PROTIDES OF THE BIOLOGICAL FLUIDS

PROCEEDINGS OF THE TWENTY-EIGHTH COLLOQUIUM, 1980

Edited by

H. PEETERS L i p i d and P r o t e i n Department I n s t i t u t e for M e d i c a l Biology

Brüssels, Belgium

PERGAMON PRESS OXFORD • NEW YORK • TORONTO • PARIS • FRANKFURT • SYDNEY

U.K. Pergamon Press Ltd., Headington Hill Hall, Oxford OX3 OBW, England

U.S.A. Pergamon Press Inc., Maxwell House, Fairview Park, Elmsford, New York 10523, U.S.A. Pergamon of Canada, Suite 104, 150 Consumers Road, Willowdale, Ontar io M2J 1P9, Canada CANADA

AUSTRALIA Pergamon Press (Aust.) Pty. Ltd., P .O . Box 544, Pot ts Point , N .S .W. 2011, Australia

FRANCE Pergamon Press SARL, 24 rue des Ecoles, 75240 Paris , Cedex 05, France

FEDERAL REPUBL1C OF GERMANY

Pergamon Press G m b H , 6242 Kronberg-Taunus, Hammerweg 6, Federal Republic of Germany

Copyright© 1980 Pergamon Press Ltd. All Rights Reserved. No pari of thispublication may be reproduced, stored in a retrieval System or transmitted in any form or by any means: electronic, electrostatic, magnetic tape, mechanical, photocopying, recording or otherwise, without permission in writing from the publishers.

Library of Congress Catalog Card No. 58-5908

In order to make this volume available as economically and as rapidly as possible the authors' typescripts have been reproduced in their original forms. This method unfortunately has its typographical limitations but it is hoped that they in no way distract the reader.

Printcd in Great Britain by A. Wheaton & Co. Ltd., Exeter ISBN 0 08 0263704 ISSN 0079-7065

CONTENTS

PREFACE xix ACKNOWLEDGEMENTS xxi THE NINTH ARNE TISELIUS MEMORIAL LECTURE Plasma Cell Tumours of Man and Mouse 3

J. R. HOBBS

SECTION A. EVOLUTION OF PROTEINS A. l . Immunoglobulins Evolutionary Studies of Immunoglobulin Protein and Nucleic

Acid Sequences 17 W. C. BARKER and M. O. DAYHOFF

Structure, Function, and Evolution of Human Immunoglobulins 23 F. W. PUTNAM

Early Evolution of the Antibody Domain 29 A. D. McLACHLAN

Rabbit Antibody VL-Region Amino Acid Sequences Identification of J-Segments, an a Gene Interaction Model 33 D. G. BRAUN

v

vi CONTENTS

Characterisation of a Reptilian Macroglobulin Antibody J. D. WETHERALL and K. J. TURNER

A.2. Fibrinogen The Evolution of Vertebrate Fibrinogen

R. F. DOOLITTLE Fibrinogen Evolution: Intra- and Inter-species Comparisons

A. HENSCHEN, F. LOTTSPEICH, E. TÖPFER-PETERSEN, M. KEHL and R. TIMPL

Human Fibrinogen: Sequence, Sulfur Bridges, Glycosylation and Some Structural Variants A. HENSCHEN, F. LOTTSPEICH, C. SOUTHAN and E. TÖPFER-PETERSEN

A.3. Herne Proteins Mode, Tempo, and Role of Natural Selection in the Evolution of

Herne Proteins M. GOODMAN and J. CZELUSNIAK

On the Evolution of Bacterial Cytochromes C J. VAN BEEUMEN

Cytochromes C of Two Different Sequence Classes in Agrobacterium Tumefaciens J. VAN BEEUMEN, P. TEMPST, P. STEVENS, D. BRAL, J. VAN DAMME and J. DE LEY

Partial Amino Acid Sequence of Pig heart Left Ventricular Myosin Light Chain L27 X. GRANDIER-VAZEILLE, D. TETAERT, B. HEMON, K. K. HAN and G. BISERTE

Caiman Crocodylus Hemoglobin—Structure and Activity F. LECLERCQ, C. BAUER, A. FRABONI, C. PAUL, C. VANDECASSERIE, A. G. SCHNEK and G. BRAUNITZER

A.4. Metal Binding Protein Internal Duplication in the Structure of Human Ceruloplasmin

F. E. DWULET and F. W. PUTNAM

CONTENTS vii The Relationship of the Small Blue Proteins with the Copper-

Containing Oxidases 87 L. RYDEN and J.-O. LUNDGREN

Comparative Studies of Ferritins 91 M. HEUSTERSPREUTE, C. WUSTEFELD, J. M. MATHIJS, J. C. MARESCHAL, G. CHARLIER and R. R. CRICHTON

Calcium-binding Proteins: From Diversification of Structure to Diversification of Function 95 J. G. DEMAILLE, J. HAIECH and M. GOODMAN

A.5. Proteases Evolution of Haptoglobin and the Serine Proteases 99

A. KUROSKY Partial Sequence Comparison Between Human, Bovine and Porcine

Plasminogen 103 R. BRUNISHOLZ, P. MOSER, J. SCHALLER and E. RICKLI

Low Molecular Weight Protease 107 H.-J. TORFF, H. DÖRSAM and R. ZWILLING

Comparison of the Amino Acid Composition of Tryptic Peptides Between Calamary and Rabbit Muscle aldolase 111 J. C. MARESCHAL, J. WAUQUAIRE and R. R. CRICHTON

Crayfish Trypsin: Missing Link Between Procaryote and Mammalian Serine proteases 115 R. ZWILLING, H. NEURATH and R. G. WOODBURY

Mouse NGF y-Subunit and Crab Collagenase: Two Serine Proteases of Unusual Function 119 R. A. BRADSHAW, G. A. GRANT, K. A. THOMAS and A. Z. EISEN

A.6. Protease Inhibitors Evolution of the Family of Serine Proteinase Inhibitors Homologous to

Pancreatic Secretory Trypsin Inhibitor (Kazal) 123 M. LASKOWSKI, JR., I. KATO, W. J. KOHR, C. J. MARCH and W. C. BOGARD

Homology of er-Antitrypsin and Antithrombin-III Identification of Active Sites 129 D. R. BOSWELL, M. C. OWEN, S. O. BRENNAN, R. W. CARRELL and A.D. McLACHLAN

viii CONTENTS

A.7. Ribonuclease The Molecular Evolution of Mammalian Pancreatic Ribonuclease 133

J. J. BEINTEMA, J. A. LENSTRA and W. M. FITCH Ribonuclease Variation Within the Species 139

J. J. BEINTEMA

A,8. Various Proteins Complete Primary Structure of the Hemagglutinin from Two Different Strains of

Influenza A Type Virus 143 W. MIN JOU, M. VERHOEYEN, R.-X. FANG, R. DEVOS, D. HUYLEBROECK, E. SAMAN and W. FIERS

a-Lactalbumin: Studies of the Binding Site for Galactosyltransferase 149 K. BREW and S. K. SINHA

Evolutionary Diversity in the Structure and Activity of Carbonic Anhydrase 153 R. E. TASHIAN, D. HEWETT-EMMETT and M. GOODMAN

Mutations Affecting the Structure of the Alcohol Dehydrogenase from Drosophila Melanogaster 157 D. R. THATCHER and A. RETZIOS

