PROTIDES OF THE BIOLOGICAL FLUIDS

PROCEEDINGS OF T H E TWENTY-NINTH COLLOQUIUM, 1981

Edited by

H.PEETERS

D i r e c t o r I n s t i t u t e for M e d i c a l Biology

Brüssels, B e l g i u m

PERGAMON PRESS

O X F O R D • N E W Y O R K • T O R O N T O • P A R I S • F R A N K F U R T • S Y D N E Y

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f Bayerische I Staatsbibliothek l München J

P r i n t e d i n G r e a t B r i t a i n by A . Wheaton & C o . Ltd., E x e t er

ISBN 0 08 0279880 ISSN 0079-7065

CONTENTS

P R E F A C E xxv

A C K N O W L E D G E M E N T S xxvii

T H E T E N T H ARNE TISELIUS MEMORIAL LECTURE

H - Y Antigen and Sex Determination 3 H A R T W I G C L E V E

SECTION A. MEMBRANE PROTEINS

A . l . Pure Membrane Proteins

A . l . l . Purification Procedures

Some General Aspects of the Difficulties to Purify Membrane Proteins 15 S. H J E R T E N , H . P A N and K . Y A O

New Application of Group-Specific Reagents for Hydrophobie Labeling and Crosslinking of Membrane Proteins 27 H . SIGRIST, P . R. A L L E G R I N I , A . H A L D E M A N N and P . Z A H L E R

The Use of a Zwitterionic Detergent and a Monoclonal Antibody in Purification of Rat Liver 5'-Nucleotidase 33 J . P . L U Z I O , E . M . B A I L Y E S , A . C . N E W B Y and K . S I D D L E

v

vi C O N T E N T S

A . 1.2. Immunoglobulins

Human IgD, The Gene and the Protein 37 T. H . R A B B I T T S and C . P . M I L S T E I N

Structural Studies of Human IgD 41 B . D E B U I R E and F . W . P U T M A N

Comparison of Secreted and Membrane-Bound Human Immunoglobulins M and D 45 R. K . L . B I J L E N G A , H . P . K O C H E R and J . -C . J A T O N

A.1.3. Glycoproteins

The Sialoglycoproteins (Glycophorins) in the Plasma Membranes of Different Cells and Their Receptor Functions 49 H . F U R T H M A Y R

Structures and Antigenic Properties of Human Erythrocyte Membrane Sialoglycoproteins 57 W . D A H R , K . B E Y R E U T H E R , E . B A U S E and M . K O R D O W I C Z

Cytoskeleton-Associated Surface Glycoproteins of Murine Lymphocytes 63 E . RUNGGER-BRÄNDLE and D . C . H O E S S L I

The Relationship between Differentiation of Friend Erythroleukemia Cells and Synthesis of Plasma Membrane-Bound Glycoproteins 67 G . J . C . M . B O S M A N , P. B O E R and E . P . S T E Y N - P A R V E

Variations in the Structure of Surface Glycoproteins Present in Different Lymphocyte Subpopulations 71 D . H O E S S L I , J . R. L . P I N K , R. H O O G H E , M . S C H R E I E R and P . V A S S A L L I

Enhanced Neuraminidase-Resistant Sialic Ac id Content in the Surface Glycopeptides of Human Leukaemic Hairy Cells 75 G . A . J . M . V A N D E R H O F S T A D and M . E . V A N R Y M E N A N T

Reconstitution of Liposomes Bearing Platelet Membrane Glycoproteins. A Functional Study with Von Willebrand Factor 79 P . S l f i , M . G I L L O I S , H . J . C H A P , B . B O N E U , R. B I E R M E and L . D O U S T E - B L A Z Y

Human Platelet Membrane Glycoproteins IIb and l i l a May form C a 2 + -Dependent Complexes 85 D . P I D A R D , T. K U N I C K I , J . -P. R O S A and A . T. N U R D E N

Glycoproteins of the Human Syncytiotrophoblastic Plasma Membrane M . J . O ' S U L L I V A N , J . A . M c I N T Y R E , G . A . W A R R I N E R and 89 W . P A G E F A U L K

C O N T E N T S vii

Characterization of a-Tissue Antigen (aTA), A Membrane Glycoprotein of Cancer Cells 93 A . Y A C H I , M . K A W A H A R A D A , Y . A K A H O N A I , K . I M A I and T. W A D A

The Structure and Function of a Membrane Glycoprotein Encoded in the Adenovirus Genome 97 H A K A N P E R S S O N and L E N N A R T P H I L I P S O N

Protein Micelles and Virosomes from the Surface Glycoproteins of Parainfluenza 3 Virus 101 B. M O R E I N , B . S U N D Q U I S T , S. H Ö G L U N D , A . H E L E N I U S and K . S IMONS

A.1.4. Erythrocyte Membrane Proteins

Human Red Cell Membrane Acetylcholinesterase, A Model Enzyme for the Study of Protein-Amphiphile Interactions 105 U . B R O D B E C K and P. O T T

Spectrin Free Vesicles Obtained from Human Erythrocytes Without ATP-Deplet ion 109 P . O T T and U . B R O D B E C K

Acylation of Integral Erythrocyte Membrane Proteins Resulting in a Soluble Form of Band 3 Protein 113 F R A N Z H E R B S T and V I C T O R R U D L O F F

Self-Association of Band 3 Protein from Erythrocyte Membranes in Solutions of a Nonionic Detergent, Ammonyx-LO 117 G U N T E R P A P P E R T and D I E T E R S C H U B E R T

Interactions of the Isolated Spectrin Polypeptides (Bands 1 and 2) with Phospholipid Monolayers 121 D I E T E R S C H U B E R T , F R A N Z H E R B S T , H A N N E L O R E M A R I E and V I C T O R R U D L O F F

Red Cell Membrane Changes in a Heinz Body Anaemia (Kaie Anaemia) 125 R. H . S M I T H , W. B . W A T T , W . J . L A W S O N and C . R I C E - E V A N S

Ozone-Induced Cross-Linking of Erythrocyte Membrane Proteins 129 H . V E R W E I J and J . V A N S T E V E N I N C K

A.1.5. Various Cell Membrane Proteins

Membrane Proteins of Murine T Cell Clones C . B R O N , M . S C H M I D , G . C O R R A D I N , A . G L A S E B R O O K , H . E N G E R S , 133 C. H O R W A T H , J . C . C E R O T T I N I and H . M A C D O N A L D

viii C O N T E N T S

Isolation and Characterization of Basal-lateral and Luminal Plasma-Membranes (PM) from the Proximal Tubule of Human Kidney 139 J . E . S C H E R B E R I C H , R. K I N N E , C . G A U H L , W. M O N D O R F and W . S C H O E P P E

Further Immunochemical Studies of Tissue Polypeptide Antigen (TPA) 143 BO W I K L U N D and BJÖRN LÜNING

Purification of Membrane 5'Nucleotidase Preparation and Properties of Anti-5 ' Nucleotidase Serum 147 J . D O R N A N D , J . C . B O N N A F O U S and J . C . M A N I

Characterization of Four Chick Brain Synaptic Membrane Proteins 151 M . J . A L B E C K and E . B O C K

Isolation of a Membrane Antigen from Electric Organ of T o r p e d o C a l i f o r n i c a 155 E . B O C K and A . C . v. G E R S T E N B E R G

Characterization of Some Antigens Associated with the Membrane of Hampster Tumor Cells 159 A . D U T H U , I. A L E X A N D R O V , H . H A D D A D A , N . L E G O F F I C , J . Z U I N G H E D A U and C H . D E V A U X S A I N T C Y R

Non-Specific Binding of (-) 3H-Dihydroalprenolol to Intact Human Lymphocytes and Lymphocyte Membranes 163 H E R M A N M E U R S , W I L L E M V A N D E N B O G A A R D , H E N K F. K A U F F M A N and P I E T E R L . B . B R U Y N Z E E L

Characterization of (-) ( 3 H) Dihydroalprenolol Binding Sites on Intact Human Peripheral Blood Lymphocytes 167 P . L . B. B R U Y N Z E E L , W . V A N D E N B O G A A R D , M . L . H A M E L I N K and J . A . M . R A A I J M A K E R S

A.1.6. Bacterial Membrane Proteins

Structure and Function of a Pore-Forming Transmembrane Protein: High Resolution Studies of a Bacterial Porin 171 J . P . R O S E N B U S C H , R. M . G A R A V I T O , D . L . D O R S E T and A . E N G E L

Evidence for Protein-Lipid Interactions in Membranes of A c h o l e p l a s m a L a i d l a w i i A . W I E S L A N D E R , K . - E . J O H A N S S O N , C . JÄGERSTEN and 175 A . C H R I S T I A N S S O N

Isolation of Surface Antigens from Schistosoma Mansoni Schistosomula 179 C. DISSOUS and A . C A P R O N

Crossed Immunoelectrophoretic Analysis of the Purple Membrane of H a l o b a c t e r i u m H a l o b i u m H . WRÖBLEWSKI and G . B A F F E T

183

C O N T E N T S ix

A.2. Cell Surface Antigens

A.2.1. Beta 2 Microglobulin

Beta-2-Microglobulin and its Association with Histocompatibility Antigens of the Mouse and Rabbit 187 T. J . K I N D T , J . E . C O L I G A N , F . T. G A T E S III, E . S. K I M B A L L , W . L . M A L O Y and J . M . W I L K I N S O N

Evolutionary Conservation of the Interaction between ß 2 -Microglobulin and ß 2 -Microglobulin-Binding Molecules 191 L . LÖGDBERG, L . BJÖRCK and R. C I G E N

Seminal Plasma Levels of Beta 2-Microglobulin and C E A - L i k e Protein in Infertility 195 J . L I Z A N A , P. E N E R O T H , B. BYSTRÖM and M . B Y G D E M A N

Stimulation of Human Monocyte Chemotaxis by Anti-ß 2 -Microglobulin 201 T. P. C O N W A Y and M . D . P O U L I K

Exchange of ß 2 -Microglobulin in Histocompatibility Antigens by its Human Analogue 205 W. S C H M I D T , A . R. S A N D E R S O N and H . F E S T E N S T E I N

Rat Serum Components with Affinity for Human ß2 Microglobulin 209 C. V I N C E N T and J . P . R E V I L L A R D

A .2.2. Blood Group Antigens

Human Blood Group M - , N - , T (Thomsen-Friedenreich)-, and Tn-Specific Lipidic Substances in Line-10 Hepatocarcinoma of Strain-2 Guinea Pigs 213 G . F . S P R I N G E R , J . L . C A N T R E L L , P . R. D E S A I and H . T E G T M E Y E R

Further Evidence that Terminal ß-Galactopyranosyl Groups are Immunodeterminant in Blood Group N Specificity 217 G . F . S P R I N G E R and H . T E G T M E Y E R

A .2.3. Histocompatibility Antigens

Biosynthesis of H L A - A and -B Antigens 221 M . J . O W E N , A . - M . K I S S O N E R G H I S and C . K I N N O N

Biochemical Characterisation of Leukaemia-Associated Cell Surface Antigens 227 R. A . N E W M A N , D . R. S U T H E R L A N D and M . F . G R E A V E S

Influence of Selective Absence of H - 2 K K on Gross Virus Leukaemias I n V i t r o and the Implications for I n V i v o Surveillance of Leukaemia 231 H . F E S T E N S T E I N , W . S C H M I D T , A . A L O N Z O and L . L E B E N

X C O N T E N T S

Primary Structural Analyses of Murine Major Histocompatibility Antigens J . E . C O L I G A N , E . S. K I M B A L L , T. J . K I N D T , W. L . M A L O Y , 237 J . M . M A R T I N K O , R. N A I R N , S. G . N A T H E N S O N , H . U E H A R A

Characterization of D L A SD Antigens using Immunochemical Techniques M . J . M . V A N D E R F E L T Z , J. A . G . M . V A N D E R K O R P U T , 243 M . J . G I P H A R T and D. L . W E S T B R O E K

Neuron-Specific Markers: A Critical Appraisal E . L . H O O G H E - P E T E R S , A . W. S I A D A K , C. B E R E K and R. J . H O O G H E 247

Variations in the Antigenic Architecture o f the A c h o l e p l a s m a L a i d l a w i i Cell Membrane 251 K A R L - E R I K J O H A N S S O N and C H R I S T I N A JÄGERSTEN

Interspecies Crossreactivities of Antisera Directed Against (Na,K)-ATPase Subunits from B u f o M a r i n u s 255 M . G I R A R D E T , D . GESER, K. G E E R I N G and C . B R O N

A.3. Membrane Function

A.3.1. Transport Function

Transport of Proteins into Mitochondria H O W A R D R I E Z M A N , SUSAN GASSER, P E T E R BÖHNI, 259 G Ü N T H E R D A U M , A K I R A O H A S H I , J A N E G I B S O N and G O T T F R I E D S C H A T Z

The Stereospecific D-Glucose Transport Activity of Cholate Extracts from Human Erythrocyte Membranes. Effects of Cholate, E D T A and Dithioerythritol 263 P . L U N D A H L , S. P H U T R A K U L , F. A C E V E D O and G . FRÖMAN

Side-Specific Transfer of Membrane-Bound Phospholipase A 2 to Other Membranes by Fusion 267 P . Z A H L E R and M . W O L F

Bile Ac id Transport in Distal Ileum F . W . M . D E R O O I J , J . W. O. V A N D E N B E R G , 271 E . P . B O S M A N - J A C O B S and A . C. B L O M M E S T E I J N

The Kinetics of Intramolecular Crosslinking of the Band 3 Protein by 4,4' -Diisothiocyanato Dihydrostilbene-2,2'-Disulfonic Acid (H 2 DIDS) 275 L . K A M P M A N N , S. L E P K E and H . PASSOW

The Effect of Arginine Specific Reagents on Anion Transport Across Red Blood Cells 279 L A I L A Z A K I

C O N T E N T S xi

Does Ternary Complex Formation Control the State of the Cation Barrier in the Resealed Human Red Blood Cell Ghost? 283 P H I L L I P G . W O O D

A.3.2. Cell Adhesion

Microfilaments and Fibronectin Fibers Involved in the Formation of Fibroblast^ Focal Contacts 287

W . B I R C H M E I E R

Cell Surface Glycoproteins and Fibronectin Mediated Cell Adhesion 291 G . T A R O N E , G . G A L E T T O and P . M . C O M O G L I O

The Actin Network in Human Granulocytes at Different Stages of Phagocytosis in Granulopathies 295 Y . F R O B E R T

Expression of Fibronectin and Plasminogen Activator in Human Cell Hybrids L . L A R I Z Z A , M . T E N C H I N I , A . M O T T U R A , E . R A M P O L D I , M . C O L O M B I 299 and S. B A R L A T I

