 434 Phospholipase A-II 434 Phosphorylase b 435 Protein alkylated 452 This page has been reformatted by Knovel to provide easier navigation 247 256 269 689 Index terms Links pH (Continued) fast component 465 haem 459 Hageman factor 460 myofibrullar 466 slow component 465 structural 449 469 Proteinase acid 448 neutral 448 Pyruvate kinase 482 Rhodopsin 489 Ribonuclease 496 Ribonuclease, performic acid oxidized 502 Ribonuclease T1 508 D-Serine dehydrase 518 Thioredoxin-II 528 Tobacco mosaic virus 572 Urocanase 555 o-Phenanthroline Glutamine PP-ribose-P-amidotransferase 219 Phenazine methosulfate Amine dehydrogenase 27 Fhenylalanine L-Amino acid oxidase 38 Phenylazobenzoy1 Amylase 54 Phenylglyoxal Ribonuclease T1 508 Phenylhydrazine Amine dehydrogenase 27 Protein, pink 456 Urocanase 556 Phenylpyruvate L-Amino acid oxidase 39 PhenyIthi ocarbamyl Insulin 314 This page has been reformatted by Knovel to provide easier navigation 503 690 Index terms Links Phospholipase Membranes 358 Phosphovitin Cytochrome c 125 Photo-oxidation Albumin Aminotransferase 42 Lysozyme 462 Proteins 462 Ribonuclease 495 Subtilopeptidase 523 Picolinimidylated Deoxyribonuclease 172 Pipecolate D-Amino acid oxidase 28 Piperidine 2-carboxylate D-Amino acid oxidase 28 Polyphenoloxidase Ribonuclease 501 Ribonuclease, performic acid oxidized 503 Potassium buffer Tyrosine phenol lyase 552 Potassium chloride RNA polymerase 512 Potassium nitrate Nitrate reductase 396 Potassium phosphate fhosphatase, alkaline 431 Pregnenolone complex Cytochrome P-450 152 Primaquine Hemoglobin 271 Protocatechuic acid Protocatechuate 3,4-dioxygenase Pyrocatechase 47 478 Pyridoxal 5’-phosphate Aminotransferase, aspartic This page has been reformatted by Knovel to provide easier navigation 49 691 Index terms Links Pyridoxal 5’-phosphate (Continued) Aminotransf erase, glutamic-aspartic 49 Cystathionine synthase 99 Hexokinase 293 Malate dehydrogenase 356 Tyrosine phenol lyase 552 Pyridoxal 5’-phosphate/sodium borohydride Hexokinase 293 Phosphoglucoisomerase 432 Pyridoxamine or pyridoxamine 5’-phosphate Aminotransferase aspartic 43 glutamic-aspartic 49 Arginine racemase 59 Imidazolylacetolphosphate-L-glutamate aminotransferase 311 Pyruvate D-Amino acid oxidase 30 Arginine racemase 59 PyruvaterFerredoxin oxido-reductase 480 ∆’-Pyrroline-2-carboxylate D-Amino acid reductase 29 R Reductively methylated Ribonuclease 503 S Salicylate L-Amino acid oxidase 39 Salicylate hydroxylase 517 Semicarbazide Amine dehydrogenase 27 Sodium buffer Tyrosine phenol lyase 552 Sodium chloride Tryptophanase 550 Sodium diethyldithocarbamate Ceruloplasmin 84 This page has been reformatted by Knovel to provide easier navigation 47 50 692 Index terms Links Sodium diethyldithocarbamate (Continued) Protein, pink 457 Sodium dodecyl sulfate Glyceraldehyde 3-phosphate dehydrogenase 225 Sodium phenobarbital D-Amino acid oxidase 32 Solvent effect Cytochrome c 114 119 Myoglobin 373 374 Sorbose Rat tongue extract 180 Subtilisin treated Resilin 44 Succinyl Aldolase 24 Ovotransferin 407 Phage 67 O-Succinyl-L-homoserine Cystathionine γ-synthase 101 O-Succinyl-L-serine Cystathionine γ-synthase 101 Sucrose Rat tongue extract l80 Sulfite Adenosine 5’-phosphosulfate reductase Bisulfite reductase 68 T Temperature affect Anacystis nidulans 55 Carboxypeptidase A 79 Catalase 81 Chlorella pyrenoidosa 85 Cystathionine synthase 99 Cytochrome b5 111 Cytochrome b-562 112 Cytochrome c 115 Erythrocuprein 180 This page has been reformatted by Knovel to provide easier navigation 120 121 693 Index terms Links Temperature affect (Continued) Glucose oxidase 212 Hemoglobin 243 Myoglobin 382 Peroxidase 166 Pigment, visual 442 413 416 413 260 264 Tetranitromethane Aminotransferase, aspartic Hemerythrin 50 236 Thioglycolate Catalase 81 5-Thio-2-nitrobenzoic acid Lipase LB 330 Time Albumin 12 D-Amino acid oxidase 33 Catalase 82 Ceruloplasmin 85 Cytochrome oxidase 163 Glucose oxidase 213 Glycolic acid oxidase 230 Hemoglobin 247 248 Peroxidase 416 423 Trypsin 541 Tetrafluorosuccinyl Phage 67 2,4,6-Trimethylpyrylium perchlorate Chymotrypsin 90 Trinitrophenyl Ribonuclease T1 510 Triphosphate of 6-mereapto-9-β-D-ribofuranosyl-purine 5’-phosphate Myosin Heavy subunit 384 Light subunit 383 Tryptophan L-Tryptophan 2,3-dioxygenase This page has been reformatted by Knovel to provide easier navigation 55 694 Index terms Links U Urea Calcium-binding protein 69 Cytochrome c3 131 Ferredoxin 191 Glutamine-PP-ribose-P amidotransferase 219 α-Lactalbumin 318 Lysozyme 353 Myoglobin 374 Retinol-binding protein 485 V Valerate D-Amino acid oxidase 35 W Woodwards Reagent K Carboxypeptidase αval 80 X Xanthine Xanthine oxidase 579 Z Zinc Aspartate transcarbamylase 62 regulatory subunit 62 Lipoamide dehydrogenase 336 This page has been reformatted by Knovel to provide easier navigation ... 0 .2 umole 3-lactoglobulin + 0 .2 umole Iodine in 2. 2 ml buffer Curve 2: 0 .2 umole albumin + 0 .2 umole iodine in 2. 2 ml buffer Curve 3: 0.6 umole iodine in 2. 8 ml water Curve k: 0.19 umole !3 in. .. microbial, viral, plant and algae, yeast and fungi, and apoproteins There is also an index to those additives or modifiers which change the spectra of the proteins, e.g., a given protein and its nitro-derivative,... include the following: a protein chemist who desires to compare the spectrum taken in the 1950Ts with that of the spectrum of the same protein taken in the 1970*s; a hematologist who wishes to see the