Trends in the Molecular Evolution of a-Crystallin 161 W. W. DE JONG, A. ZWEERS and M. GOODMAN

Insulin-related Growth Factors 165 R. A. BRADSHAW

Pseudo-oligomer Theory to Explain Evolution of Soluble Proteins and their Binding to Receptors 169 R. GRÄSBECK and I. KOUVONEN

On the Origin and Relation of Human Seminal Plasma Proteins to Plasma Proteins 173 J. LIZANA, P. ENEROTH and M. BYGDEMAN

Identification of Reptilian Complement Component C3 J. D. WETHERALL, G. SIMMS and N. GUDE 179

CONTENTS ix

Comparative Molecular Aspects of the Chicken Major Histocompatibility Complex Coded Alloantigens 183 C.-H. BROGREN and S. BISATI

SECTIONB. MOLECULAR BIOLOGY OF BLOOD COAGULATION AND FIBRINOLYSIS B.l . Contact Activation The Role of Factor XII (Hageman Factor) in Blood Coagulation and a Partial

Amino Acid Sequence of Human Factor XII and its Fragments 193 K. FUJIKAWA, B. McMULLEN, R. L. HEIMARK, K. KURACHI and E. W. DAVIE

Mechanism of Activation of Human Coagulation Factor XI and Plasma Prekallikrein 197 B. N. BOUMA and J. H. GRIFFIN

The Hageman Factor-Kallikrein System in Inflammatory Injury 201 C. G. COCHRANE

Functions of the Kallikrein-Kinin System Outside the Clotting Cascade 205 G. L. HABERLAND

The Ellagic Acid-Dependent Activation of Factor XI by Factor XII 209 J. S. GREENGARD, G. N. BERETTA and J. H. GRIFFIN

Structural and Functional Aspects of Human High-molecular Weight (HMW) Kininogen 213 B. DITTMANN, F. LOTTSPEICH, A. HENSCHEN and H. FRITZ

Recent Studies of Human High Molecular Weight Kininogen 217 D. M. KERBIRIOU, B. N. BOUMA and J. H. GRIFFIN

Intrinsic Kallikrein- and Plasmin-Dependent Pathways of Activation of the Renin-angiotensin System in Plasma 221 F. DERKX, B. BOUMA, J. DE BRUYN and M. SCHALEKAMP

Dextran Sulfate Induced Activation of Prekallikrein in Normal Human Plasma 225 F. VAN DER GRAAF, J. F. A. KEUS, R. A. A. VLOOSWIJK and B. N. BOUMA

X CONTENTS Efficacy of Combined Treatment of Factor IX-Complex (PPSB) with

/3-Propiolactone (ß-PL) and Ultraviolet (UV) Irradiation 229 W. STEPHAN and A. M. PRINCE

The Intrinsic and Extrinsic Activation of Factor IX 233 B. <{)STERUD

B.2. Extrinsic System The Tissue Factor Pathway of Coagulation: A Preliminary Kinetic Model 237

Y. NEMERSON, R. GENTRY, M. ZUR and R. BACH Tissue Thromboplastin—Molecular and Cellular Biology 241

H. PRYDZ and T. LYBERG Factor VII Activation in Plasma in the Presence of Activated Platelets 245

B. 4)STERUD

B.3. Prothrombin The Carboxylation of Prothrombin Precursors in a Bovine Liver Cell-free System 249

C. VERMEER, M. DE METZ, B. A. M. SOUTE and H. C. HEMKER Properties of the Vitamin K-Dependent Carboxylase 253

J. W. SUTTIE, J. McTIGUE and A. E. LARSON X-Ray Studies of Fragment 1 of Bovine Prothrombin 257

C. C. F. BLAKE, J. M. BURRIDGE and S. J. OATLEY High Resolution Nuclear Magnetic Resonance Spectroscopy of Bovine

Prothrombin Fragments 261 M. P. ESNOUF, E. A. ISRAEL, N. D. PLUCK and R. J. P. WILLIAMS

On the Role of Phospholipids in the Reaction Mechanism of Blood Coagulation 265 H. C. HEMKER, G. VAN DIEIJEN, J. RÖSING, G. TANS and R. F. A. ZW AAL

The Platelet Release Reaction and the Activation of Prothrombin 273 H. SANDBERG and L. -O. ANDERSSON

Dynamic Interaction of Prothrombin and Fragment I with Phospholipid Monolayers 277 M. F. LECOMPTE, J. ELION and I. R. MILLER

Effect of Anticoagulant Phospholipase A2 on Prothrombin Activation M. C. BOFFA, M. C. BRYCKAERT and M. P. ESNOUF 281

CONTENTS Prothrombin and Stored Oxalated Plasma

C. GRAY and D. PERRY

B.4. Factor VIII The Properties of Human Factor VIII Procoagulant Protein

L. W. HOYER and N. C. TRABOLD Anomalous Behaviour of Factor VIH-Von Willebrand Factor Complex as

a Substrate for Serine Proteases J. A. VAN MOURIK, P. H. G. LANTINGA and J. A. HELLINGS

Chemical Composition of Factor VIII C. MAZURIER, B. SAMOR, A. PARQUET-GERNEZ, B. FOURNET, M. GOUDEMAND and J. MONTREUIL

Binding to Gold Granules Enhances Ristocetin Cofactor Activity of Low Molecular Weight Factor VIII M. FURLAN, B. A. PERRET and E. A. BECK

B.5. Fibrinogen and Fibrin Paracrystalline Structure of Native Human Fibrinogen

R. GOLLWITZER, W. BODE, H. J. SCHRAMM, D. TYPKE and R. GUCKENBERGER

Synthetic Peptide Probes and the Location of Fibrin Polymerization Sites R. F. DOOLITTLE and A. P. LAUDANO

Structure of Fibrin Oriented in Strong Magnetic Fields G. HUDRY-CLERGEON, J. M. FREYSSINET and J. TORBET

Calcium and the Structure of Fibrin(ogen) Fragments D F. HA VERKATE and W. NIEUWENHUIZEN

A Light Scattering Study of the Kinetics of Fibrin Polymerisation A. VISSER and T. A. J. PA YENS

Fibrin/Fibrinogen Degradation Products (FDP) Analysis Using Laser Nephelometry R. SUENAGA, Y. HAMAMURA, N. YOSHIOKA, M. AGETA and S. YAMASAKI

Fibrinogen—Coaguable or Inhibitor—by Treatment with /J-Propiolactone R. KOTITSCHKE and W. STEPHAN

xii CONTENTS

B.6. Inhibitor of the Clotting System A Human Inhibitor of Hageman Factor Acting as a Fibrinolytic Inhibitor 337

U. HEDNER Anticoagulant Action of Human Protein C 341

R. A. MARLAR, A. J. KLEISS and J. H. GRIFFIN Discrepancies Between Antigen Concentrations and Functional

Activities of Plasma Protease Inhibitors 345 M. J. GALLIMORE

Characterization of Antithrombin III Deficiency in a Japanese Family 349 O. MATSUO, Y. OHKI, S. KONDO and T. MATSUO

Antithrombin III Microheterogeneity Demonstrated by Agarose Isoelectric Focusing 353 A. MILNER, D. BURNETT, R. DREW, D. RAMSDEN and A. R. BRADWELL