Role of Fibronectin Degradation Products on Cellular Transformation 303 S. B A R L A T I , G . D E P E T R O , T. V A R T I O and A . V A H E R I

A.3.3 Cell Growth

Possible Role of the Plasma Membrane in Regulating Cell Growth Behaviour 307 K . B A L L M E R and M . M . B U R G E R

Infiltration of Lymphosarcoma Cells into hepatocyte Cultures: Inhibition by Univalent Antibodies Against Liver Plasma Membranes and Lymphosarcoma 313 Cells O. P. M I D D E L K O O P , E . ROOS and I. V . V A N D E P A V E R T

The Role of Components of the Outer Membrane of Gram-Negative Bacteria in the Serum-Bactericidal Effect 317 F. C L A S and M . L O O S

Receptor-Ligand Interaction Leading to Entry of Myxoviruses into Host Cells 321 R I C H A R D T. C . H U A N G and R U D O L F R O T T

A.3.4. Cell Ageing

Molecular Aging: A Termination Antigen Appears on Senescent Cells 325 M . M . B. K A Y

K + Fluxes Düring T Lymphocyte Activation in Human Aging 329 B . K E N N E S , P . M O E S T , F . R I C K A R T , A . D E M U L D E R and P . N E V E

xii C O N T E N T S

Age-Related Defect of Cytokine-Ca 2 + - c G M P Mediated Transmembrane Signal in T Lymphocyte Activation B. K E N N E S , I. D U M O N T , P . P IRO and P. N E V E

Supramolecular Assembly of Membrane Constituents Düring ' I n V i t r o * Aging of Human Erythrocytes A . B R O V E L L I , G . P A L L A V I C I N I , G . A I R O L D I and C . B A L D U I N I

Masking of R C A I Binding Sites by Hyaluronic Ac id in Foetal Liver Cells B. B E R N A R D , J . F O N T , M . A U B E R Y , C . G U G U E N - G U I L L O U Z O and A . G U I L L O U Z O

Parallel Increase of 3-0-Methylglucose Uptake and Cytochalasin B Binding in Chick Fibroblasts Düring Embryo Development B. B E R N A R D and M . A U B E R Y

SECTION B. RECEPTOR LIGAND INTERACTIONS

B . l . Structure Activity Relations

The Acetylcholine Receptor: A n Allosteric Membrane Protein R. E . O S W A L D , T. H E I D M A N N and J . P . C H A N G E U X

Structure and Function of the Human Platelet Von Willebrand Factor Receptor K . J . C L E M E T S O N , H . Y . N A I M and E . F. LÜSCHER

Studies on the Receptor Sites of the Monoclonal Anti-Pr and -Sa Cold Agglutinins W . D A H R , D . L I C H T H A R D T and D . R O E L C K E

Relations Between Serum a r Foetoprotein (AFP) and uterine Cytosol Receptors in the Immature Rat: Importance of Fatty Acids Ligands C. B E N A S S A Y A G , G . V A L L E T T E , L . S A V U , F . C L E R C - H O F M A N N , N . C H R I S T E F F , F . M U L L E R , J . D E L O R M E and E . A . N U N E Z

Specific Binding of the Apo and Holoforms of the Serum D B P to the 'soluble' part of Actin from Human Lymphocytes and Platelets J . C O N S T A N S , M . V I A U and C . G O U A I L L A R D

A C T H Effects on Steroidogenesis and Adrenal Protein Synthesis: Relation Structure-Function A . D A Z O R D and J . M . S A E Z

Similarities of the Gaunine Nucleotide Binding Sites Responsible for Adenylate Cyclase Activation and Carbamylcholine Binding Inhibition, In Rat Heart Membranes M A G A L I W A E L B R O E C K , P I E R R E C H A T E L A I N , P A T R I C K R O B B E R E C H T and J E A N C H R I S T O P H E

C O N T E N T S xii i

Studies on the Hydrophobie Anchor of the Pig Intestinal Intrinsic Factor Receptor 385 I. K O U V O N E N and R. GRÄSBECK

Receptor-Mediated Endocytosis of Glycosylated Derivatives of Bovine Serum Albumin: Targeting Based on Sugar Recognition 389 J . Q U I N T A R T , J . L I M E T and P . B A U D H U I N

Structure-Activity-Relationships in the Steroidal 17ß - Carboxamide Series 393 P. F O R M S T E C H E R , P . L U S T E N B E R G E R and M . D A U T R E V A U X

Binding of Cathepsin B by Haptoglobin 397 D . B U R N E T T , N . K A L S H E K E R and A . R. B R A D W E L L

B.2. Immunoglobulin Receptors

Regulation of the Immune Response by the Fe Portion of Immunoglobulin E D W A R D L . M O R G A N , M A R I L Y N L . T H O M A N and 401 W I L L I A M O. W E I G L E

Human T-Lymphocytes' Fc-Receptors with High Affinity for Monomeric IgG 405 M . R. H A D A M , J . G . S A A L and H . E . F E U C H T

Isolation of Fcy Receptors Released by Human Lymphocytes 409 C . S A M A R U T , L E T H I B I C H T H U Y and J . P . R E V I L L A R D

Cyanogen Bromide Fragments of Mouse IgG2a and IgG2b which Bind to their Fe Receptors on Mouse Macrophages 415 B. D I A M O N D , R. C A M P B E L L , J . S C H N E C K and B . K . B I R S H T E I N

Anti-Receptor Antibodies: A Comparison in the Rat of the Plasma to Bile Transfer of Purified IgG, F(ab') 2 and Fab' Antibodies Against Rat Secretory 419 Component (SC) I. L E M A I T R E - C O E L H O , R. M E Y K E N S and J . P . V A E R M A N

Receptor-Mediated Endocytosis of Polymerie IgA by Cultured Rat Hepatocytes 423 J . N . L I M E T , Y . - J . S C H N E I D E R , J . P . V A E R M A N and A . T R O U E T

A Neo-Antigen Present on Complex a 2 M , Related to the Receptor Recognition Site 427 P . M A R Y N E N , F . V A N L E U V E N and J . J . C A S S I M A N

Events Initiated by Soluble Immune Complex Interaction with Immunoglobulin Receptors on Macrophages 431 R. G . Q. L E S L I E

xiv C O N T E N T S

B.3. Complement Receptors

Molecular Weight Analysis of Human Complement (C3) Receptors Isolated by Immunoprecipitation 435 J . G E R D E S and H . S T E I N

The Receptor Functions of Endogeneous C l q , A Subcomponent of the First Component of Complement, on Peritoneal Macrophages 439 M . L O O S and H . U . S C H O R L E M M E R

Receptors for C3b, C3bi, C3d and ß l H on Lymphocytes and Phagocytic Cells M . P . D I E R I C H , K . P. H A M M A N N , H . - H . M U S S E L , M . S C H M I T T and 443 O. S C H E I N E R

B.4. Transferrin and Herne Receptors

Evidence for Endocytosis of Iron Loaded Transferrin by Cultured Rat Embryo Fibroblasts 447 J . N . O C T A V E , Y . - J . S C H N E I D E R , R. R. C R I C H T O N and A . T R O U E T

Transferrin Receptors in Transformation and Differentiation 451 C H A N G - J I N G G . Y E H and W . P A G E F A U L K

Mediation of a Receptor Mechanism in the Uptake of Iron from Transferrin by the Hepatocyte 455 R. M . N U N E S , J . M . P R I E T O and B . J . P O T T E R

Iron Uptake by Chinese Hamster Fibroblasts from Human Transferrin 459 S. SMIT, M . I. E . H U I J S K E S - H E I N S and B . L E I J N S E

Receptor-Mediated Endocytosis of Heme-Hemopexin by Rat Hepatocytes Y . - J . S C H N E I D E R , J . N . L I M E T , C . S L A C H M U Y L D E R - O T T E and 463 A . T R O U E T

Spectral and Other Studies on the Intestinal Herne Receptor of the Pig 467 R. GRÄSBECK, I. K O U V O N E N and R. M A J U R I

B.5. Neurochemical Receptors

Differentiation between Stereospecific Binding Sites on a Dopamine Receptor Complex: Effects of Proteolytic Enzymes 471 J . E . L E Y S E N and P . V A N G O M P E L

Biochemical and Pharmacological Studies of Substance P Receptors M . H A N L E Y , C . L E E , S. W A T S O N , B. S A N D B E R G and L . I V E R S E N

477

C O N T E N T S xv

Neurotensin-Receptor Interactions: Binding and Pharmacological Studies; Biological Relevance; Structure-Activity Relationships 483 P. K I T A B G I

A Novel Triazolopyridazine, 3 H - C L 218, 872 which Labels the Benzodiazepine Receptor in Rat Brain 487 H . I. Y A M A M U R A , T. M I M A K I , M . M O R E L L I , S. H . Y A M A M U R A , W . D . H O R S T , G . B A U T Z and R. O ' B R I E N

Immunological Studies of ß Adrenergic Catecholamine Receptors: A Two-Way Approach P . O . C O U R A U D , A . B . S C H R E I B E R , C . D E L A V I E R - K L U T C H K O , 493 C . A N D R E , O. D U R I E U - T R A U T M A N N , B . V R A Y , A . S C H M U T Z and A . D . S T R O S B E R G

Asymmetry of Acetylcholinesterase and Acetylcholine Receptor in Intact Secretory Vesicles from Adrenal Medulla 497 M . G R A T Z L and H . K R I E G E R - B R A U E R

Loss of ß 2 -Adrenoceptors After Haemophilus Influenzae Vaccination 501 G . K . T E R P S T R A , J . A . M . R A A I J M A K E R S and J . K R E U K N I E T

Characterization of Adrenergic Binding in Canine Lung Tissue 505 J . A . M . R A A I J M A K E R S and G . K . T E R P S T R A

B.6. Vasoactive Intestinal Polypeptides (VIP)

Heterogeneity of VIP-Secretin Receptors in Three Rat Tissues P A T R I C K R O B B E R E C H T , P I E R R E C H A T E L A I N , M A G A L I W A E L B R O E C K 509 and J E A N C H R I S T O P H E

Vasoactive Intestinal Peptide (VIP) Control of Cyclic A M P Accumulation in Digestive Epithelia: Involvement of Multiple Regulatory Processes 513 M . L A B U R T H E , B . A M I R A N O F F , C . D U P O N T , C . G E S P A C H and G . R O S S E L I N

Effects of G T P on Vasopressin, and Angiotensin Binding to Rat Liver Membranes 517 S. J A R D , P . C L A U S E R , R. B O N O and B. C A N T A U

Cholecystokinin in Brain and Hypophysis J . J . V A N D E R H A E G H E N , F . L O T S T R A , G . V I E R E N D E E L S , 521 C . D E S C H E P P E R , C . G I L L E S and P . V E R B A N C K

Vasoactive Intestinal Polypeptide (VIP): Specific Receptors on Smooth Muscle Membranes from Porcine Uterus 527 B . O T T E S E N , P . S T A U N - O L S E N , S. G A M M E L T O F T and J . F A H R E N K R U G

xvi C O N T E N T S

Impaired Secretin-VIP Sensitivity of Adenylate Cyclase in the Heart of Congenitally Hypertensive (SHR) or Obese (fa/fa) Rats 531 P I E R R E C H A T E L A I N , P A T R I C K R O B B E R E C H T and J E A N C H R I S T O P H E

B. 7. Hormone Receptors

B.7.1. Insulin Receptors

Multiple Functions of Peptides: Insulin and Vasoactive Intestinal Polypeptide Bind to Receptors in Liver and Brain 535 S. G A M M E L T O F T , P . S T A U N - O L S E N , J . F A H R E N K R U G and B . O T T E S E N

The Complex Kinetics of Insulin Binding to Receptors on Isolated Pig Hepatocytes 539 S. G A M M E L T O F T , P . S T A U N - O L S E N , J . F A H R E N K R U G and B. O T T E S E N

I n V i v o Studies by Gamma-Camera of 1 3 ^-Insulin Distribution and Metabolism in Rabbits 543 P . B O U R G E O I S , F. L E G R O S , M . L A N G O H R , A . V E R B I S T , G . G H A N E M and J . FRÜHLING

Immunohistochemical and Autoradiographic Visualization of Porcine and Human 1 2 5 I-Insulin Hepatic Binding I n V i v o 547 P . S T E E N H O U T , M . G U I L L A U M E , F. B I E R L A I R E , G . G H A N E M and F. L E G R O S

B.7.2. Alpha MSH

Variation of a - M S H Receptivity Between Malignant Melanocytes from Horizontal and Vertical Growth Phases in a Human Superficial Spreading Melanoma 551

F. L E G R O S , J . C O E L , A . G O S S U I N and F . J . L E J E U N E

1 2 5 I - a - M S H Binding in Human Melanoma Cell Lines A . G O S S U I N , G . G H A N E M , A . T E M M E R M A N , J . C O E L , P . E . H E N R Y 555 and F . L E G R O S

D N A Replication and Production of Retroviral Markers in Human Melanoma Cultures Depends on Degree of Differentiation and on Hormonal Stimulation 559 N . V A N T I E G H E M , A . V E R C A M M E N - G R A N D J E A N , M . F O O I J , A . T E M M E R M A N and F . J . L E J E U N E

Binding of Endogenous a - M S H to Malignant Melanocytes in Human Melanomas A . N U C H O W I C Z , A . V E R H E S T , A . G O S S U I N , S. L E G R O S , A . L I T E A N U , 563 P . H A N S O N , A . S O U P A R T , F . L E G R O S and F . J . L E J E U N E

Hormonal Screening in Patients Suffering Malignant Melanoma. A Preliminary Approach 567 M . L A N G O H R , C . R O B Y N , F . L E G R O S and F . J . L E J E U N E

C O N T E N T S xvii

B.7.3. Other Hormones

Antibodies Against T S H Receptor in Graves' Disease: Evaluation by Release of c A M P 571 J . M . GÖMEZ, M . A . N A V A R R O , M . C . P A S T O R and J . S O L E R

Antibodies Against T S H Receptor in Graves' Disease: Relation between c G M P / c A M P Release ; 575 M . A . N A V A R R O , J . M . G Ö M E Z , M . C . P A S T O R and J . S O L E R

Negative Controls of Dog Thyroid Adenylate Cyclase Cyclic A M P System 579 P . C O C H A U X , J . V A N S A N D E and J . E . D U M O N T

Rat Liver Glucocorticoid Receptor Purified in Presence of Molybdate 583 P . L U S T E N B E R G E R , P . F O R M S T E C H E R , M . D A U T R E V A U X

Induction of Prolactin Liver Receptors in the Male Rat P . G O O N E W A R D E N A , P . E N E R O T H , A . M O D E , G . N O R S T E D T 587 and J . Ä. G U S T A F S S O N