Proteinase Inhibitors, Proteinases, Pro-Arylamidase in Fertilized and Unfertilized Eggs of Salmonids 357 U. C. VIETH and G. HERMANN

B.7. Fibrinolytic System Regulation and Control of Fibrinolysis 361

D. COLLEN Fibrin-binding of Native Fragments of Known Primary Structure from

Human Plasminogen 365 S. THORSEN, I. CLEMMENSEN, L. SOTTRUP-JENSEN and S. MAGNUSSON

Investigations on the Identification and Localization of Lysine Binding Sites of Human Plasminogen 371 P. G. LERCH and E. E. RICKLI

Plasminogen Proactivators and Activators in Human Plasma: Factor XII-Dependent Fibrinolysis 375 C. KLUFT

On the Reaction Between Plasmin and a2-Antiplasmin 379 B. WIMAN, T. NILSSON and D. COLLEN

CONTENTS xiii A Novel Method for the Preparation of Plasmin enabling the Study of Glu-Plasmin

and its Role in Fibrinolysis 383 E. SUENSON, O. LÜTZEN and S. THORSEN

Human Tissue Plasminogen Activator and its Occurrence in Blood 387 D. C. RIJKEN and G. WIJNGAARDS

The Effect of Fibrin on the Activity of Purified Human Vascular Plasminogen Activator 391 B. R. BINDER and J. SPRAGG

Inhibition of Plasmin or Plasmin-Streptokinase Complex by Aprotinin or a2-Antiplasmin 395 B. WIMAN

Maintenance of the Aprotinin-Binding Domain in Plasmin-Derived Light Chain as Shown by Affinity Chromatography 399 J. CHAUVET and R. ACHER

Comparison of Fibrinolytic and Amidolytic Measurements of Urokinase Activity in Urine 405 D. PAAR and D. MARUHN

Relationship Between Activity Losses and the Conversion of High to Low Molecular Weight Urokinase 409 K. HUBER and B. R. BINDER

Effect of Dextran Sulphate on the S-2251 Splitting Activity of Human Plasma 413 O. MATSUO and H. R. LÜNEN

Clotting and Other Plasma Factors in Septicemia: Inhibition of Degradation by an Exogenous Protease Inhibitor from Soybeans (Type: Bowman-Birk) 417 M. JOCHUM, J. WITTE, H. SCHIESSLER and H. FRITZ

Unusual Kinetic Behaviour of Plasmin (Fibrinolysin) Digestion of Fibrin and N-Carbobenzoxy-Tyrosine-Nitrophenyl-Ester 421 F. S. STEVEN, S. QAZZAZ, M. M. GRIFFIN and B. S. BROWN

Fibrinolysis in Human Plasma S. BARLATI and M. COLOMBI 427

xiv CONTENTS

SECTION C. HYBRIDOMAS AND MONOCLONAL ANTIBODIES C.l. Production of Monoclonal Antibodies Secretion of Human Immunoglobulins in Man-Mouse Hybridomas:

Correlation with 02-Microglobulin and Chromosome IS 433 H. JACOT-GUILLARMOD, G. STAIGER and V. MIGGIANO

Immunoglobulin M Mutants 439 G. KÖHLER and M. SHULMAN

Enhancement of the Growth of Hybridomas by Human Endothelial Culture Supernatant (HECS) 443 G. C. B. ASTALDI, M. C. JANSSEN, P. LANSDORP, W. P. ZEIJLEMAKER and C. WILLEMS

Murine Cytolytic T-Lymphocyte Hybrids 447 A. CONZELMANN, M. CIANFRIGLIA, O. ACUTO, W. HAAS, H. V. BOEHMER, H. POHLIT, J. P. JOHNSON and M. NABHOLZ

T Cell Hybrids Producing Antigen Specific Factors 451 S. KONTIAINEN, J. M. CECKA, E. CULBERT, E. SIMPSON and M. FELDMANN

Production of Anti-Streptococcal Group A Antibodies by Hybridization Following in vitro Immunization 459 H. HERBST and D. G. BRAUN

HLA Antigens Temperature Dependent Elution from Specific Monoclonal Antibody Columns 463 G. GAROTTA and M. TRUCCO

Binding Kinetics of Monoclonal Antibodies to Cell Surface Antigens 467 D. W. MASON and A. F. WILLIAMS

Competition and Affinity Assay of Monoclonal Antibodies Against Human Growth Hormone 471 J. IVANYI

Detection of Monoclonal Antibodies Against Cell Surface Antigens Using Immunoperoxidase Procedures 475 P. M. LANSDORP, G. C. B. ASTALDI, F. OOSTERHOF and W. P. ZEIJLEMAKER

CONTENTS xv Attachment of Lymphoid Cells to Glutaraldehyde Activated Plastic Cups:

A Convenient Means for Quantitative Estimation of Cytoplasmic and Membrane Ig 479 M. STANISLAWSKI, A. HAREL, M. MITARD and J. PENE

Purification of IgE Heavy Chain mRNA from a Monoclonal Mouse Hybridoma 483 M. ALBRING and I. BÖTTCHER

C.2. Monoclonal Antibody Conformation Modification of Protein Conformation Induced by Monoclonal

Antibodies: The ß Galactosidase Model 489 R. S. ACCOLLA, R. CINA, E. MONTESORO and F. CELADA

Structural Analysis of Hybridoma Derived Anti-p-Azobenzene-Arsonate (ABA) Antibodies 493 S. S. ALKAN and D. G. BRAUN

C.3. Monoclonal Antibodies and Cancer Conjugates Between a Toxin Subunit and Monoclonal Antibodies

(Immunotoxins) with High Specific Cytotoxicity 497 B. PAU, H. BLYTHMAN, P. CASELLAS, O. GROS, P. GROS, F. K. JANSEN, F. PAULUCCI, H. VIDAL and G. A. VOISIN

Monoclonal Antibodies to Three Tumor Markers: Human Chorionic Gonadotropin (hCG), Prostatic Acid Phosphatase (PAP) and Carcinoembryonic Antigen (CEA) 501 V. MIGGIANO, C. STÄHLI, P. HÄRING, J. SCHMIDT, M. LE DAIN, B. GLATTHAAR and T. STAEHELIN

Demonstration of Human Melanoma Associated Antigen(s) by Monoclonal Antibodies 505 S. CARREL, R. S. ACCOLLA, A. L. CARMAGNOLA and J.-P. MACH

Use of Monoclonal Anti-CEA Antibodies in Immunoadsorbent Columns and Solid-Phase Radioimmunoassay 511 F. BUCHEGGER, R. S. ACCOLLA, S. CARREL, A. CARMAGNOLA, C. GIRARDET and J.-P. MACH

Monoclonal Antibodies Against Carcinoembryonic Antigen (CEA) G. T. ROGERS, G. A. RAWLINS and K. D. BAGSHAWE 517

xvi CONTENTS Standardization of Monoclonal Antibodies for Clinical Studies in

Leukemia/Lymphoma 523 G. JANOSSY, N. TIDMAN, D. CRAWFORD, E. S. PAPAGEORGIOU, H. G. PRENTICE, G. FRANCIS, K. F. BRADSTOCK, I. McCONNELL, D. SECHER and C. MILSTEIN

C.4. Monoclonal Antibodies Against Cells and Lymphocytes Antigen-Specific Suppressor Factor from a T-Hybrid Line and its B