A n Affinity Adsorbent for an Estrogen Binding Protein Rat a-Fetoprotein 591 R. Z E E U W S , V . V E R S E E and A . O. B A R E L

B.8. Various Receptors

Receptors for Regulatory Peptides in Epithelial Gut Cancer M . E . F O R G U E - L A F I T T E , M . L A B U R T H E , A . Z W E I B A U M and 595 G . R O S S E L I N

Receptors that Control Acid Secretion in the Isolated Cells from Mammalian Gastric Mucosa 599 R. M A G O U S , M . F . L I G N O N and J . P . B A L I

Microbial Receptors on Human Kidney Epithelia, A Possible Clue in the Pathogenesis of Bacterial Interstitial Nephritis 603 J . E . S C H E R B E R I C H , G . J Ü N G S T , C . G A U H L , W . M O N D O R F and W . S C H O E P P E

SECTION C. MONOCLONAL ANTIBODIES

C . l . Hybridoma Technology

Human-Mouse Hybridoma Cells Producing Antibodies G I U L I A C . B . A S T A L D I , C H A R L E S W I L L E M S , W I L M A A L L E Y N E , 609 P E T E R L A N D S D O R P , W I M P . Z E I J L E M A K E R and M A R I A J A N S S E N

xviii C O N T E N T S

Production of Rat Monoclonal Antibodies with the L O U Rat Non Secreting IR983F Myeloma Cell Line 615 H E R V E B A Z I N

Ant i -Pr , -Gd and Related Cold Agglutinins: Human Monoclonal Antibodies Against Neuraminyl Groups 619 D . R O E L C K E , R. B R O S S M E R and W . E B E R T

Analysis of Carbohydrate Determinants on Membranes of Leukemic Leukocytes using Naturally Occurring Monoclonal Human Antibodies (Cold Agglutinins) 623 B . DÖRKEN, D . R O E L C K E , B . B O H N and W . H U N S T E I N

A Study of Hybridoma Soft Agar Cloning 627 P . H E R I O N , J . -D. F R A N S S E N and J . U R B A I N

Evaluation of Alternative Screening and Selection Procedures in Hybridoma Technology 631 H . J . SCHLÜSENER and F . W . F A L K E N B E R G

Comparison of Screening Techniques Related to the Production of Monoclonal Antibodies 637 J . R O Z I N G , P . J O L I N G , W . V A N E W I J K and C . J . H E S S E

A Semi-Automated Enzyme-Linked Immunosorbent Assay (ELISA) to Screen for Hybridoma Cultures Producing Antibody to Carcinoembryonic Antigen (CEA) 641 C . S. W O O D H O U S E , C . H . J . F O R D and C . E . N E W M A N

Selection of Myeloma Lines Suitable for Hybridoma 645 J . -D. F R A N S S E N , P . H E R I O N and J . U R B A I N

Individual Physical Properties of Mouse Monoclonal Antibodies 649 J . S T E E N S G A A R D

C.2. Definition of Lymphocytes Subsets

Re-Definition of Human T Cells by Monoclonal Antibodies 653 P . C . L . B E V E R L E Y and R. E . C A L L A R D

Biochemical Studies of Human T Cell Markers C O X T E R H O R S T , A N D R E V A N A G T H O V E N , J A N N I E B O R S T , 659 K E N N E T H L E C L A I R and P E T E R S N O W

Mouse Monoclonal Antibodies Defining Functional Human Lymphocyte Subsets 663 E . P . R I E B E R , J . L O H M E Y E R , D . J . S C H E N D E L and G . R I E T H M Ü L L E R

T-Cell Profile and Reactivity I n V i t r o Düring Immune Reconstitution Following Bone Marrow Transplantation 667 J . M . V O S S E N , A . A S T A L D I , H . G . D E B R U I N and L . J . D O O R E N

C O N T E N T S xix

Interaction of Monoclonal Antibodies and Antigen-Specific T-Cell Factor in Regulation of an Immune Response 671 M . J . T A U S S I G

Microfluorometric Analysis of Five Monoclonal Antibodies Directed Against Mouse Thymocytes 675 W . V A N E W I J K , P. W. V A N V L I E T , P . L . V A N SOEST, J . R O Z I N G and G . J . V A N D E N E N G H

Binding Characteristics of a Monoclonal Antibody Against Rabbit Thymocytes 679 J . G . G U I L L E T , P. M A R C H E , J . H O E B E K E and A . D . S T R O S B E R G

Abnormal Helper/Suppressor T-Cell Distribution in Chronic Brucellosis in Humans 683 D . J . R E E N , A . M . E A R L Y , B. L . H O G A N and R. D . T H O R N E S

Monoclonal Antibodies Defining Rabbit Lymphocyte Subpopulations 687 W. D E S M E T , L . B R Y S , M . V A E C K and R. H A M E R S

The Use of Monoclonal Antibodies to Monitor T-Cell Sub-Populations After Human Cardiac Transplantation 691 M . H . Y A C O U B , D . J . R. F R O U D , M . H . A S H R A F , G . C A M P A L A N I , A . T. F O R S Y T H and A . P O M E R A N C E

Monoclonal Anti-B Cell Antibodies: Their Effect on Human Mixed Lymphocyte Reactions 695 C . M . S T E E L , V . V A N H E Y N I N G E N , K . G U Y , B. B. C O H E N , D . L . D E A N E , D . C R I C H T O N and D. H U T C H I N S

Monoclonal Antibodies Against Human Thymocytes and T Lymphocytes W . J . M . T A X , H . W . W I L L E M S , M . D . A . K I B B E L A A R , J . D E G R O O T , 701 P. J . A . C A P E L , R. M . W . D E W A A L , P . R E E K E R S and R. A . P . K O E N E

Monoclonal Antibodies Against Rat Thymocytes 705 J . R O Z I N G , P . J O L I N G , C . J . H E S S E and L . M . B . V A E S S E N

C. 3, Determination of Histocompatibility Antigens

Monoclonal Antibodies Against Human M H C Products P . R I C H I A R D I , F. M A L A V A S I , T. C R E P A L D I , A . O. C A R B O N A R A and 709 R. C E P P E L L I N I

Biochemical Analysis of Human H L A - D R Molecules 713 N . G R O S S , R. S. A C C O L L A , G . C O R T E and S. C A R R E L

Functional Analysis of Human Ia Subsets R. S. A C C O L L A , A . M O R E T T A and J . -C. C E R O T T I N I

719

XX C O N T E N T S

Reactivity of Monoclonal A n t i - H L A Antibodies with Blood Platelets P . M . L A N S D O R P , F. O O S T E R H O F , G . C . B. A S T A L D I , 723 W . P . Z E I J L E M A K E R

Subsets of Human D Locus Products Identified by a Series of Monoclonal Antibodies 729 K . G U Y , V E R O N I C A V A N H E Y N I N G E N , B . B . C O H E N , D . L . D E A N E , D . C R I C H T O N and C . M . S T E E L

C.4. Viral and Bacterial Detection

Monoclonal Antibodies Detecting Epstein-Barr Virus Strain and Group Specific Antigenic Determinants 735 H . H E R B S T , B . G E O R G - F R I E S , N . MÜLLER-LANTZSCH, H . Z U R H A U S E N and D . G . B R A U N

Monoclonal Antibody Against EBV-Associated Transformation Antigen Detected I n V i t r o and I n V i v o 739 D . A . T H O R L E Y - L A W S O N , K . G E I L I N G E R and R. S C H O O L E Y

Production of Monoclonal Antibodies Against Poliomyelitis and Rabies Viruses A . D. M . E . O S T E R H A U S , A . L . V A N W E Z E L , P . V A N D E R M A R E L 743 and G . V A N STEENIS

A Set of Monoclonal Antibodies to Streptococcal Group Polysaccharide Antigens 747 H . H E R B S T , D . L A V A N C H Y and D . G . B R A U N

Hybridoma Antibody Mediated Activation of ß-Galactosidases from Genetically Defective E . C o l i 751 J A N E H E W I T T , R. J U L I A N , S. D U N C A N and P E T E R D . W E S T O N

Characterization of Monoclonal Antibodies to Subunit A of Cholera Toxin 755 A . K U R O S K Y , L . K . D U F F Y and R. M . D E N N E Y

Production of Monoclonal Antibodies Against Mycoplasma Hyorhinis as a Secondary Effect of Immunisation of Mice with Cultured Cell Lines 759 C . V E N N E G O O R , A . A . P O L A K - V O G E L Z A N G and A . H E K M A N

C.5. Detection of Proteins

Monoclonal Antibodies to the Human Leukocyte Interferons and Their Use for Interferon Purification and a Quantitative Interferon Assay 763 T. S T A E H E L I N , B. TAKÄCS, B . D U R R E R , J . S C H M I D T , J . S T O C K E R , V . M I G G I A N O , C. STÄHLI , D . S. H O B B S , H . - F . K U N G and S. P E S T K A

Radioimmunoassay (RIA) of Post-Gamma Globulin Using a Monoclonal Antibody 769 T. S E K I N E , D . J . P E R R Y , S. R A G H E E B and M . D . P O U L I K

C O N T E N T S xxi

Localization of Collagen Type IV in Basement Membranes by Monoclonal Antibodies 773 H . G . F O E L L M E R , J . A . M A D R I , R. T. W Y A T T and H . F U R T H M A Y R

Study of otx Microglobulin (HC Protein) with Monoclonal Antibodies 777 C . V I N C E N T , J . P . R E V I L L A R D , P. B O U I C and J . B R O C H I E R

Anti-Idiotypic Monoclonal Antibodies Against Myasthenie Antibodies 781 R. W . J A M E S , A . - K . L E F V E R T , C . A L L I O D and B. W. F U L P I U S

Quantitation of IgGkappa and IgGlambda in Normal and Pathological Sera 785 M . P R I O R , N . R. L I N G , J . L O W E , S. E V A N S , K . M A Y and R. J E F F E R I S

Use of Monoclonal Antibodies in the Characterisation of Immunoglobulin Classes and Subclasses in Animal Species 789 K . M A Y , S. SENIOR, M . M . G A N I , P . P O R T E R , R. J E F F E R I S and N . L I N G

Monoclonal Antibodies Directed Against a Torpedo Electric Organ Membrane Antigen 793 A . C . V O N G E R S T E N B E R G , U . L 0 V B O R G , C . M . K O C H and E . B O C K

C.6. Identification of Lipids

Monoclonal Antibodies Against Membrane Glycolipids of an Anti-Streptococcal Group A B-Cell Hybridoma 797 R. K . B A L L , H . H E R B S T , C . A . SCHÖNENBERGER, G . R O S E N F E L D E R and D . G . B R A U N

Monoclonal Antibody Secreting Mouse Hybridoma Clones Express Different Sets of Membrane Glycosphingolipids in Different Antigenic Systems 801 G . R O S E N F E L D E R , H . H E R B S T , S. S. A L K A N , C H . H E U S S E R and D . G . B R A U N

Glycosphingolipid Patterns of Established Human Hematopoietic Cell Lines 805 G . R O S E N F E L D E R , A . Z I E G L E R , P. W E R N E T and D. G . B R A U N

Monoclonal Antibodies to Tumor-Associated Glycolipids: Preparation and Application for Suppression of Tumor Growth 809 W . W . Y O U N G , JR , D . L . U R D A L and S. H A K O M O R I

Monoclonal Antibodies Against Human Milkfat Globule Membranes Detecting Differentiation Antigens of the Mammary Gland J . H I L K E N S , F . BUIJS , J . H I L G E R S , P H . H A G E M A N , A . S O N N E N B E R G , 813 U . K O L D O V S K Y , K . K A R A N D E , R. P . V A N H O E V E N , C . F E L T K A M P and J . M . V A N D E R U N

xxii C O N T E N T S

C.7. Determination of Hormones

Monoclonal Antibodies in Clinical Diagnosis — Reagents for Detecting Hormones and Immunoglobulins 817 L . W A N G , C . S. H E X T E R and M . I N B A R

Production of Monoclonal Antibodies to Steroid Hormones 823 Z . E S H H A R , F. K O H E N and H . R. L I N D N E R

Monoclonal Antibodies Against Growth Hormone: Effects on the Hormone Interaction with Specific Cell Surface Receptors 827 L . A . R E T E G U I , P . D E M E Y T S and P. L . M A S S O N

Monoclonal Antibodies Against Human Growth Hormone 833 I. JÖNSDÖTTIR, P . P E R L M A N N and H . P. T. E K R E

Characteristics of Monoclonal Antibodies Against Human Chorionic Gonadotrophin (HCG) 837 A . M . G . B O S C H , W. S T E V E N S , A . S C H U U R S , O. S C H Ö N H E R R and H . R O E L O F S

Sensitive and Specific R I A for Human Chorionic Gonadotropin (hCG) Using Monoclonal Antibodies 843 P. T A N N E R , U . - H . S T E N M A N , M . SEPPÄLÄ and J . SCHRÖDER

The Use of Monoclonal Antibodies Against Prolactin (Prl) in a Solid Phase Enzyme-Immunoassay (EIA) 847 H . V A N D E N B E R G , L . D R I E D O N K S , H . v. G R I N S V E N and A . S C H U U R S

Mouse Monoclonal Antibodies Against Human Prolactine 851 S. K A U L , D . D R A H O V S K Y and A . W A C K E R

Monoclonal Antibodies to Human Prolactin and Chorionic Somatomammotropin 855 J . I V A N Y I and P A T D A V I E S

Monoclonal Thyroglobulin-Autoantibodies as a Tool to Study Autoimmunity G . G U A R N O T T A , R. Q U A R T E Y - P A P A F I O , J . T E M P L E M A N , 861 L . C . P . D E C A R V A L H O and I. M . ROITT

C.8. Determination of Enzymes

Monoclonal Antibodies Against Two Antigens of Clinical Interest C . STÄHLI , P . HÄRING, T. S T A E H E L I N , J . S C H M I D T , R. M U G G L I , 865 P . H U B E R and V . M I G G I A N O

C O N T E N T S xxiii

Monoclonal Antibodies Against Human Acid a-Glucosidase J . H I L K E N S , J . M . T A G E R , F . BUIJS , B. B R O U W E R - K E L D E R , 869 G . M . V A N T H I E N E N , F. P . W . T E G E L A E R S , R. K A L S B E E K and J . H I L G E R S

High-Affinity Monoclonal Autoantibody Used as a Reagent in a R I A for Human Creatine Kinase BB 873 P . U R D A L , P. K I E R U L F and J . H . S T R 0 M M E

C.9. Detection of Cancer Antigens

Five Melanoma-Associated Antigens Identified by Monoclonal Antibodies K . E . H E L L S T R Ö M , I. H E L L S T R Ö M , J . P . B R O W N , M . - Y . Y E H and 877 R. G . W O O D B U R Y