Cell Acceptor 529 M. J. TAUSSIG

Mouse Monoclonal Antibodies Raised Against Human T Lymphocytes and Melanoma Cells 533 J. JOHNSON, T. MEO, M. HADAM and G. RIETHMÜLLER

Monoclonal Antibody Specific for Normal and Neoplastic Human T Cells 537 I. ROYSTON, J. A. MAJDA, G. Y. YAMAMOTO and S. M. BAIRD

Normal Values of Human Lymphocyte Subsets Defined by Monoclonal Antibodies 541 B. VAN CAMP, M. DE WAELE and A. CRIEL

T Cell Hybridomas Mediating Specific Helper Cell and Cytotoxic Cell Activities 545 Z. ESHHAR, R. N. APTE, E. MOZES, T. WAKS, I. LÖWY, G. BERKE and Y. KAUFMANN

Inhibition of Mixed Lymphocyte Reaction by a Monoclonal Antibody Specific for a Rat T Cell Subset 549 M. WEBB

Monoclonal Antibodies to Cell Surface Antigens on Cells of the Mouse Lymphoid System. Histophysiological Applications 553 P. W. VAN VLIET, W. VAN EWIJK, J. ROZING and P. L. VAN SOEST

C.5. Monoclonal Antibodies Against Molecüles The Repertoire of Anti-Influenza Antibodies 559

S. FAZEKAS DE ST. GROTH, P. A. UNDERWOOD and D. SCHEIDEGGER The Silent Repertoire: Antibodies Produced in Mice Suppressed for

a Dominant Idiotype 565 C. BEREK, H. P. KOCHER, M. H. SCHREIER, J.-C. JATON and H. COSENZA

CONTENTS xvii Monoclonal Anti-Idiotypes as a Probe for the Analysis of the Diversity

of Anti-Phosphorylchoüne Antibodies 569 G. J. HÄMMERLING and R. WALLICH

Characterization of a Syngenic Monoclonal Anti-Idiotypic Antibody to a Monoclonal Anti-Beef APO-Cytochrome c Antibody Obtained by Somatic Cell Fusion 575 G. CORRADIN and R. ACCOLLA

Antigenic Variation of H-2 Molecules 579 D. GÖTZE, L. HOFMANN, H.-C. HUANG and H.-P. VOLLMERS

Fine Antigenic Specificity of RNA-Binding Monoclonal Antibody from Autoimmune NZB/NZW Mouse 583 D. EILAT, S. A. BEN-SASSON and R. LASKOV

Production of Monoclonal Antibodies Against HLA-DR Antigens and the Human Plasma Plasminogen-Activator 587 J. KALIL, E. ANGLES-CANO, D. CREVAT, Y. SULTAN and M. FELLOUS

Subsets of Human Ia-like Molecules Defined by Monoclonal Antibodies 595 N. GROSS, S. CARREL, R. TOSI, N. TANIGAKI, A. CARMAGNOLA and R. S. ACCOLLA

Serological and Functional Characterization of Monoclonal Antibodies to Guinea Pig Ia-Antigens 599 R. BURGER and E. M. SHEVACH

Monoclonal Antibodies as Tools for the Study of Idiotype-Antiidiotype Interactions Along the Anti-Levan Immunization Chain 603 P. LEGRAIN, D. VOHGTLE, P. A. CAZENAVE and G. BUTTIN

Monoclonal Antibodies to Human IgG; Finding of a Hybridoma Producing two Classes of Immunoglobulin (IgM and IgG) with Specificity for the same Antigen 607 G. GUARNOTTA, F. C. HAY, I. M. ROITT and M. G. JONES

SUBJECT INDEX 611

B. 7. Fibrinolytic System CLOTTING AND OTHER PLASMA

FACTORS IN SEPTICEMIA: INHIBITION OF DEGRADATION BY

AN EXOGENOUS PROTEASE INHIBITOR FROM SOYBEANS

(TYPE: BOWMAN-BIRK) M. JOCHUM, J. WITTE, H. SCHIESSLER and H. FRITZ

Department of Clinical Chemistry and Biochemistry, Surgical Ciinic University of Munich, Federai Repubiic of Germany

ABSTRACT

In the course of Gram-negative s e p t i c e m i a , s e p t i c shock, or e x p e r i -mental endotoxemia g r a n u l o c y t i c p r o t e i n a s e s such as e l a s t a s e and c a -t h e p s i n G are suggested to be r e l e a s e d by S t i m u l a t i o n with endotoxin. In v i t r o p r o t e o l y s i s of s e v e r a l plasma f a c t o r s by g r a n u l o c y t i c e l a ­s t a s e i s almost completely prevented by p r i o r i n h i b i t i o n of e l a s t a s e with a P r o t e i n a s e i n h i b i t o r from soybeans (Bowman-Birk i n h i b i t o r , B B I ) . In experimental endotoxemia i n dogs, degradation o f v a r i o u s plasma f a c t o r s c o u l d a l s o be s i g n i f i c a n t l y d i m i n i s h e d by p r i o r systemic ap-p l i c a t i o n o f t h i s e l a s t a s e - c a t h e p s i n G i n h i b i t o r (BBI). These r e s u l t s i n d i c a t e t h a t the consumption r e a c t i o n observed d u r i n g endotoxemia and se p t i c e m i a i s a b i p h a s i c event. Besides a c t i v a t i o n of the c l o t t i n g and f i b r i n o l y t i c Systems by s y s t e m - s p e c i f i c p r o t e i n a s e s such as throm-bokinase or Plasminogen a c t i v a t o r , the decrease of the l e v e l s of p l a s ­ma f a c t o r s i s caused a l s o to an a p p r e c i a b l e degree by u n s p e c i f i c p ro-t e o l y t i c degradation due to g r a n u l o c y t i c p r o t e i n a s e s .

KEYWORDS

Septicemia; endotoxemia; endotoxin; disseminated i n t r a v a s c u l a r coagu­l a t i o n (DIC); u n s p e c i f i c p r o t e o l y s i s ; g r a n u l o c y t i c p r o t e i n a s e s ; Pro­t e i n a s e i n h i b i t o r .

INTRODUCTION

Human polymorphonuclear (PMN) leu k o c y t e s c o n t a i n a t l e a s t three neu­t r a l p r o t e i n a s e s ( e l a s t a s e , c a t h e p s i n G, collagenase) which are r e ­le a s e d by S t i m u l a t i o n with endotoxin. T h i s c o u l d be p a r t i c u l a r l y de-monstrated f o r g r a n u l o c y t i c e l a s t a s e i n the course of acute myeloic leukemia and septicemia (Egbring and co-workers, 1977) as w e l l as i n canine endotoxin shock. (Aasen and Ohlsson, 1978). The a c t i v i t i e s of c l o t t i n g and other plasma f a c t o r s e i t h e r i n t h e i r i s o l a t e d form or i n plasma are s t r o n g l y reduced by d i g e s t i o n with e l a s t a s e or c a t h e p s i n G (Johnson and co-workers, 1976; Schmidt and co-workers, 1974).