Antigen Expression and Immune Lysis of Human Melanoma Cells: Analysis with Monoclonal Antibodies 883 K . I M A I , A . Y A C H I , A . - K . N G and S. F E R R O N E

Reactions of Monoclonal Antibodies Against Human Melanoma with Different Tissues and Cell Lines 889 P H . H A G E M A N , C . V E N N E G O O R , M . V A N D E R V A L K , J . L A N D E G E N T , A . J O N K E R and L . V A N D E R M I S P E L

Molecular Heterogeneity of a High Molecular Weight Human Melanoma Associated Antigen ( M A A ) Detected by Monoclonal Antibodies 893 K . I M A I , B . S. W I L S O N , G . R U B E R T O , T. N A K A N I S H I , A . Y A C H I and S. F E R R O N E

A Monoclonal Antibody Defining a Burkitt Lymphoma Associated Antigen 899 J . W I E L S , M . F E L L O U S , G . L E N O I R and T. T U R S Z

Preparation and Characterization of Monoclonal Antibodies Against Antigens of Human Renal Adenocarcinoma 903 H . H E S S , U . A L H E I D , S. B I E R M A N N , T. W A K S , J . E . S C H E R B E R I C H , A . W . M O N D O R F , W . S C H O E P P E , T. S E N G E and F. W . F A L K E N B E R G

Monoclonal Antibodies Against Carcinoembryonic Antigen (CEA) 907 A . H E D I N and S. H A M M A R S T R Ö M

Characterisation of Monoclonal Antibodies to Carcinoembryonic Antigen by Antibody Overlay of Polyacrylamide Slab Gels 911 K . A . C H E S T E R and R. H . J . B E G E N T

CIO. Drug Targeting

Phase I Trials of Murine Monoclonal Antibodies to Tumor Associated Antigens: Preliminary Observations R. O. D I L L M A N , R. E . S O B O L and I. R O Y S T O N

915

xxiv

The Potential Use of Monoclonal Antibodies in Drug Targeting G . F. R O W L A N D , R. G . S I M M O N D S , J . R. F. C O R V A L A N , 921 C . H . M A R S D E N , J . R. J O H N S O N , C . S. W O O D H O U S E , C . H . J . F O R D and C . E . N E W M A N

Specific Kill ing of Human and Mouse Tumor Cells by Immunotoxins P . C A S E L L A S , H . E . B L Y T H M A N , J . P. B R O W N , O. G R O S , P . G R O S , 927 K . E . HELLSTRÖM, I. H E L L S T R Ö M , F. K . J A N S E N , P . P O N C E L E T and H . V I D A L

Drug Targeting with Monoclonal Antibodies M . P A G E , J . P . E M O N D , C . G A U T H I E R , C . D U F O U R and L . INNES 933

Analysis of Human Haemopoietic Precursor Cell Antigens Using Astatine Labelled Monoclonal Antibodies 937 G . B R O W N , W . B A T E M A N , J . C O W A N , A . G . F I S H E R , D . T O K S O Z and A . T. M . V A U G H A N

SECTION D. NEW METHODS

The IGS (Immuno Gold Staining) Method Used with Monoclonal Antibodies J . D E M E Y , M . M O E R E M A N S , M . D E W A E L E , G . G E U E N S and 943 M . D E BR A B A N D E R

Enumeration of T Lymphocytes and their Subclasses in Peripheral Blood by Immunogold Staining 949 M . D E W A E L E , J . D E M E Y , M . M O E R E M A N S and B. V A N C A M P

Chromatofocusing: A New High Resolution Method for Protein Fractionation 955 L . SÖDERBERG, T. LÄÄS and D . L O W

The Multiwire Proportional Counter for the Study of Ligand-Binding of Serum Proteins 961 B . J . S C O T T , J . E . B A T E M A N , M . R. H A W K E S W O R T H , D . B U R N E T T and A . R. B R A D W E L L

Identification of Specific Antigens or Antibodies After Electrophoretic Transfer. Application to Measles Virus 965 D . K A R C H E R , A . L O W E N T H A L , H . T H O R M A R and M . N O P P E

A U T H O R I N D E X 969

S U B J E C T I N D E X 977

B . 5 . N e u r o c h e m i c a l Receptors

A S Y M M E T R Y O F A C E T Y L C H O L -INESTERASE A N D A C E T Y L C H O L I N E R E C E P T O R IN I N T A C T S E C R E T O R Y

VESICLES F R O M A D R E N A L M E D U L L A

M . G R A T Z L and H . K R I E G E R - B R A U E R

Department o f Physiology and Chemistry, University o f S a a r l a n d , D - 6 6 5 H o m b u r g / S a a r , F . R . G .

ABSTRACT H i g h l y p u r i f i e d s e c r e t o r y v e s i c l e s from a d r e n a l m e d u l l a , i s o l a t e d by d i f f e r e n t i a l and d e n s i t y g r a d i e n t c e n t r i f u g a t i o n u s i n g i s o t o n i c g r a -d i e n t m a t e r i a l (Percoll™), c o n t a i n a c e t y l c h o l i n e s t e r a s e . The enzyme was l a t e n t i n i s o l a t e d s e c r e t o r y v e s i c l e s i . e . a c e t y l c h o l i n e s t e r a s e was i n a c c e s s i b l e t o added S u b s t r a t e . The enzyme a c t i v i t y became p a t e n t a f t e r a d d i t i o n of d e t e r g e n t or i n h y p o t o n i c media. Hypotonie treatment or s p e c i f i c l y s i s of the v e s i c l e s w i t h Mg2+/ATP i n the presence of a permeant a n i o n r e s u l t e d i n the r e l e a s e of s o l u b l e a c e t y l c h o l i n e s t e r a s e from the v e s i c u l a r c o n t e n t . Membrane-bound enzyme sedimented w i t h the membranes. B i n d i n g of a-bungarotoxin c o u l d o n l y be observed when se­c r e t o r y v e s i c l e s were l y z e d . I t i s c o n c l u d e d , t h a t the a c e t y l c h o l i n e r e c e p t o r as w e l l as the mem­brane-bound form a a c e t y l c h o l i n e s t e r a s e are l o c a l i z e d on the i n n e r s u r f a c e of the s e c r e t o r y v e s i c l e membrane, which becomes the o u t e r s u r f a c e of the c e l l membrane d u r i n g e x o e y t o s i s . C o n c o m i t a n t l y the so­l u b l e form of a c e t y l c h o l i n e s t e r a s e p r e s e n t w i t h i n s e c r e t o r y v e s i c l e s i s r e l e a s e d i n t o the e x t r a c e l l u l a r f l u i d .

KEYWORDS A c e t y l c h o l i n e s t e r a s e ; a c e t y l c h o l i n e r e c e p t o r ; s e c r e t o r y v e s i c l e s ; a d r e n a l m e d u l l a .

INTRODUCTION Duri n g e x o e y t o s i s s e c r e t o r y v e s i c l e c o n t e n t s are r e l e a s e d i n t o the ex­t r a c e l l u l a r f l u i d and t h e i r l i m i t i n g membranes are i n c o r p o r a t e d i n t o the c e l l membrane. In t h i s way the i n s i d e of the s e c r e t o r y v e s i c l e membrane becomes the o u t s i d e of the c e l l membrane. The e x i s t e n c e of c e l l membrane components on the i n n e r s u r f a c e of s e c r e t o r y v e s i c l e membranes would be c o n s i s t e n t w i t h a b i o g e n e t i c r e l a t i o n s h i p between the s e c r e t o r y v e s i c l e and the c e l l membrane. In t h i s r e p o r t we des-c r i b e e v i d e n c e f o r the l o c a t i o n of such c e l l membrane components, the a c e t y l c h o l i n e s t e r a s e and the a c e t y l c h o l i n e r e c e p t o r of the c h r o m a f f i n c e l l , w i t h i n s e c r e t o r y v e s i c l e s .

RESULTS AND DISCUSSION S e c r e t o r y v e s i c l e s from a d r e n a l m e d u l l a , i s o l a t e d by d i f f e r e n t i a l cen-

497

498 NEUROCHEMICAL RECEPTORS

t r i f u g a t i o n , were f u r t h e r p u r i f i e d on a c o n t i n u o u s s e i f g e n e r a t i n g Percoll™ g r a d i e n t . An o s m o l a l i t y of 420 mosmol/kg was m a i n t a i n e d throughout the procedure. In t h i s way h i g h l y p u r i f i e d s e c r e t o r y ve­s i c l e s wereobtained t h a t l e a k e d i n t r a v e s i c u l a r a d r e n a l i n o n l y s l o w l y ( G r a t z l , K r i e g e r - B r a u e r and E k e r d t , 1981). The v e s i c l e s were found t o c o n t a i n a c e t y l c h o l i n e s t e r a s e w i t h a s p e c i ­f i c a c t i v i t y o f about 14 nmoles S u b s t r a t e h y d r o l y z e d per minand mg p r o t e i n ( = t o t a l a c t i v i t y , determined i n the presence of 0.2% T r i t o n X 100). With no d e t e r g e n t added and i n media of an o s m o l a l i t y of 420 mosmol/kg enzyme a c t i v i t y amounted o n l y t o about 4% of the t o t a l ac­t i v i t y ( F i g . 1 ).

£ 0 J i . . - i r -0 100 200 300 400

O s m o l a l i t y ( m o s m o l / k g )

F i g . 1 . Latency of a c e t y l c h o l i n e s t e r a s e of i s o l a ­t e d s e c r e t o r y v e s i c l e s . The v e s i c l e s were i n c u b a t e d f o r 5 min a t room temperature i n sucrose media of d i f f e r e n t o s m o l a l i t i e s . The enzyme a c t i v i t y was determined w i t h o u t d e t e r g e n t added or w i t h 0.2% T r i t o n X 100 p r e s e n t (100%).

R e d u ction of the o s m o l a l i t y r e s u l t s i n an i n c r e a s e of the enzyme a c t i ­v i t y . Below 200 mosmol/kg, an o s m o l a l i t y a t which the v e s i c l e s have d e p l e t e d a l l t h e i r i n t r a v e s i c u l a r a d r e n a l i n ( G r a t z l , 1 9 8 0 ) , no f u r t h e r i n c r e a s e i n enzyme a c t i v i t y c o u l d be d e t e c t e d ( F i g . 1 ) . D uring hypoto-n i c treatment (200 mosmol/kg) of s e c r e t o r y v e s i c l e s about one t h i r d of the t o t a l a c e t y l c h o l i n e s t e r a s e was r e l e a s e d i n t o the s u p e r n a t a n t (Table 1 ) , w h i l e the re m a i n i n g a c t i v i t y sedimented w i t h the membranes. O b v i o u s l y a membrane-bound p o o l o f a c e t y l c h o l i n e s t e r a s e e x i s t s w i t h i n s e c r e t o r y v e s i c l e s a l o n g w i t h the s o l u b l e pool which c o u l d a l s o be r e ­l e a s e d from the v e s i c l e s by 2 mM Mg 2 +/ATP i n the presence of K C l , but not i n the presence of suc r o s e (Table 1 ) . L y s i s of s e c r e t o r y v e s i c l e s as a consequence of an ATP d r i v e n p r o t o n pump i n the presence of a permeant a n i o n has been d e s c r i b e d e a r l i e r (Casey and Co l l e a g u e s , 1 9 7 6 ) . R e c e n t l y we have d e t e c t e d , t h a t the membrane-bound form of a c e t y l c h o ­l i n e s t e r a s e can be d i s t i n g u i s h e d from the s o l u b l e enzyme by i t s e l e c -t r o p h o r e t i c m o b i l i t y ( G r a t z l , K r i e g e r - B r a u e r and E k e r d t , 1981).

ACETYLCHOLINESTERASE IN "INTACT 4' SECRETORY VESICLES 499 Table 1 RELEASE OF ACETYLCHOLINESTERASE FROM

ISOLATED SECRETORY VESICLES

Medium O s m o l a l i t y A c e t y l c h o l i n e s t e r a s e (mosmol/kg) i n Supernatant (%)

Sucrose 420 T71 KCl 4 20 4.7 Sucrose, Mg /ATP 4 20 2.6 K C l , Mg 2 +/ATP 420 23.6 Sucrose 200 32.6

I s o l a t e d s e c r e t o r y v e s i c l e s were i n c u b a t e d f o r 30 min a t 37°C i n 20 mM Mops, pH 7.0, 1 mM EGTA and sucrose or KCl to o b t a i n the f i n a l o s m o l a l i t y as i n d i c a t e d . The c o n c e n t r a t i o n of Mg2 + and ATP was 2 mM. The m i x t u r e s were c e n t r i f u g e d f o r 10 min a t 130000 g a v i n a Beckman Airfuge™ and the enzyme a c t i v i t y was determined i n the s u p e r n a t a n t . T o t a l enzyme a c t i v i t y (=100%) was determined i n the p r e ­sence of 0.2% T r i t o n X 100.

The experiments d e s c r i b e d so f a r show, t h a t s e c r e t o r y v e s i c l e s from a d r e n a l medulla c o n t a i n two types of a c e t y l c h o l i n e s t e r a s e , both se-q u e s t e r e d by the v e s i c u l a r membrane from the s u r r o u n d i n g medium. One type i s membrane-bound, but f a c e s the i n t r a v e s i c u l a r space, the o t h e r type i s p a r t of the v e s i c u l a r c o n t e n t . In the f o l l o w i n g we p r o v i d e e v i d e n c e , t h a t a l s o the a c e t y l c h o l i n e r e c e p t o r i s arranged i n an aSym­m e t r i e way i n the s e c r e t o r y v e s i c l e membrane, namely on the i n n e r s u r ­f a c e . a-bungarotoxin d i d not b i n d t o i n t a c t s e c r e t o r y v e s i c l e s . How-ev e r , when the v e s i c l e s were l y z e d i n the presence of the venom, spe­c i f i c b i n d i n g was observed ( F i g . 2 ) .

c <D O

E "~ < * - B u n g a r o t o x i n ( n M )

F i g . 2 B i n d i n g of [ H] a-bung a r o t o x i n t o s e c r e t o r y v e s i c l e s .

500 NEUROCHEMICAL RECEPTORS

I s o l a t e d s e c r e t o r y v e s i c l e s were l y z e d h y p o t o n i -c a l l y ( f i n a l o s m o l a l i t y 170 mosmol/kg) i n the p r e ­sence of d i f f e r e n t c o n c e n t r a t i o n s of [ 3H] a-bunga­r o t o x i n o r [^H] a-bungarotoxin p l u s a c o n s t a n t amount of unlabelled a -bungarotoxin (5x10~^M). I n -c u b a t i o n was c o n t i n u e d f o r 45 min a t room tempera-t u r e . Membranes were s e p a r a t e d from the medium i n a sucrose Step g r a d i e n t . The r a d i o a c t i v i t y of the m a t e r i a l r e c o v e r e d from the g r a d i e n t was d e t e r ­mined by l i q u i d s c i n t i l l a t i o n c o u n t i n g . S p e c i f i c b i n d i n g was o b t a i n e d from the d i f f e r e n c e between the amount of [ 3H] a-bungarotoxin bound i n the absence and the presence of u n l a b e l l e d a-bungaro­t o x i n .

At the c o n c e n t r a t i o n of 40 nM s p e c i f i c b i n d i n g of a-bungarotoxin was a p p r o x i m a t e l y s a t u r a t e d and the number of b i n d i n g S i t e s was 33 fmole/ mg membrane p r o t e i n . S a t u r a t i o n of a - b u n g a r o t o x i n b i n d i n g S i t e s of c e l l membranes i s o l a t e d from a d r e n a l m e d u l l a was observed a t 4 nM and the number of S i t e s was 190 fmole/mg membrane p r o t e i n (Wilson and K i r s h n e r , 1 9 7 7 ) . Concerning the d i s t r i b u t i o n and the t r a n s p o r t of a c e t y l c h o l i n e s t e r a s e and a c e t y l c h o l i n e r e c e p t o r , i n t e r e s t i n g p a r a l l e l o b s e r v a t i o n s have been made w i t h c h r o m a f f i n c e l l s as w e l l as w i t h myoblasts. Both types of c e l l s i n c u l t u r e r e l e a s e s o l u b l e a c e t y l c h o l i n e s t e r a s e i n t o the me­dium (Rotundo and Famborough,1980a, Rotundo and Famborough,1980, Mizobe and L i v e t t , 1 9 8 0 ) . In m y o b l a s t s , newly formed a c e t y l c h o l i n e s ­t e r a s e and a c e t y l c h o l i n e r e c e p t o r s were found l o c a l i z e d w i t h i n i n t r a -c e l l u l a r o r g a n e l l e s . A l t h o u g h a c e t y l c h o l i n e s t e r a s e i s s e c r e t e d i n a Ca2+-dependent manner by s t i m u l a t e d a d r e n a l g l a n d s , the mechanism of t h i s p r o c e s s remained obscure (Chubb and Smith,1975a,Chubb and Smith, 1975b,Somogyi and C o l l e a g u s , 1 9 7 5 ) . S i n c e i t has been shown now, t h a t a c e t y l c h o l i n e s t e r a s e (as w e l l as a c e t y l c h o l i n e r e c e p t o r ) i s p r e s e n t w i t h i n a d r e n a l m e d u l l a r y s e c r e t o r y v e s i c l e s , an e x o c y t o t i c pathway i s l i k e l y t o o c c u r .

ACKN0WLEDGMENT We thank M r s . M . E l i s f o r e x c e l l e n t t e c h n i c a l a s s i s t a n c e and f o r t y p i n g the m a n u s c r i p t . T h i s work was supported by the S o n d e r f o r s c h u n g s b e r e i c h 38"Membranforschung" of the Deutsche Forschungsgemeinschaft.

REFERENCES Casey,R.P.,D.Njus, G.K.Radda, and P.A.Sehr (1976).Biochem.J.158,583-588. Chubb,I.W., and A.D.Smith (1975a). Proc.R.Soc.Lond.B, 191,24 5-2 61. Chubb,I.W., and A.D.Smith (1975b). Proc.R.Soc.Lond.B, 191,263-269. G r a t z l , M . (1980). T r a n s p o r t of membranes and v e s i c l e c o n t e n t s d u r i n g

e x o e y t o s i s . In T.Bücher, W.Sebald, H.Weiss (Eds), B i o l o g i c a l Che-m i s t r y of O r g a n e l l e F o r m a t i o n , S p r i n g e r V e r l a g , B e r l i n , H e i d e l ­b e r g , pp 165-174.

G r a t z l , M . H . K r i e g e r - B r a u e r , and R.Ekerdt (1981). B i o c h i m . B i o p h y s . A c t a s u b m i t t e d

Mizobe,F., and B . G . L i v e t t (1980). J.Neurochemistry 35, 1469-1473. Rotundo,R.L., and D.M.Famborough (1980). C e l l , 22,583-594. Rotundo,R.L., and D.M.Famborough (1980). C e l l , 22,595-602. Somogyi,P., I.W.Chubb, and A.D.Smith (1975). Proc.R.Soc.Lond.B, 191,

271-283. Wilson,S.P., and N . K i r s h n e r (1977). J.Neurochemistry 28,687-695.

A U T H O R INDEX

A c c o l l a R.S. 713, 719 Acevedo F. 263 A i r o l d i G. 337 A k a h o n a i Y. 93 A l b e c k M.J. 151 A l e x a n d r o v I . 159 A l h e i d U. 903 A l k a n S.S. 801 A l l e g r i n i P.R. 27 A l l e y n e W. 609 A l l i o d C. 781 A l o n z o A. 231 A m i r a n o f f B. 513 Andre C. 493 A s h r a f M.H. 691 A s t a l d i A. 667 A s t a l d i G.C.B. 609, 723 Aubery M. 341, 345

B a f f e t G. 183 B a i l y e s E.M. 33 B a l d u i n i C. 337 B a l i J . P. 599 B a l l R.K. 797 B a l l m e r K. 307 B a r e l A.O. 591 B a r l a t i S. 299, 303 Bateman J . E . 961 Bateman W. 937 B a u d h u i n P. 389 Bause E. 57 B a u t z G. 487 B a z i n H. 615 Begent R.H.J. 911 B e n a s s a y a g C. 369 Berek C. 247

B e r n a r d B. 341, 345 B e v e r l e y P.C.L. 653 B e y r e u t h e r K. 57 B i e r l a i r e F. 547 B i e r m a n S. 903 B i e r m e R. 79 B i j l e n g a R.K.L. 45 B i r c h m e i e r W. 287 B i r s h t e i n B.K. 415 Björck L. 191 Blommesteyn A.C. 271 B l y t h m a n H.E. 927 Bock E. 151, 155, 793 B o e r P. 67 Bohn B. 623 Böhni P. 259 Boneu B. 79 B o n n a f o u s J.C. 147 Bono R. 517 B o r s t J . 659 B o s c h A.M.G. 837 Bosman-Jacobs E.P. 271 Bosman G.J.C.M. 67 B o u i c P. 777 B o u r g e o i s P. 543 B r a d w e l l A.R. 397, 961 B r a u n D.G. 735, 747, 797,

801, 805 B r o c h i e r J . 777 B r o d b e c k U. 105, 109 B r o n C. 133, 255 B r o s s m e r R. 619 B r o u w e r - K e l d e r B. 869 B r o v e l l i A. 337 Brown G. 937 Brown J.P. 877, 927 B r u y n z e e l P.L.B. 163, 167

969

970 AUTHOR INDEX

B r y s L. 687 B u i j s F. 813, 869 B u r g e r M.M. 307 B u r n e t t D. 397, 961 Bygdeman M. 195 Byström B. 195

C a l l a r d R.E. 653 C a m p a l a n i G. 691 C a m p b e l l R. 415 Ca n t a u B. 517 C a n t r e l l J . L . 213 C a p e l P.J.A. 701 Capron A. 179 C a r b o n a r a A.O. 709 C a r r e l S. 713 C a s e l l a s P. 927 Cas s i m a n J . J . 427 C e p p e l i n i R. 709 C e r o t t i n i J.C. 133, 719 Changeux J . P . 351 Chap H.J. 79 C h a t e l a i n P. 381, 509, 531 e h e s t e r K.A. 911 C h r i s t e f f N. 369 C h r i s t i a n s s o n A. 175 C h r i s t o p h e J . 381, 509,

531 C i g e n R. 191 C l a s F. 317 C l a u s e r P. 517 C l e m e t s o n K . J . 359 C l e r c - H o f f m a n n F. 369 C l e v e H. 3 Cochaux P. 579 C o e l J . 551, 555 Cohen B.B. 695, 729 C o l i g a n J . E . 187, 237 C o l o m b i M. 299 C o m o g l i o P.M. 291 C o n s t a n s J . 373 Conway T.P. 201 C o r r a d i n G. 133 C o r t e G. 713 C o r v a l a n J.R.F. 921 Couraud P.O. 493 Cowan J . 937 C r e p a l d i T. 709 C r i c h t o n D. 695, 729 C r i c h t o n R.R. 447

Dahr W. 57, 365 Daum G. 259 D a u t r e v a u x M. 393, 583 D a v i e s P. 855

Daz o r d A. 377 Deane D.L. 695, 729 De B r a b a n d e r M. 943 de B r u i n H.G. 667 de C a r v a l h o L.C.P. 861 de G r o o t J . 701 De Mey J . 943, 949 De Meyts P. 827 De M u l d e r A. 329 de P e t r o G. 303 de R o o i j F.W.M. 271 De Smet W. 687 de Vaux S a i n t C y r C. 159 de Waal R.M.W. 701 De Waele M. 943, 949 D e b u i r e B. 41 D e l a v i e r - K l u t c h k o C. 493 Delorme J . 369 Denney R.M. 755 D e s a i P.R. 213 De s c h e p p e r C. 521 Diamond B. 415 D i e r i c h M.P. 443 D i l l m a n R.O. 915 D i s s o u s C. 179 Dooren L . J . 667 Dornand J . 147 Dörken B. 623 D o r s e t D.L. 171 D o u s t e - B l a z y L. 79 Dr a h o v s k y D. 851 D r i e d o n k s L. 847 D u f f y L.K. 755 D u f o u r C. 933 Dumont I . 333 Dumont J . E . 579 Duncan R.J.S. 751 Dupont C. 513 D u r i e u - T r a u t m a n 0. 493 D u r r e r B. 763 Duthu A. 159

E a r l y A.M. 683 E b e r t W. 619 E k r e H.P.T. 833 Emond J.P. 933 E n e r o t h P. 195, 587 E n g e l A. 171 E n g e r s H. 133 E s s h a r Z. 823 Evans S. 785, 789

F a h r e n k r u g J . 527, 535, 539

F a l k e n b e r g F.W. 631, 903

AUTHOR INDEX 971

F a u l k W.P F e l l o u s M F e l t k a m p F e r r o n e S F e s t e n s t e F e u c h t H F i s c h e r A F o e l l m e r F o n t J . F o o i j M. F o r d C.H F o r g u e - L a F o r m s t e c h F o r s y t h A F r a n s s e n F r o b e r t Y Fröman G. F r o u d D.J Frühling F u l p i u s B F u r t h m a y r

89, 451 899

C. 813 . 883, 893 i n H. 205, 231 E. 405 G. 937

H.G. 773 341 559

J . 641, 921 f i t t e M.E. 595 e r P. 393, 583 .T. 691 J.-D. 627, 645 . 295

263 .R. 691 J . 543 W. 781 H. 49, 773

G u i l l o u z o A. 341 G u s t a f s s o n J.A. 587 Guy K. 695, 729

G a l e t t o G. 291 G a m m e l t o f t S. 527, 535, 539 G a r a v i t o R.M. 171 G a s s e r S. 259 G a t e s F.T. I I I 187 Ga u h l C. 139, 603 G a u t h i e r C. 933 G e e r i n g K. 255 G e i l i n g e r K. 739 G e o r g - F r i e s B. 735 Gerd e s J . 435 G e s e r D. 255 Gespach C. 513 Geuens G. 943 G i b s o n J . 259 G i l l e s C. 521 G i l l o i s M. 79 G i p h a r t M.J. 243 G i r a r d e t M. 255 Ghanem G. 543, 547, 555 G l a s e b r o o k A. 133 Gomez J.M. 571, 575 Goonewardena P. 587 G o s s u i n A. 551, 555, 563 G o u a i l l a r d C. 373 Gräsbeck R. 385, 467 G r a t z l M. 497 G r e a v e s M.F. 227 Gros 0. 927 Gr o s P. 927 G r o s s N. 713 G u a r n o t t a G. 861 G u g u e n - G u i l l o u z o C. 341 G u i l l a u m e M. 547 G u i l l e t J.G. 679

Hadam M.R Haddada H Hageman P Hakomori S. Haldeman A. Ha m e l i n k M.L Hamers R Hamman K Hammarström S. H a n l e y M. 477 Hanson P. 563 Häring P. 865 Hawkesworth M.R. H e d i n A. 907 Heidmann T. Hekman A. H e l e n i u s H e r b s t F H e r b s t H

405 159 813, 889 809 27

167 687

P. 443 907

961

351 759

A. 101 113, 121

735, 747, 797, 801 Hellström I . 877, 927 Hellström K.E. 877, 927 Henry P.E. 555 H e r i o n P. 627, 645 Hess H. 903 Hesse C . J . 637, 705

801 751

817 813, 869 813, 869 15 763 679

C. 63, 71 683 101

71 J . 247

H e u s s e r C. H e w i t t J . H e x t e r C S H i l g e r s J . H i l k e n s H j e r t e n Hobbs D Hoebeke H o e s s l i Hogan B Höglund Hooghe R Hooghe R H o o g h e - P e t e r s E.L. 247 H o r s t W.D. 487 H o r v a t h C. 133 Huang R.T.C. 321 Huber P. 865 H u i j s k e s - H e i n s M.I.E. 459 H u n s t e i n W. 623 H u t c h i n s D. 695

I m a i K. 93, 883, 893 I n b a r M. 817 I n n e s L. 933 I v a n y i J . 855 I v e r s e n L. 477

972

Jägersten C. 175, 251 James R.W. 781 J a n s e n F.K. 927 J a n s s e n M. 609 J a r d S. 517 J a t o n J.C. 45 J e f f e r i s R. 785, 789 J o h a n s s o n K.-E. 175, 251 Jo h n s o n K.R. 921 J o l i n g P. 637, 705 J o n k e r A. 889 J o n s d o t t i r I . 833 Jüngst G. 603

K a i s b e e k R. 869 K a l s h e k e r N. 397 Kampmann L. 275 Karande K. 813 K a r c h e r D. 965 Kauffman H.F. 163 K a u l S. 851 Kawaharada M. 93 Kay M.M.B. 325 Kennes B. 329, 333 K i b b e l a a r M.D.A. 701 K i e r u l f P. 873 K i m b a l l E.S. 187, 237 K i n d t T . J . 187, 237 K i n n e R. 139 K i n n o n C. 221 K i s s o n e r g h i s A.-M. 221 K i t a b g i P. 483 Koch C.M. 793 Ko c h e r H.P. 45 Koene R.A.P. 701 Kohen F. 823 K o l d o v s k y U. 813 K o r d o w i c z M. 57 Kouvonen I . 385, 467 K r e u k n i e t J . 501 K r i e g e r - B r a u e r H. 497 Kung H.-F. 763 K u n i c k i T. 85 K u r o s k y A. 755