4 1 7

4 1 8 FIBRINOLYTIC SYSTEM

Even i n v i v o the r e d u c t i o n of such f a c t o r s could be due to degrada­t i o n by g r a n u l o c y t i c p r o t e i n a s e s under c e r t a i n p a t h o l o g i c a l c o n d i -t i o n s (Egbring and Havemann, 1978). The a c t i v i t y o f the g r a n u l o c y t i c p r o t e i n a s e s i s r e g u l a t e d i n v i v o by plasma i n h i b i t o r s such as oC* - a n t i t r y p s i n , öC 2-macroglobulin and eC-a n t i c h y m o t r y p s i n and by a n t i l e u k o p r o t e a s e i n mucous s e c r e t i o n s (ror r e f e r e n c e s see Aasen and Ohlsson, 1978). The i n v i t r o i n t e r a c t i o n of g r a n u l o c y t i c e l a s t a s e and c a t h e p s i n G with i n h i b i t o r s o f animal, p l a n t and m i c r o b i a l o r i g i n i s w e l l i n v e s t i g a t e d ( S c h i e s s l e r and co-workers, 1977). Only r e c e n t l y , i n h i b i t i o n of g r a n u l o c y t i c p r o t e i n a s e s c o u l d be demonstrated i n d i r e c t l y a f t e r a p p l i c a t i o n of an e l a s t a s e - c a t h e p s i n G i n h i b i t o r from soybeans (BBI; MW 8000) i n the course of experimental Gram-negative septicemia i n dogs ( S c h i e s s l e r and co-workers, 1978). As i n f u s i o n of BBI prevented the r a p i d f a l l i n a c t i v i t y o f the f i b -r i n - s t a b i l i z i n g f a c t o r F X I I I , the authors suggested t h a t t h i s de-crease might be a l s o caused by n o n s p e c i f i c p r o t e o l y s i s r a t h e r than by the a c t i o n of thrombin throughout severe disseminated i n t r a v a s c u l a r c o a g u l a t i o n (DIC) which o f t e n complicates the p a t h o p h y s i o l o g i c a l events i n s e p t i c e m i a . The purpose of t h i s i n v e s t i g a t i o n was to show th a t the decrease i n c l o t t i n g and other plasma f a c t o r s d u r i n g s e p t i ­cemia, s e p t i c shock, or experimental endotoxemia i s not o n l y due to DIC (Müller-Berghaus and Lasch, 1975) but a l s o to an a p p r e c i a b l e de-gree to u n s p e c i f i c p r o t e o l y s i s by g r a n u l o c y t i c p r o t e i n a s e s .

RESULTS

In v i t r o , p u r i f i e d human g r a n u l o c y t i c e l a s t a s e degraded i s o l a t e d c l o t ­t i n g f a c t o r s (F V I I , F Xa, F XIII) and the t r a n s p o r t p r o t e i n t r a n s -f e r r i n r a p i d l y w i t h i n 5-30 minutes even at a molar r a t i o of 2 ( e l a ­stase) to 100 (plasma p r o t e i n ) . T h i s c o u l d be demonstrated by PAG-e l e c t r o p h o r e s i s and immunoelectrophoresis according to L a u r e l l . Ad-d i t i o n a l l y , the major i n h i b i t o r of c l o t t i n g f a c t o r s a n t i t h r o m b i n I I I i s a l s o r a p i d l y degraded by e l a s t a s e thereby l o o s i n g i t s b i o l o g i c a l a c t i v i t y . T h i s was shown by c r o s s e d immunoelectrophoresis as w e l l as with an a c t i v i t y assay using the chromogenic Substrate S-2238 from DeutscheKabi, München (for d e t a i l e d Information: M. Jochum, H. Lan­der, and H. F r i t z , manuscript i n p r e p a r a t i o n ) . P r o t e o l y t i c degrada­t i o n and l o s s of a c t i v i t y was almost completely i n h i b i t e d by p r i o r i n c u b a t i o n of e l a s t a s e with BBI. The plasma l e v e l s of antithrombin I I I and «sC-macroglobulin as w e l l as those of v a r i o u s c l o t t i n g ( f i b r i n o g e n , F X I I I ) , complement (C3, C4) and other plasma f a c t o r s ( t r a n s f e r r i n , prealbumin, inter-OC-trypsin i n h i b i t o r ) were found to be s i g n i f i c a n t l y decreased i n 18 p a t i e n t s s u f f e r i n g from hyperdynamic s e p t i c shock (experimental d e t a i l s are g i v e n i n the h a b i l i t a t i o n S c r i p t of Witte, 1979 and are submitted f o r p u b l i c a t i o n ) . A s i m i l a r s t a t i s t i c a l l y s i g n i f i c a n t r e d u c t i o n of the c o n c e n t r a t i o n s and a c t i v i t i e s of s e v e r a l plasma p r o t e i n s (Pro­thrombin, antithrombin I I I , f a c t o r F X I I I , Plasminogen, d^-plasmin i n h i b i t o r , and complement f a c t o r C3) was observed i n experimental endotoxemia i n dogs (M. Jochum, J . Witte, H. S c h i e s s l e r , G. Ruck-de s c h e l , and H. F r i t z , manuscript i n p r e p a r a t i o n ) . In t h i s model, the decrease of the plasma p r o t e i n s was c o n s i d e r a b l e d i m i n i s h e d by p r i o r intravenous i n j e c t i o n of the e l a s t a s e - c a t h e p s i n G i n h i b i t o r BBI ( F i g . 1).

CLOTTING AND OTHER PLASMA FACTORS IN SEPTICEMIA 4 J 9

F i g . 1 Plasma l e v e l s of s e v e r a l plasma p r o t e i n s during experimental endotoxemia without (a) and with (b) Bowman-Birk i n h i b i t o r medi-c a t i o n . At the Ordinate the percentage of the s t a r t i n g plasma l e v e l of each f a c t o r i s given. At the a b s c i s s a the experimental p e r i o d and the endotoxin i n f u s i o n p e r i o d ( t h i c k l i n e ) are i n d i c a t e d . For f u r t h e r d e t a i l s see Witte, 1979.

DISCUSSION

Marked consumption or turnover of plasma P r o t e i n a s e i n h i b i t o r s during s e p t i c shock r e f l e c t extensive a c t i v a t i o n of the c l o t t i n g , f i b r i n o ­l y t i c , and complement Systems (summarized as "blood Systems") as w e l l as the l i b e r a t i o n of c o n s i d e r a b l e amounts of p r o t e i n a s e s from v a r i o u s body c e l l s , e s p e c i a l l y from g r a n u l o c y t e s ( f o r r e f e r e n c e s see F r i t z , 1980). S y s t e m - s p e c i f i c p r o t e i n a s e s such as Plasminogen a c t i v a t o r and thrombokinase a c t i v a t e the blood Systems by s p e c i f i c p r o t e o l y t i c c l e a v a g e s , i . e . by proenzyme-» enzyme c o n v e r s i o n . l e a d i n g to severe consumption of plasma p r o t e i n s . The r e l e a s e of g r a n u l o c y t i c p r o t e i n ­ases causes furthermore c o n s i d e r a b l e u n s p e c i f i c d egradation of the plasma f a c t o r s as c o u l d be demonstrated i n d i r e c t l y by systemic a p p l i -c a t i o n of an e l a s t a s e - c a t h e p s i n G i n h i b i t o r (BBI) i n the course of experimental endotoxemia. However, g u a n t i t a t i o n of the degree of con­sumption due to u n s p e c i f i c p r o t e o l y s i s i s not f e a s i b l e y e t . The r e s u l t s of our experimental and c l i n i c a l s t u d i e s i n d i c a t e c l e a r l y t h a t i n ge-n e r a l i z e d inflammatory processes l i k e s e p t i c e m i a or s e p t i c shock the

4 2 0 FIBRINOLYTIC SYSTEM

r e g u l a t o r y i n h i b i t o r p o t e n t i a l of the organism may be o v e r s t r e s s e d . Systemic a p p l i c a t i o n of s u i t a b l e P r o t e i n a s e i n h i b i t o r s may prevent a t l e a s t the u n s p e c i f i c degradation of plasma f a c t o r s and t h e r e f o r e c o n t r i b u t e to m a i n t a i n the p h y s i o l o g i c a l balance. As medication w i t h BBI or other P r o t e i n a s e i n h i b i t o r s i s not p o s s i b l e i n humans u n t i l t o x i c o l o g i c a l , pharmacological and c l i n i c a l t r i a l s have been success-f u l l y e l a b o r a t e d , 'Substitution with a n t i t h r o m b i n I I I and o^-macroglo-b u l i n i s the method of c h o i c e a t present.