Laa s T. 955 L a b u r t h e M. 513, 595 Lande g e n t J . 889 Lang o h r M. 543, 567 La n s d o r p P. 609, 723 L a r i z z a L. 299 Lavanc h y D. 747 Lawson W.J. 125 Leben L. 231 L e C l a i r K. 659

AUTHOR INDEX

Lee C. 477 L e f v e r t A.-K. 781 Le G o f f i c N. 159 L e g r o s F. 543, 547, 551,

555, 563, 567 L e g r o s S. 563 L e j e u n e F . J . 551, 559, 563,

567 L e m a i t r e - C o e l h o I . 419 L e n o i r G. 899 Lepke S. 275 L e s l i e R.G.Q. 431 L e t h i b i c h t h u y Dr. 409 Leyns e B. 459 Le y s e n J . E . 471 L i c h t h a r d t D. 365 L i g n o n M.F. 599 L i m e t J.N. 389, 423, 463 L i n d n e r H.R. 823 L i n g N.R. 785, 789 L i t e a n u A. 563 L i z a n a J . 195 Lögdberg 191 Lohmeyer J . 663 Loos M. 317, 439 L o t s t r a F. 521 L o v b o r g U. 793 Low D. 955 Lowe J . 785, 789 L o w e n t h a l A. 965 L u n d a h l P. 263 Lüning B. 143 Lüscher E. 359 L u s t e n b e r g e r P. 393, 583 L u z i o J .P. 33

MacDonald H. 133 M a d r i J.A. 773 Magous R. 599 M a j u r i R. 467 M a l a v a s i F. 709 Maloy W.L. 187, 237 Mani J.C. 147 Marche P. 679 M a r i e H. 121 Marsden C.H. 921 M a r t i n k o J.M. 237 Marynen P. 427 Masson P.L. 827 May K. 785, 789 M c l n t y r e J.A. 89 Meurs H. 163 Meykens R. 419 M i d d e l k o o p O.P. 313 M i g g i a n o V. 763, 865 M i l s t e i n C.P. 37

AUTHOR INDEX 973

Mimaki T. 487 Mode A. 587 Moeremans M. 943, 949 Moest P. 329 Mondorf A.W. 903 Mondorf W. 139, 603 M o r e i n B. 101 M o r e l l i M. 487 M o r e t t a A. 719 Morgan E.L. 401 M o t t u r a A. 299 M u g g l i R. 865 M u l l e r F. 369 Müller-Lantzsch N. 735 Mu s s e i H.-H. 443

Naim H.Y. 359 N a i r n R. 237 N a k a n i s h i T. 893 Nathenson S.G. 237 N a v a r r o M.A. 571, 575 Neve P. 329, 333 Newby A.C. 33 Newman C.E. 641, 921 Newman R.A. 227 Ng A.-K. 883 Noppe M. 965 N o r s t e d t G. 587 Nuchowicz A. 563 Nunes R.M. 455 Nunez E.A. 369 Nurden A.T. 85

O ' B r i e n R. 487 Octave J.N. 447 Ohashi A. 259 O o s t e r h o f F. 723 O s t e r h a u s A.D.M.E. 74 3 O ' S u l l i v a n M.J. 89 Oswald R.E. 351 O t t P. 105, 109 O t t e s e n B. 527, 535, 539 Owen M.J. 221

Page M. 933 P a l l a v i c i n i G. 337 Pan H. 15 P a p p e r t G. 117 Passow H. 275 P a s t o r M.C. 571, 575 P e r l n a n n P. 833 P e r r y D.J. 769 P e r s s o n H. 97 P e s t k a S. 763 P h i l i p s o n L. 97

P h u t r a k u l S. 263 P i d a r d D. 85 P i n k J.R.L. 71 P i r o P. 333 P o l a k - V o g e l z a n g A.A. 759 Pomerance A. 691 P o n c e l e t P. 927 P o t t e r B . J . 455 P o u l i k M.D. 201, 769 P r i e t o J.M. 455 P r i o r M. 785, 789 Putnam F.W. 41

Q u a r t e y - P a p a f i o R. 861 Q u i n t a r t J . 389

R a a i j m a k e r s J.A.M. 167, 501, 505

R a b b i t s T.H. 37 Ragheeb S. 769 Ramp o l d i E. 299 R e e k e r s P. 701 Reen D. 683 R e t e g u i L.A. 827 R e v i l l a r d J . P. 209, 409, 777 R i c e - E v a n s C. 125 R i c h i a r d i P. 709 R i c k a r t F. 329 R i e b e r E.P. 663 R i e t h m u l l e r G. 663 Riezman H. 259 R o b b e r e c h t P. 381, 509, 531 Robyn C. 567 R o e l c k e D. 365, 619, 623 R o e l o f s H. 837 R o i t t I.M. 861 Roos E. 313 Rosa J .P. 85 Rosenbusch J.P. 171 R o s e n f e l d e r G. 797, 801,

805 R o s s e l i n G. 513, 595 R o t t R. 321 Rowland G.F. 921 R o y s t o n I . 915 R o z i n g J . 637, 675, 705 R u b e r t o G. 893 R u d i o f f V. 113, 121 Rungger-Brändle E. 63

S a a l J.G. 405 Saez J.M. 377 Samarut C. 409 S a n d b e r g B. 477

974 AU'VHOK IN DFX

S a n d e r s o n A.R. 205 Savu L. 369 S c h a t z G. 259 S c h e i n e r 0. 443 S c h e n d e l D.J. 663 S c h e r b e r i c h J . E . 139, 603,

903 Schlüsener H.J. 631 Schmid M. 133 Sc h m i d t J . 763, 865 S c h m i d t W. 205, 231 S c h m i t t M. 443 Schmutz A. 493 Schneck J . 415 S c h n e i d e r Y . - J . 423, 447, 463 Schoeppe W. 139, 603, 903 Schönenberger C.A. 797 Schönherr 0. 837 S c h o o l e y R. 739 Schorlemmer H.U. 439 S c h r e i b e r A.B. 493 S c h r e i e r M. 71 Schröder J . 843 S c h u b e r t D. 117, 121 S c h u u r s A. 837, 847 S c o t t B . J . 961 S e k i n e T. 769 Senge T. 903 Seppälä M. 843 S i a d a k A.W. 247 S i d d l e K. 33 S i e P. 79 S i g r i s t H. 27 Simmonds R.G. 921 Simons K. 101 S l a c h m u y l d e r - O t t e C. 463 S m i t S. 459 S m i t h R.H. 125 Snow P. 659 S o b o l R.E. 915 Söderberg L. 955 S o l e r J . 571, 575 Sonnenb e r g A. 813 S o u p a r t A. 563 S p r i n g e r G.F. 213, 217 S t a e h e l i n T. 763, 865 Stähli C. 763, 865 S t a u n - O l s e n P. 527, 535,

539 S t e e l C.M. 695, 729 S t e e n s g a a r d J . 649 S t e e n h o u t P. 547 S t e i n H. 435 S t e i n - P a r v e E.P. 67 Stenman U.-H. 843 S t e v e n s W. 837 S t o c k e r J . 763

Stromme J.H. 873 S t r o s b e r g A.D. 493, 679 Su n d q u i s t B. 101 S u t h e r l a n d D.R. 227

Tager J.M. 869 Takacs B. 763 Tanner P. 843 Tarone G. 291 T a u s s i g M.J. 671 Tax W.J.M. 701 T e g e l a e r s F.P.W. 869 Tegtmeyer H. 213, 217 Temmerman A. 555, 559 Templeman J . 861 T e n c h i n i M. 299 T e r h o r s t C. 659 T e r p s t r a G.K. 501, 505 Thoman M.L. 401 Thorley-Lawson D.A. 739 Thormar H. 965 Thornes R.D. 683 Toksoz D. 937 Tr o u e t A. 423, 447, 463 Tursz T. 899

Uehara H. 237 U r b a i n J . 627, 645 U r d a l D.L. 809 U r d a l P. 873

Vaeck M. 687 Vaerman J.P. 419, 423 Vaessen L.M.B. 705 V a h e r i A. 303 V a l l e t t e G. 369 v§n Agthoven A. 6^9 Van Camp B. 949 van de P a v e r t I.V. 313 van de R i j n J.M. 813 van den Be r g H. 847 van den Be r g J.W.O. 271 van den Bogaard W. 163, 167 van den Engh G.J. 675 van d er F e l t z M.J.M. 243 van d er H o f S t a d t G.A.J.M. 75 van d er Ko r p u t J.A.G.M. 243 van d e r M a r e l P. 743 van d er M i s p e l L. 889 van d er V a l k M. 889 van de R i j n J.M. 813 van E w i j k W. 637, 675 Van Gompel P. 471 van Heyningen V. 695, 729

AUTHOR INDEX 975

Y a c h i A. 93, 883, 393 Yacoub M.H. 691 Yamamura H.I. 487 Yamamura S.H. 487 Yao K. 15 Yeh C.-J.G. 451 Yen M.-Y. 877 Young W.W. J r . 809

Z a h l e r P. 27, 267 Z a k i L. 279 Zeeuws R. 591 Z e i j l e m a k e r W.P. 609, 723 Z i e g l e r A. 805 Z u i n g h e d a u J . 159 z u r Hausen H. 735 Zweibaum A. 595

Vercammen-Grandjean A. 559 V e r h e s t A. 563 V e r s e e V. 591 Verwey H. 129 V i a u M. 373 V i d a l H. 927 V i e r e n d e e l s G. 521 V i n c e n t C. 209, 777 von G e r s t e n b e r g A.C. 155, 793 von G r i n s v e n H. 847 V o s s e n J.M. 667 V r a y B. 493

Wacker A. 851 Wada T. 93 W a e l b r o e c k M. 381, 509 Waks T. 903 Wang L. 817 W a r r i n e r G.A. 89 Watson S. 477 Watt W.B. 125 W e i g l e W.O. 401 Wernet P. 805 Wes t b r o e k D.L. 243 Weston P.D. 751 W i e l s J . 899 W i e s l a n d e r A. 175 W i k l u n d B. 143 W i l k i n s o n J.M. 187 W i l l e m s C. 609 W i l l e m s H.W. 701 W i l s o n B.S. 893 W o l f M. 267 Wood P.G. 283 Woodbury R.G. 877 Woodhouse C S . 641, 921 W r o b l e w s k i H. 183 Wyatt R.T. 773

van Hoeven R.P. 813 Van Leuven F. 427 Van Rymenant M.E. 75 Van Sande J . 579 van S o e s t P.L. 675 van S t e e n i s G. 743 van S t e v e n i n c k J . 129 van T h i e n e n C M . 869 Van Tieghem N. 559 van V l i e t P.W. 675 Van Wezel A.L. 743 Vand e r h a e g h e n J . J . 521 V a r t i o T. 303 V a s s a l l i P. 71 Vaughan A.T.M. 937 Vennegoor C 759, 889 V e r b a n c k P. 521 V e r b i s t A. 543

S U B J E C T INDEX

R e c e p t o r s and Membrane P r o t e i n s have been l i s t e d u n d e r t h e i r s p e c i f i c names.

A c e t y l c h o l i n e 599 A c e t y l c h o l i n e s t e r a s e 105, 109, 497 A c e t y l c h o l i n e r e c e p t o r 497, 781 A - c h a i n 927 A c h o l e p l a s m a l a i d l a w i i 15, 175, 251 A c i d a l p h a - g l u c o s i d a s e 869 A c i d s e c r e t i o n 599 ACTH 377 A c t i n 295 A c t i v a t e d T c e l l s 729 A c u t e l e u k e m i a 623 ADCC 883 A d e n o v i r u s g l y c o p r o t e i n 97 A d e n y l a t e c y c l a s e 381, 509, 531,

579 A d e n y l a t e c y c l a s e i n h i b i t i o n 517 A d h e s i v e n e s s 295 A d j u v a n t 401 A d r e n a l m e d u l l a 497 A f f i n i t y c h r o m a t o g r a p h y 591, 763 A g e i n g 325, 329, 333,. 341 A g g l u t i n a t i o n 747 A i r w a y s - o f t h e r a t 501, 505 A l p h a - a d r e n o c e p t o r 505 A l p h a - f o e t o p r o t e i n 369, 591 A l p h a - g l u c o s i d a s e 869 A l p h a ^ - m i c r o g l o b u l i n 777 A l p h a - m a c r o g l o b u l i n 427 Alpha-MSH 551, 555, 559, 563 Alpha-MSH r e c e p t i v i t y 551, 563 A l p h a - t i s s u e a n t i g e n ( a l p h a TA) 93 Amino a c i d s e q uence 41, 57, 187,

237

Ampholyte d i s p l a c e m e n t c h r o m a t o ­g r a p h y 955

A n a l y t i c a l z o n a l c e n t r i f u g a t i o n 649

A n g i o t e n s i n r e c e p t o r s 517 A n i m a l ' I m m u n o g l o b u l i n s ' 789 A n i o n t r a n s p o r t 275 A n i o n t r a n s p o r t and a r g i n i n e s p e c i f i c

r e a g e n t s 279 A n i o n - t r a n s p o r t p r o t e i n 113 A n t i - b e t a ^ M-goat serum 201 A n t i Na (sodium) 255 A n t i b o d i e s t o c e l l s u r f a c e 291 A n t i b o d y b i n d i n g t e s t 759 A n t i b o d y d e t e c t i o n 627 A n t i b o d y f i x a t i o n 159 A n t i b o d y f r a g m e n t s 419 A n t i b o d y h e t e r o g e n e i t y 649 A n t i b o d y r e s p o n s e 671 A n t i CEA 641, 921 A n t i g e n d e n s i t y 883 A n t i g e n s - s u r f a c e 247 A n t i g e n s p e c i f i c f a c t o r 671 A n t i g e n i c a r c h i t e c t u r e 251 A n t i g e n i c c r o s s - r e a c t i v i t y o f TPA

143 A n t i - g l u c o c o r t i c o i d 393 Ant i - H C G 817 A n t i h l g G 817 A n t i - i d i o t y p e s 563 A n t i - 5 ' n u c l e o t i d a s e serum 147 A n t i - r e c e p t o r a n t i b o d i e s 419 Anti-RHD a n t i b o d i e s 609

977

978 SUBJECT INDEX

A n t i s e r u m a g a i n s t C3R 435 A n t i - T 4 817 A p l a s t i c anemia 667 A r y l i s o t h i o c y a n a t e s 27 A s s o c i a t i v e r e c o g n i t i o n 231 A s t a t i n e 937 ATP d e p l e t i o n 109 A t t a c h m e n t f a c t o r s 287 A u t o - a n t i b o d y 325, 365 A u t o r a d i o g r a p h y 911

B a c t e r i a l membrane r e c e p t o r s 603 Bands 1 and 2 p r o t e i n 121 Band 3 p r o t e i n 113, 117, 275,

279 B a s a l - l a t e r a l membranes 139 Basement membrane c o l l a g e n 773 B e n z o d i a z e p i n e 487 B e t a - a d r e n e r g i c b i n d i n g S i t e s 167 B e t a a d r e n e r g i c c a t e c h o l a m i n e r e c e p ­

t o r s 493 B e t a a d r e n e r g i c r e c e p t o r 163 B e t a a d r e n o c e p t o r 501, 505 B e t a ^ a d r e n o c e p t o r 505 17 b e t a c a r b o x a m i d e S t e r o i d 393 B e t a - g a l a c t o s i d a s e 751 B e t a I H - r e c e p t o r s 443 B e t a - m i c r o g l o b u l i n 187, 191,

195, 201, 205, 209, 243 B e t a ^ - m i c r o g l o b u l i n - b i n d i n g 191 B e t a - t h r o m b o g l o b u l i n 865 B i e x p o n e n t i a l d i s s o c i a t i o n k i n e -

t i c s 539 B i l e a c i d s 271 B i o s y n t h e t i c hGH 833 B i o t i n - a v i d i n 809 B l o o d g roup human - M and N s p e c i -

f i c i t i e s i n g u i n e a p i g 213 B l o o d g roup human M-, N-, T-, TN-

s p e c i f i c i t i e s i n e x t r a c t s o f L-10 C a r c i n o m a 213

B l o o d g r o u p s 49, 57 B l o o d p l a t e l e t s 723 Bone marrow t r a n s p l a n t a t i o n 667 Boyden Chamber 201 B r a i n s p e c i f i c p r o t e i n s 151 B r u c e l l o s i s 683 B r u s h b o r d e r 271 B r u s h b o r d e r v e s i c l e s 139 B u r k i t t ' s lymphoma 739

C-mu gene 37 C - $ J e l t a gene 37 CA - d e p e n d e n t c o m p l e x e s 85 C A - i n d u c e d K l e a k 283 c ALL a n t i g e n 227

C a p p i n g 405 C a r b o h y d r a t e d e t e r m i n a n t s 623 C a r b o h y d r a t e r e c e p t o r s 389 C a r c i n o m a a s s o c i a t e d T h o m s e n - F r i e d e n -

r e i c h (T) s p e c i f i c i t y 213 C a r c i n o e m b r y o n i c a n t i g e n 93, 6 4 1 ,

907, 911, 933 C a r d i a c t r a n s p l a n t 691 C a t h e p s i n s 397 CEA see c a r c i n o e m b r y o n i c a n t i g e n C E A - l i k e p r o t e i n 195 C e l l a d h e s i o n 291 C e l l f u s i o n e f f i c i e n c y 645 C e l l r n e d i a t e d c y t o t o x i c i t y 231 C e l l s o r t e r 675 C e l l s u r f a c e components 313, 345 C e l l u l a r t r a n s f o r m a t i o n 303 C e n t r a l n e r v o u s System 521 c GMP 333 C h e m i l u n i n e s c e n c e immunoassay 823 C h e m o t a x i s 201 C h i c k b r a i n 151 C h o l a t e e x t r a c t 263 C h o l e c y s t o k i n i n 521 C h o l e r a t o x i n 755 C h o r i o n i c somatomammotropin 855 C h r o m a t o f o c u s s i n g 777, 955 C h r o n i c l y m p h o c y t i c l e u k e m i a 915 CL 2L8, 872 487 C l a s s s w i t c h 37 C o l d a g g l u t i n i n s 619, 623 C o l d a g g l u t i n i n d i s e a s e 365 C o l l o i d a l g o l d 943 C o m p e t i t i o n c u r v e s 555 Complement ( C l , C2, C A ) 317 Complement C 1 439 Complement dependent l y s i s 883 Complement (C3) r e c e p t o r s (C3R)

435 Complement r e c e p t o r s 443 C o n c a n a v a l i n - A r e c e p t o r 147 C o n d i t i o n e d media 631 C^ r e c e p t o r s 443 C r e a t i n e k i n a s e BB i s o e n z y m e 873 C r o s s e d i m m u n o e l e c t r o p h o r e s i s 183,

251 C r o s s l i n k i n g 27 C r o s s r e a c t i o n s 191 C r o s s r e a c t i v i t y 789 C r y s t a l l o g r a p h i c a n a l y s e s 171 C y c l i c a d e n o s i n e monophosphate 571 C y c l i c AMP 513, 579 C y c l i c n u c l e o t i d e s 575 Cytochrome C 259 C y t o p h i i i c IgG 405 C y t o s k e l e t a l p r o t e i n s 287 C y t o t o x i c i t y 937

SUBJECT INDEX 979

Daunomycin 933 D e n s i t y d e p e n d e n t g r o w t h c o n t r o l

307 D e t e r g e n t s 15, 105, 171, 183 D - g l u c o s e t r a n s p o r t 263 D i f f e r e n t i a t i o n 67, 345, 4 5 1 ,

559, 659 D i f f e r e n t i a t i o n a n t i g e n s 813 D i f f e r e n t i a t i o n by s p e c i f i c a n t i ­

body 743 D i f f e r e n t i a t i o n m a r k e r s 805 D i m e t h y l d i s u l p h i d e 125 D i t h i o e r y t h r i t o l 263 o, o • - d i t y r o s i n e 129 DLA 243 DNA s e q u e n c e 41 Dopamine a g o n i s t 471 Dopamine a n t a g o n i s t 471 Dopamine r e c e p t o r b i n d i n g 471 Down r e g u l a t i o n o f r e c e p t o r s 539 Drug t a r g e t i n g 933

EBV 739, 899 EBV - e n v e l o p e p r o t e i n s 735

- s p e c i f i c m o n o c l o n a l a n t i b o ­d i e s 735

E l e c t r o n d i f f r a c t i o n 171 E l e c t r o n m i c r o s c o p y 171 E l e c t r o p h o r e s i s 911 E l e d o i s i n 477 E l i s a 637, 641, 773 Embryo 345 E n d o c y t o s i s 459 E n d o t h e l i a l c e l l s 609 E n t e r o h e p a t i c c i r c u l a t i o n 271 Enzyme immunoassay 837, 847 E p i t h e l i a l c e l l s 595 E p i t h e l i u m 513 E p i t o p e c o n s e r v a t i o n 789 E p s t e i n - B a r r v i r u s see EBV E r y t h r o c y t e a g i n g 337 E r y t h r o c y t e membrane r e c o n s t i t u t i o n

105 E r y t h r o c y t e membranes 109, 113,

117, 121, 337, 365, 471 E r y t h r o c y t e s 15, 125, 275, 279 E s c h e r i c h i a c o l i 603 E s t r o g e n b i n d i n g 591 E s t r o g e n s 369 E v o l u t i o n 191 Exchange 205 E x t r a c e l l u l a r m a t r i x 307 E x t r a n e u r a l r e c e p t o r s 483

Fa c s 637

Fe f r a g m e n t s 401 Fe r e c e p t o r 439 Fe gamma r e c e p t o r 409 Fe r e c e p t o r s 385 Female g e n i t a l t r a c t 527 F e m i n i z i n g f a c t o r 587 F i b r o b l a s t s 459 F i b r o n e c t i n 291, 299 F i b r o n e c t i n d e g r a d a t i o n p r o d u e t s

303 F l o w m i c r o f l u o r o m e t r y 687 F l u o r e s c e n c e a c t i v a t e d c e l l s o r t e r

687 F l u o r o m e t r y 679 F o c a l c o n t a c t s 287 F o r s s m a n - l i k e a n t i g e n 159 F r e e f l o w e l e c t r o p h o r e s i s 139 F r i e n d c e l l 67

G a n g l i o s i d e s 797 G a n g l i o t r i o s y l c e r a m i d e 809 Gardos e f f e c t 283 G a s t r i c mueosa 599 G a s t r i n 599 G l u c o c o r t i c o i d 567 G l u c o c o r t i c o i d r e c e p t o r 393, 583 G l y c o c a l i c i n 359 G l y c o g e n 595 G l y c o p h o r i n 49 G l y c o p r o t e i n 205 KD 435 G l y c o p r o t e i n s 57, 67, 71, 89,

221, 325, 365, 659 G l y c o s p h i n g o l i p i d s 801, 805 G o n a d o t r o p i n s 567 Gram-neg. b a c t e r i a 317 G r a n u l o c y t e s 295 G r a v e s ' d i s e a s e 571 G r o s s V i r u s l e u k a e m i a 231 Group N a n t i g e n a n t i - a s i a l o -

g a n g l i o s i d e 217 Growth 595 Growth f a c t o r s 307 Growth hormone 827 Growth p h a s e s : h o r i z o n t a l and v e r t i -

c a l 551 Growth r e g u l a t i o n 307 GTP 513 G u a n i n e n u c l e o t i d e s 381 G u i n e a p i g L-10 h e p a t o c a r c i n o m a 213 Gut 513, 595

r e s t r i c t i o n 231 H a e m o p h i l u s i n f l u e n z a e 501 H a i r y - c e l l l e u k a e m i a 75 Hamster 159

980 SUBJECT INDEX

H a p t o g l o b i n 397 HCG 837 H 2 DIDS 275 H - D I - h y d r o - A l p r e n o l o l 163 H e a r t 381, 531 Heavy c h a i n C - r e g i o n genes 37 H e i n z body anaemia 125 H e m a g g l u t i n i n 321 Herne hemopexin 463 Herne r e c e p t o r 467 Hemin 467 H e p a t o c y t e membrane b i n d i n g 547 H e p a t o c y t e s 313, 389, 419, 423,

455, 463, 535 Hexose u p t a k e 345 H i g h - a f f i n i t y a u t o a n t i b o d y 873 H i s t a m i n e 599 H i s t o c o m p a t i b i l i t y 187 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 205,

209, 221, 237 H i s t r i o n i c o t o x i n 351 HLA - A, B, C 709 HLA a n t i g e n s 723 HLA-D 729 HLA-DR 227, 713, 729 Homozygous t y p i n g c e l l s 729 Hormone immunoassay 855 Hormone r e c e p t o r s 509 Hormone r e g u l a t i o n 575 Human b i o s y n t h e t i c i n s u l i n 547 Human c h o r i o n i c g o n a d o t r o p i n (HCG)

843 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) 609 Human e r y t h r o c y t e g h o s t s 267 Human e r y t h r o c y t e membrane 263 Human g r o w t h hormones 833 Human g r o w t h hormones s u b f r a c t i o n s

587 Human h e m a t o p o i e t i c c e l l l i n e s 805 Human IgD 37 Human k i d n e y 139, 773 Human l i v e r c e l l s 341 Human l y m p h o c y t e s 409 Human melanoma a s s o c i a t e d a n t i g e n

( H i g h M o l e c u l a r W e i g h t ) 893 Human melanoma c e l l s 555, 559,

883 Human melanomas 559, 563, 877, 927 Human mouse h y b r i d o m a c e l l s 609 Human p r o l a c t i n 851, 855 Human r e n a l a d e n o c a r c i n o m a 903 Human s u p e r f i c i a l s p r e a d i n g m e l a ­

noma 551 Human-T-lymphocytes 405 Human t r i a l s 915 H y a l u r o n i c a c i d 341

Hybridoma 247, 615, 627, 809, 855, 899, 921

Hybr i d o m a a n t i b o d y 907 Hyb r i d o m a g r o w t h p a t t e r n 627, 645 Hybr i d o m a t e c h n o l o g y 631 H y d r o p h o b i e c h r o m a t o g r a p h y 15,

359 H y d r o p h o b i e c o n s t a n t domain 45 H y d r o p h o b i e i n t e r a c t i o n s 105 H y p o c h o l e s t e r o l a e m i a 125 H y p o p h y s i s 521

IA a n t i g e n s 719 IgA r e c e p t o r 419 I g D 45 I g Fe p o r t i o n 401 I g G 3 809 IgG immune c o m p l e x e s 649 I g M 45, 809 Immune s u r v e i l l a n c e 231 Immunochemical c h a r a c t e r i z a t i o n

o f TPA 143 I m m u n o c y t o c h e m i s t r y 943 I m m u n o c y t o c h e m i s t r y a u t o r a d i o g r a p h y

555 I m m u n o d i a g n o s i s 877 I m m u n o f l u o r e s c e n c e 773 I m m u n o g l o b u l i n q u a n t i t a t i o n 785 I m m u n o g l o b u l i n r e c e p t o r s 431 I m m u n o g l o b u l i n s 4 1 , 789 Immunogold s t a i n i n g 943, 949 I m m u n o h i s t o c h e r n i c a l - a u t o r a d i o g r a -

p h i c v i s u a l i z a t i o n 547 I m m u n o h i s t o c h e m i s t r y 889 I m m u n o h i s t o l o g y 773 I m m u n o l o g i c a l m o n i t o r i n g 691 Immunoperoxidase t e c h n i q u e 723 I m m u n o p r e c i p i t a t i o n 155, 735,

793 I m m u n o r e g u l a t i o n 671 I m m u n o r e g u l a t o r y d r u g s 683 Immunotherapy 877 Immunotoxins 927 I n f e c t i o u s m o n o n u c l e o s i s 739 I n f e r t i l i t y 195 I n f l u e n z a v i r u s - i n d u c e d f u s i o n 321 I n h i b i t i o n b i n d i n g a s s a y s 893 I n s i d e - o u t v e s i c l e s 267 I n s u l i n 595 I n s u l i n m e t a b o l i s m 543 I n s u l i n p o r c i n e 547 I n s u l i n r e c e p t o r s 535 I n t e r f e r o n 763 I n t e r f e r o n t r e a t m e n t 883 I n t e r n a l i z a t i o n 547 I n t e r s p e c i e s c r o s s r e a c t i v i t y 255

SUBJECT INDEX 981

I n t e s t i n a l t r a n s p o r t 271 I n t r i n s i c f a c t o r r e c e p t o r 377 I n v i t r o S t i m u l a t i o n 609 I n v i t r o t r a n s l a t i o n 97 I n v i v o g r o w t h 645 I n v i v o s c i n t i g r a p h i c s t u d i e s 543 I o n C h a n n e l 351 I o n p a i r 15 I r o n 459, 467 I r o n u p t a k e 455 I s o e l e c t r i c p o i n t 955 I s o l a t e d c e l l s 599