AC KNOWLEDGEMENT

T h i s work has been supported by grants from the"Sonderforschungsbe­r e i c h 51 - Munich. We are g r a t e f u l l y indebted to a l l c o l l e a g u e s and t e c h n i c a l a s s i s t a n t s who took g r e a t p a r t i n t h i s study.

REFERENCES

Aasen, A. I., and K. Ohlsson (1978). Hoppe-Seyler 1s Z. P h y s i o l . Chem., 359, 683-690.

Egbring, R., W. Schmidt, G. Fuchs, and K. Havemann (1977). Blood, 49, 219-231.

Egbring, R., and K. Havemann (1978). In: Havemann, K., and A. J a n o f f (Eds.), N e u t r a l Proteases of Human Polymorphonuclear Leukocytes. Urban and Schwarzenberg, Inc. Baltimore-Munich, pp. 44 2-4 58.

F r i t z , H. (1980). In: Evered, D., and J . Whelan (Eds.), P r o t e i n De­g r a d a t i o n i n Health and Disease, C i b a Foundation Symposium 75, Ex-c e r p t a Medica, Amsterdam.

Johnson, U., K. Ohlsson, and I. Olsson (1976). Scand. J . Immunol., 5, 421-426.

Müller-Berghaus, G., and H. G. Lasch (1975). In: Urbaschek, B., R. Urbaschek, and E. Neter (Eds.), Gram-Negative B a c t e r i a l I n f e c t i o n s and Mode of Endotoxin A c t i o n s . Springer, Wien - New York, pp. 353-356.

S c h i e s s l e r , H., K. Ohlsson, I. Olsson, M. Arnhold, Y. B i r k , and H. F r i t z (1977). Hoppe-Seyler 1s Z. P h y s i o l . Chem., 358, 53-58.

S c h i e s s l e r , H., 0. Kaplan, S. Wartenberg, and J . Witte (1978). I n t e r ­n a t i o n a l Congress of Inflammation, B o l o g n a / I t a l y , Oct. 31-Nov. 3, a b s t r a c t book p 195.

Schmidt, W. , R. Egbring, and K. Havemann (1974). Thromb. Res. , 6>,315-326.

Witte, J . (1979): Endotoxinämie und hyperdynamer s e p t i s c h e r Schock: Pathobiochemie ausgewählter Gerinnungs- und anderer Plasma-Protein­parameter. H a b i l i t a t i o n S c r i p t , M e d i c a l F a c u l t y of the U n i v e r s i t y of Munich, Munich.

SUBJECT INDEX

Acceptor 529 A c t i v a t i o n of human f a c t o r XII

by plasma k a l l i k r e i n 193 A c t i v a t i o n of p r e k a l l i k r e i n and

f a c t o r XI by f a c t o r XII 193 A c t i v a t i o n products 179 Ac t i v e s i t e m o d i f i c a t i o n 153 Abnormal fibrinogens 51 A c t i v i t y losses 409 Acute lymphoblastic leukemia 523 A f f i n i t y chromatography 299 Agrobacterium tumefaciens 69 Alcohol dehydrogenase muta-

tions 157 A l l i g a t o r ovoraucoid 123 Alloantigens 183 Alpha-antiplasmin 345, 361

365, 379, 413 A l p h a ^ - a n t i t r y p s i n 129 Alpha-chymotrypsin-

i n h i b i t o r s 357 Alpha c r y s t a l l i n 161 Alpha lactalbumin 149 Alpha -macroglobulin 345 Amidolytic and f i b r i n o l y t i c

determination 405 Amino a c i d composition 37 Amino a c i d sequence 23, 47, 51,

69, 75, 83, 87, 99, 193 6-Aminohexandic acid 365 Antibodies 23, 29 Antibody d i v e r s i t y 569 Antibody r e p e r t o i r e 559, 565 Anti g e n i c d r i f t and s h i f t 143 Ant i g e n i c V a r i a t i o n 579 Antigen presentation 599

Antigen p u r i f i c a t i o n 463 Antigen q u a n t i t a t i o n 463 Antigens 537 Antigen s p e c i f i c helper T c e l l

f a c t o r 545 Antigen s p e c i f i c suppressor

fac t o r s 451 Antigen s p e c i f i c suppressor

c e l l hybrids 451 A n t i i d i o t y p e 483, 603 Antibody a f f i n i t y 471 Antibody-binding assay 475 Antibody competition t e s t s 471 Anticoagulant 341 Anticoagulation 249 Antihuman fi b r i n o g e n a n t i -

serum 329 Anti-PA monoclonal antibo­

dies 587 Antiplasmin 395 Antithrombin - I I I 129, 345,

349, 353 Antithrombin - I I I i n Japanese

family 349 Apoprotein I I I 241 Apr o t i n i n 383, 395 Aut o c o r r e l a t i o n 307 Autoimmune disease 583 Avian ovomucoids 123 Azurin 87

B a c t e r i a l cytochromes C 61 Bence-Jones p r o t e i n 3 Beta galactosidase 489 Beta-2-microglobulin 183

611

612 SUBJECT INDEX

Beta propiolactone 229, 333 B i o l o g i c a l antithrombin I I I

a c t i v i t y 349 Biosynthesis 241 Blood coagulation 265, 277 Blood f i b r i n o l y t i c a c t i v i t y 387 Bone marrow t r a n s p l a n t a t i o n 523 Bradykinin 213

Caiman 79 Calcium binding 257, 311 Calcium cycle 95 Calcium ions 321 Cancer markers 587 Carbohydrate 299 Gamma carboxy-glutamic a c i d

249, 253 Carboxylase 249 Carcinoembryonic antigen 501,

511 Casein 421 CEA 517 C e l l attachment 479 C e l l p r o l i f e r a t i o n 205 C e l l - s u r f a c e antigens 475 C e l l u l a r radioimmunoassays 467 Ceruloplasmin 83, 87 Chicken major h i s t o c o m p a t i b i l i t y

complex 183 Chimpanzee 229 C l i n i c a l 541 Cloned 447 CNBr-fragment pattern 103 C0 2 hydratase 153 Coagulation f a c t o r s 333 C o l l o i d a l gold 303 Common molecular ancestor 49 Complement components 179 Computer methods 17 Conditioned medium from thymo-

cytes 459 Conformational determinants 489 Congenital d e f i c i e n c y 349 Contact a c t i v a t i o n of coagula­

t i o n 213, 217, 375 Convergent and divergent evo-

l u t i o n 123 Cross r e a c t i n g idiotypes 493, 565 Cross R e a c t i v i t y 559 C r a y f i s h t r y p s i n 115 Cytochrome C 57, 575 Cytochrome C oxidase 87 Cytochrome C sequence classes 63 Cytochrome C-552 69 Cytochrome C-556 69 Cytochromes C 2 and C 1 69

C y t o l y t i c 447 Cytotoxic T lymphocytes 545 C l 1 - e s t e r a s e i n h i b i t o r 345

D e d i f f e r e n t i a t i o n 3 D e f i b r i n a t e d plasma 329 Detecting r e l a t i o n s h i p s 17 Dextran s u l f a t e 225 D i f f e r e n t i a l l a b e l l i n g 149 Disseminated Intravascular Coa­

g u l a t i o n (DIC) 417 D i s u l f i d e bonds 51 Disulphide bridge 497 Disulphide Oxydation 421 Disulphide reduction 421 DNA sequence 23 Domains 29 Double producer 607 D - v a l y l - L - l e u c y l - L - l y s i n e - 4 -

n i t r o a n i l i d e 383

Electrochemistry 277 E l e c t r o p h o r e t i c v a r i a n t s 157 E l e c t r o n microscopy 307 E l i s a 483 E l l a g i c a c i d 209 Epitope s p e c i f i c i t y 471, 501 E q u i l i b r i u m constant 463 Enzymo-immuno-assay 603 3 epoxide 253 E v o l u t i o n 169 E v o l u t i o n of a c t i v e s i t e s 153 E v o l u t i o n of prokaryotes 61 Evolutionary rate 133 Evolutionary trees 17 Evolutionary v a r i a b i l i t y 47 Eye lens 161

Factor V 341 Factor V d e f i c i e n t plasma 285 Factor VII 265 Factor V I I a A 237 Factor VII a c t i v a t i o n 245 Factor VIII 289, 341 Factor VIII

- chemical composition 299 - Polyacrylamide gel e l e c -

trophoresis 295 - s i z e 303

Factor VHI-Von Willebrand f a c t o r 295, 299

Factor IX 229, 233 Factor IXa 245, 265 Factor X 233

SUBJECT INDEX 613

F a c t o r Xa 245, 273 F a c t o r X a c t i v a t i o n 265 F a c t o r X I 197, 209, 217, 221 F a c t o r X I I 197, 209, 217, 221,

225, 337, 375 F a c t o r X I I

- amino a c i d sequence 193 - i s o l a t i o n 193

F a m i l i a l d e f i c i e n c y 349 FDP 329 F e e d e r c e l l s 443 F e r r i t i n s 91 F e r r i t i n - human and h o r s e 91 F e r t i l i z a t i o n 357 F i b r i n 321, 325, 383, 421 F i b r i n - b i n d i n g 365, 383 F i b r i n o f m a g n e t i c o r i e n t a t i o n 317 F i b r i n p o l y m e r i z a t i o n s i t e s 311 F i b r i n s t r u c t u r e 317 F i b r i n o g e n 47, 51, 307, 317, 321,

325, 333 F i b r i n o g e n p r o t e i n s t r u c t u r e 307 F i b r i n o g e n and f i b r i n o l y s i s

e v o l u t i o n 41 F i b r i n o l y s i s 337, 361, 375

391, 395, 427 F i b r i n o l y s i s i n h i b i t o r s 337,

427 F i s h eggs 357 F r a c t i o n a t i o n by g e l f i l t r a -

t i o n 173 Fragment 1 257, 261 Fragment 2 261 Fragment D 321 F r i g e n 329 F u n c t i o n a l T c e l l h y b r i d s 451 F u s i o n e f f i c i e n c y 443

G a l a c t o s y l t r a n s f e r a s e 149 Gene d u p l i c a t i o n 17, 29, 41,

139, 153, 169 Gene e v o l u t i o n 83 Gene i n t e r a c t i o n 33 Genes 23 G e n e t i c s 33 G e r m l i n e a n t i b o d y s e t 559 G l o b i n s 57 G l u c o s e u p t a k e and m e t a b o l i s m

205 G l u p l a s m i n 383 G l u t a r a l d e h y d e - a c t i v a t e d p l a s t i c

cups 479 G l y c o p r o t e i n 133 G l y c o s y l a t i o n s i t e s 51 G r a n u l o c y t i c p r o t e i n a s e s 417 Growth r a t e / f r a c t i o n 3

H-2-complex 529 H - 2 - d e t e r m i n a n t s 579 H-2-polymorphism 579 H a e m o p o i e t i c p r e c u r s o r (stem)

c e l l s 523 Hageman f a c t o r 201, 337 H a p t o g l o b u l i n 99 H e l p e r c e l l 541 H e m a g g l u t i n i n 143 H e m o g l o b i n 79 H e p a t i t i s B v i r u s 229 H e t e r o c l i t i c r e a c t i o n s 559 Hexose c o n t e n t 37 H i g h mol wt k i n i n o g e n 197, 209

217, 225 H i s t i d i n e r i e h p e p t i d e 213 H i s t o c o m p a t i b i l i t y a n t i g e n s

17 HLA a n t i g e n s 463 HLA-DR a n t i g e n s 587 Homology 99 Human c h o r i o n i c g o n a d o t r o p i n

499 Human chromosome 6 587 Human e n d o t h e l i a l c u l t u r e

s u p e r n a t a n t (HECS) 443 Human g r o w t h hormone 471 Human H i g h M o l e c u l a r Weight

(HMW) k i n i n o g e n 213 Human I a - l i k e i s o t y p e s 595 Human melanoma a n t i g e n s 505 Human p l a s m a 427 H y b r i d c e l l s 517 Hybridomas 443, 489, 537, 583,

587, 595, 603 H y b r i d o m a - d e r i v e d - a n t i - a z o b e n -

z e n e - a r s o n a t e - a n t i b o d i e s 493 Hybridoma S c r e e n i n g t e s t s 533 H y p e r a g g r e g a b i l i t y o f p l a t e ­

l e t 349 H y p e r v a r i a b l e r e a c t i v e s i t e s

123 H y s t r i c o m o r p h s 133

I a - a n t i g e n s 599 I d i o t y p e s 569, 603 I d i o t y p e - a n t i i d i o t y p e i n t e r a c -

t i o n s 575 I d i o t y p e s u p p r e s s i o n 565 I d i o t y p i c n e t w o r k 483, 603 IgM, b e t a 2 m 433 Immune r e g u l a t i o n 451 Immune r e s p o n s e t o p h o s p h o r y l -

c h o l i n e 565 Immunochemical and c h r o m o g e n i c

S u b s t r a t e a s s a y s 345

614 SUBJECT INDEX

Immunoglobulin M mutants 439 Immunoglobulin v a r i a b l e region

determinants 545 Immunoglobulins 17, 23, 607 Immunoglobulins i n semen 173 Immunological antithrombin I I I

concentration 349 Immunoperoxidase procedures 475 Immunotoxins 497 Influenza v i r u s 559 Influenza A v i r u s 143 I n h i b i t i o n assays 467 I n h i b i t o r f a m i l i e s 123 I n h i b i t o r protease complexes 379 I n h i b i t o r s p e c i f i c i t y 107 I n s u l i n - l i k e growth f a c t o r 165 I n t e r r e l a t i o n s h i p of t i s s u e hormo-

nes 205 Internal homology 83 I n t r a c e l l u l a r calcium binding

proteins 95 Introns 123 Invertebrate proteases 115 Invertebrate proteinases 107 In v i t r o immunization and

h y b r i d i z a t i o n 459 In v i t r o f u n c t i o n a l Systems 549 Ir-genes 599 I s o e l e c t r i c focusing 353 Isozyme evolution 153

J-segment 33

K a l l i k r e i n 201, 221 K a l l i k r e i n - k i n i n o g e n - k i n i n -

kinase Systems 205 K chains 33 K i n e t i c s 325 K i n i n 201 Kringle structures 371

Laccase 87 Lactose synthase 149 Laser nephelometer 329 Levan 483, 603 Light s c a t t e r i n g 325 Low molecular weight pro­

tease 107 Low molecular weight dextran

sulphate 413 Lys-plasmin 383 Lysine binding s i t e s 371

Mammals 133, 139 Man-mouse hybridomas 433 Maximum parsimony reconstruc-

t i o n 95 Mg and f i b r i n o l y s i s 427 Miniplasminogen 365 Molecular evolution 61, 69 Molecular v a r i a n t s 51 Monoclonal antibodies 463, 471,

475, 497, 501, 505-, 511, 517, 523, 537, 541, 549, 553, 559, 565, 575, 599, 607

Monoclonal a n t i - i d i o t y p e s 569 Monocytes 241 M u l t i p l e tandem domains 123 Murine 447 Muscle aldolase 111 Muscle carbonic anhydrase 153 Mutation frequency 439 Myeloma 3 Myosin 75 Myosin l i g h t chain L27 75

N-carbobenzoxy-tyrosine-ni t r o p h e n y l - e s t e r 421

NAD binding domain 157 Natural s e l e c t i o n 57 Nerve Growth f a c t o r 165 Non-proteolytic bapna-ase 357 Nu/Nu mice 533 Nuclear Magnetic Resonance

(NMR) 261 Nucleic a c i d sequences 17 Nucleotide sequence 583 NZB/NWZ mice 583

Oriented f i b r i n gels 317 Ovoinhibitor 123 Oxygen a f f i n i t y 79

Pancreatic secretory t r y p s i n i n h i b i t o r 123

Paraprotein 3 Paratopic s p e c i f i c i t y 559 Phenotype a s s o c i a t i o n 579 Phospholipase A2 281 Phospholipid monolayers 277 Phospholipids 265, 281 Phosphorylcholine antibodies 569 Phylogeny 161 P i g heart 75 Plasmin 221, 379, 395, 421

SUBJECT INDEX 615

Plasmin - a f f i n i t y chromatography 399 - a p r o t i n i n - b i n d i n g

domain 399 - a p r o t i n i n complex 383 - conformation 399 - derived Light chain 399 - d i g e s t i o n 321 - i n h i b i t o r s 357

Plasminogen 361, 365, 371, 383 Plasminogen

- a c t i v a t o r s 361, 375, 427, 487 - bovine and porcine 103 - chemical modifications 371 - fragments - elastase 365 - human, bovine and porcine 103

Plastocyanin 87 P l a t e l e t s 273 Polymorphism 139, 353 P r e k a l l i k r e i n 197, 217, 221,

225 Primary structure 79, 91, 103

111, 213 P r i v a t e H2 antigens 579 Prorenin 221 P r o s t a t i c a c i d Phosphatase 501 Protease i n h i b i t o r s 379 P r o t e i n a spot test 533 P r o t e i n C 341 P r o t e i n evolution 23, 87, 111,

161 P r o t e i n i n t e r a c t i o n s 149 P r o t e i n oligomerization 169 P r o t e i n - p r o t e i n i n t e r a c t i o n s 489 P r o t e i n sequences 17, 575 Proteinase i n h i b i t o r 417 Proteinases 357 P r o t e o l y t i c degradation 295 P r o t e o l y t i c s p e c i f i c i t y 107 Prothrombin 253, 257, 273, 277,

285 Prothrombin

- a c t i v a t i o n 265, 281 - bovine 261 - i n h i b i t i o n 281 - synthesis 249

P u b l i c H-2 antigens 579

Quaternary structure 29

Rabbit 33 R a d i o a c t i v i t y 277 Radioimmunoassay 91, 511, 517 Rates of molecular evo l u t i o n 57 Reaction k i n e t i c s 467

Receptor 169 Receptor Substrate i n t e r a c t i o n

169 Recombinant DNA technology 143 Relaxin 165 Release 273 Renin 221 R e p t i l i a n IgM 37 Review sequence data 61 Ribonuclease 133, 139 Ricin-A-chain 497 RNA 583

Salmonidae 357 Second messenger calcium 95 Secondary hormones 165 Secondary structure p r e d i c t i o n

157 Seminal plasma proteins 173 Septicemia 417 Sequence comparisons 33, 41,

103 Sequence homology 47, 119, 165

213 Serine proteases 99, 115, 119,

295 Serine Proteinase i n h i b i t o r s

129 S i l e n t r e p e r t o i r e 565 Skink C3 179 SLE 583 Solid-phase immunoassay 479 Sonratic antibody set 559 Somatic recombination 439 S p e c i f i c target c e l l k i l l i n g 545 Spleen lymphocytes 183 S t e r i l i z a t i o n 229 Streptococcal g r o u p - s p e c i f i c

antibodies 459 Streptokinase 395 4-substituted glucose 149 Substrate i n h i b i t o r 421 Sulfonamide i n h i b i t i o n 153 Suppressor c e l l 541 Suppressor f a c t o r 529 Synthetic peptide a i i t i p o l y -

merants 311

T - c e l l 537 T - c e l l d i f f e r e n t i a t i o n 553 T - c e l l hybrid 529 T-cell-hybridomas 447, 545 T - c e l l - s u b s e t s 549 Theory of blood coagulation 285 T h i o l groups 111

616 SUBJECT INDEX

Thrombin 289, 325 Thrombin i n h i b i t o r y a c t i v i t y 349 Thrombolysis 361 Throrabotic disease 337 Thymocytes 523 T i l i q u a rugosa 37, 179 Tissue f a c t o r 233, 237 Tissue Plasminogen a c t i v a t o r

Tissue s p e c i f i c i t y 91 Tissue thromboplastin 241 Tranexanic aci d 365 Transport proteins 169 Trypsin evolution 115 T r y p s i n - i n h i b i t o r s 357 T r y p t i c peptides 111 Tumor membrane antigens 533 Tumor marker 517 Two dimensial electrophoresis

Urokinase 221, 387, 391, 405 409, 413

Urokinase - a c t i v i t y 405 - molecular d i s t r i b u t i o n 409 - p u r i f i c a t i o n 409

V a r i a b i l i t y of antibody production 439

Vascular Plasminogen a c t i v a t o r 387, 391

Vitamin K 249, 253 Vitamin K 2 253 Vitamin K-dependent carboxylase

Vitamin K epoxidase 253

Warfarin 249, 349

X-Ray crystallography 257

Zymogens 237

387

285

253

r Bayerische Staatsbibliothek

München