K-ATPase a n t i b o d i e s 255 K + f l u x e s 329

L a b e l l i n g 27 L e c t i n s 307, 333 Leukaemia 227, 451 L i g a n d b i n d i n g s i t e s 373 L i p o p o l y s a c c h a r i d e s 317 Liposome a g g l u t i n a t i o n 79 Li p o s o m e s 385 L i v e r 539 L i v e r a d e n y l a t e c y c l a s e 517 L i v e r m i c r o s o m e s 587 LH 837 L o c a l a n e s t h e t i c s 351 Lou r a t 615 Lymphocyte 71 Lymphocyte a c t i v a t i o n 329, 333 Lymphocyte c y t o s k e l e t o n 63 Lymphocyte S t i m u l a t i o n 147 Lymphocyte s u b p o p u l a t i o n s 663,

949 Lymphocytes 159, 163, 167, 221,

401, 943 Lymphosarcoma 313 Lys o s o m a l enzymes 439 L y s o m o t r o p i c a g e n t s 447, 463

Macro CK-BB 873 Macrophages 389, 431 Majo r h i s t o c o m p a t i b i l i t y 231 Maj o r h i s t o c o m p a t i b i l i t y c omplex

729 Mammary g l a n d s 813 M a n t i g e n 57 Melanoma 889, 915 Membrane a n t i g e n s 71, 155, 679,

793 Membrane a s s o c i a t e d t i s s u l a r a u t o -

a n t i g e n (MTA) 93 Membrane asymmetry 251 Membrane f u s i o n 267

Membrane g l y c o p r o t e i n s 93, 337 Membrane l y s i s 603 Membrane m o l e c u l e 237 Membrane p r o t e i n C l u s t e r s 337 Membrane p r o t e i n b i o s y n t h e s i s 221 Membrane p r o t e i n s o l u b i l i z a t i o n 171 Membrane p r o t e i n s 15, 27, 39, 133

317 Membrane r e c o n s t i t u t i o n 79 Membranes 175 Messenger RNA 97 MHC 243 M i c r o b i a l a d h e r e n c e 603 M i c r o s c o p y - l i g h t 949

- l i g h t and e l e c -t r o n 943

M i l k f a t g l o b u l e s 813 M i t o c h o n d r i a 259 M i t o g e n i c l e c t i n s 147 M i x e d l y m p h o c y t e r e a c t i o n s 695,

719 MNSS System 471 M o l e c u l a r h e t e r o g e n e i t y 893 M o l e c u l a r w e i g h t o f C3R 435 M o n o c l o n a l a n t i b o d i e s t o human IgG

649 M o n o c l o n a l a n t i b o d i e s t o e s t r o g e n s

823 M o n o c l o n a l a n t i b o d i e s t o human l e u k o -

c y t e i n t e r f e r o n s 763 M o n o c l o n a l a n t i b o d i e s 33, 227,

365, 427, 615, 619, 637, 641, 653, 659, 663, 667, 671, 675, 679, 687, 691, 695, 701, 705, 709, 719, 729, 737, 743, 747, 751, 755, 759, 769, 773, 777, 781, 789, 793, 797, 801, 813, 827, 833, 837, 843, 847, 8 5 1 , 855, 865, 869, 877, 883, 889, 893, 903, 911, 927, 933, 943, 949

M o n o c l o n a l a n t i - H L A a n t i b o d i e s 723 M o n o c l o n a l a u t o a n t i b o d i e s 861 M o n o c l o n a l human a n t i b o d i e s 623 Monocytes 201 M o n o m e r / d i m e r / t e t r a m e r a s s o c i a t i o n

e q u i l i b r i u m 117 Mouse h y b r i d o m a 801 M g a n t i g e n 57 M u l t i p l e s c l e r o s i s 965 M u l t i w i r e p r o p o r t i o n a l c o u n t e r 961 M u r i n e m o n o c l o n a l a n t i b o d i e s 915 M u r i n e T c e l l c l o n e s 133 M u s c a r i n i c r e c e p t o r s 381 M u t a t i o n a l h y p o t h e s i s o f C l a r k 551 M y a s t h e n i a g r a v i s 781 Mycoplasma i d e n t i f i c a t i o n 759 Myeloma c e l l s 645

982 SUBJECT INDEX

Myometrium 527

N - a c e t y l n e u r a m i n i c a c i d 619 N a + , K + A T P a s e 139, 329 N a t u r a l k i l l e r a c t i v i t y 653 N a t u r a l k i l l e r c e l l h e t e r o g e n e i t y

663 N e g a t i v e c o o p e r a t i v e i n t e r a c t i o n s

539 Neo a n t i g e n i c d e t e r m i n a n t s 427 N e u t r a l g l y c o s p h i n g o l i p i d s 797 N e u r a l r e c e p t o r s 483 N e u r a m i n i d a s e 321, 619 N e u r a m i n i d a s e - r e s i s t a n t 75 Neuron 247 N e u r o p e p t i d e s 521, 535 N e u r o t e n s i n - r e c e p t o r i n t e r a c t i o n s

483 N e u t r a l i z a t i o n t e s t 559 N o n - e s t e r i f i e d f a t t y a c i d s 369 N u c l e a r m a g n e t i c r e s o n a n c e 49 5 ' N u c l e o t i d a s e 33, 147

Ozone 129

P a r a i n f l u e n z a - 3 - v i r u s 101 P a r a m y x o v i r u s - i n d u c e d f u s i o n 321 P a t h o l o g i c a l I m m u n o g l o b u l i n s 571,

PBE 955 P e p t i d e hormones 477, 535 P e r i t o n e a l macrophages 439 P h a g o c y t o s ^ s 295 P h a r m a l y t e 955 P h e n c y l l i d i n e 351 P h o s p h o d i e s t e r a s e 513 P h o s p h o l i p a s e 2 6 7 P h o s p h o l i p i d l i p o s o m e 263 P h o s p h o l i p i d m o n o l a y e r s 121 P h o s p h o l i p i d s p e c t r i n i n t e r a c t i o n s

121 P h o s p h o l i p i d s 105 P h o t o a f f i n i t y l a b e l l i n g 351 P h y s a l a e m i n 477 P i g m e n t e d and u n p i g m e n t e d m e l a n o c y t e s

563 P i t u i t a r y p r o t e i n hormones 855 Pl a s m a hormone l e v e l s 567 P l a s m a membrane 33 P l a s m a membrane p r o t e i n s 291 P l a s m i n o g e n a c t i v a t o r 299 P l a t e l e t a g g r e g a t i o n 85 P l a t e l e t g l y c o p r o t e i n 79 P l a t e l e t membrane g l y c o p r o t e i n I B

359

P l a t e l e t membrane g l y c o p r o t e i n s I I b and l i l a 85

P o l i o m y e l i t i s v i r u s 743 P o l y b u f f e r 955 P o l y c l o n a l a n t i b o d y 769 P o l y m e r i e IgA 423 P o l y p e p t i d e s 18 and 38 KD 435 Pompe's d i s e a s e 869 P o r t a l r o u t e i n f u s i o n 547 P o s t gamma g l o b u l i n 769 P r e - and p o s t m e n o p a u s a l women 5 6 ^ P R - a n t i g e n s 365 P r e c u r s o r c e l l s 937 P r o l a c t i n l i v e r r e c e p t o r s 587 P r e g n a n c y 837 P r e p a r a t i v e a g a r o s e - s u s p e n s i o n e l e c t

p h o r e s i s 183 P r o l a c t i n 567, 847 P r o s t a t i c a c i d P h o s p h a t a s e 865 P r o t e a s e s 259, 397 P r o t e a s e i n h i b i t o r s 397 P r o t e i n A 1 5 9

P r o t e i n a c y l a t i o n 113 P r o t e i n c r o s s - l i n k i n g 129 P r o t e i n f r a c t i o n a t i o n 155, 955 P r o t e i n - l i p i d i n t e r a c t i o n s 175 P r o t e i n m i c e l l e s 101 P r o t e i n p r o c e s s i n g and t r a n s p o r t

259 P r o t e i n s y n t h e s i s 377 P r o t e o l y t i c enzymes 471 P u r p l e membrane 183

R a b i e s v i r u s 743 R a b b i t 679 R a b b i t B and T l y m p h o c y t e s 687 R a d i o c h e m i c a l a n a l y s i s 187 R a d i o i o d i n a t i o n 179 Radioimmunoassay 769, 873 R a d i o l a b e l l i n g 237 R a d i o s e q u e n c e o f p r o t e i n s 97 R a t 501, 615, 705 Ra t c e r e b r a l c o r t e x 535 Ra t serum g l y c o p r o t e i n 209 R a t i o b e t a / b e t a r e c e p t o r s 167 RCAI l e c t i n b i n d i n g 341 R e c e p t i v i t y 563 R e c e p t o r b i n d i n g 487 R e c e p t o r f o r heme 467 R e c e p t o r f o r p o l y a n i o n s 439 R e c e p t o r i n h i b i t i o n 471 R e c e p t o r i n t r i n s i c f a c t o r 385 R e c e p t o r - m e d i a t e d e n d o c y t o s i s 3 89,

427, 447, 463 R e c e p t o r s 477, 513, 595, 599 R e c e p t o r s f o r r e g u l a t o r y p e p t i d e s

595

SUBJECT INDEX 983

R e c y c l i n g 447 R e j e c t i o n 691 R e l a t i o n s t r u c t u r e f u n c t i o n 377 R e s e a l e d g h o s t 283 R e t r o v i r a l m a r k e r s 559 RIA 637 R i c i n 927 RNA o n c o g e n i c v i r u s e s 303

SA a n t i g e n 365 S a n d w i c h immunoassay f o r i n t e r f e r o n

763 S a n d w i c h t e s t 865 S c h i s t o s o m a mansoni 179 S c h i s t o s o m u l a 179 S c r e e n i n g p r o c e d u r e s 631, 637 S e c r e t i n 509, 513 S e c r e t o r y component 419 S e c r e t o r y v e s i c l e s 497 S e l e c t i o n 627 S e i f a s s o c i a t i o n 117 Semen 195 S e m i n a l p l a s m a 195 S e p a r a t i o n t e c h n i q u e s 175 S e q u e n t i a l i m m u n o d e p l e t i o n 893 Serum b a c t e r i c i d a l e f f e c t 317 Serum DBP 373 Serum p r o t e i n b i n d i n g 961 Sex S t e r o i d s 567 Sheep e r y t h r o c y t e membranes 267 Sheep r e d c e l l s 671 S i a l i c a c i d 57, 75, 365 S i a l o g l y c o p r o t e i n 49 Smooth m u s c l e c o n t r o l 527 Sodium d o d e c y l S u l f a t e - p o l y - a c r y l a -

mide g r a d i e n t e l e c t r o p h o r e s i s 183 Sodium D o d e c y l s u l p h a t e P o l y a c r y l a ­

mide s l a b g e l 911 S o f t a g a r c l o n i n g 627, 645 S o l u b l e a c t i n f r a c t i o n 373 S o l u b l e immune c o m p l e x e s 431 S p e c i f i c a c t i n b i n d i n g 373 S p e c i f i c b i n d i n g d e m o n s t r a t i o n 543 S p e c i f i c b i n d i n g S i t e s 527, 535,

539 S p e c i f i c d r u g c a r r i e r s 389 S p e c t r i n 121, 129 S p e c t r i n - p h o s p h o l i p i d i n t e r a c t i o n

127 S p e c t r o s c o p y 467 S p e r m a t o z o a 195 S p o n t a n e o u s l y h y p e r t e n s i v e (SHR)

r a t s 531 Ss a n t i g e n s 57 S t e r e o s p e c i f i c i t y 263 S t e r o i d hormones 823 S t r e p t o c o c c a l g r o u p A 797

S t r e p t o c o c c a l g r o u p i n g 747 S t r u c t u r e - a c t i v i t y - r e l a t i o n s h i p s 393 S t r u c t u r e - a c t i v i t y s t u d i e s 483 S u b a c u t e s c l e r o s i n g p a n e n c e p h a l i t i s

965 S u b s e t s 713 S u b s t a n c e P 477 S u b t y p e s 743 S u b u n i t e i n t e r a c t i o n 755 S u b u n i t V a c c i n e 101 S u p p r e s s i o n 671 S u p p r e s s o r c e l l s 683 S u p p r e s s o r f a c t o r 409 S u p p r e s s o r / c y t o t o x i c c e l l s 653 S u p p r e s s o r T c e l l s 663 S u r f a c e a n t i g e n s 179, 247 S u r f a c e g l y c o p e p t i d e s 75 S u r f a c e g l y c o p r o t e i n s 63 SV 40 159 S y n a p t i c p l a s m a membrane 151 Sy n a p t o s o m e s 535 S y n c y t i u m f o r m i n g u n i t s 559 S y n e r g i s m 751 S y n t h e t i c n e u r o t e n s i n a n a l o g u e s 483

T 3 817 T a r g e t i n g 921 T c e l l d i f f e r e n t i a t i o n 675 T c e l l h y b r i d 671 T c e l l s u b s e t s 667 T c e l l s 6 83, 705, 729 T c e l l s c y t o t o x i c 663 T e r n a r y c o m p l e x a t i o n 283 T e t a n u s t o x i n 247 T h e o r y o f immune compl e x f o r m a t i o n

649 T h y m o c y t e s 679, 7 0 1 , 705 T h y r o g l o b u l i n 861 T h y r o i d 579 T i s s u e d i s t r i b u t i o n 777 T l y m p h o c y t e s 653, 659, 701 T - l y m p h o c y t e s u b s e t s 691 T o r p e d o e l e c t r i c o r g a n 155, 793 TPA 143 T r a n s c e l l u l a r t r a n s p o r t o f IgA 423 T r a n s f e r e l e c t r o p h o r e s i s 965 T r a n s f e r r i n 459 T r a n s f e r r i n i r o n 447 T r a n s f e r r i n r e c e p t o r 227, 451,

455 T r a n s f o r m a t i o n 299, 451 T r a n s f o r m a t i o n a n t i g e n 739 T r a n s f o r m a t i o n e n h a n c i n g f a c t o r s

303 T r a n s f o r m e d l y m p h o c y t e s 739 Transmembrane i n t e r a c t i o n 287 Transmembrane l i n k a g e s 63

984 SUBJECT INDEX

T r i a z o l o p y r i d a z i n e 487 T r i t i u m 961 T r o p h o b l a s t 89 T r y p t i c p e p t i d e mapping 359 TSH r e c e p t o r 571, 575 Tumor a n t i g e n 899, 903 Tumor a s s o c i a t e d a n t i g e n s 93,

877, 889 Tumor c e l l s 159 Tumor s e r o l o g y 877 T u n i c a m y c i n 67 T w o d i m e n s i o n a l i m r n u n o e l e c t r o p h o r e -

s i s 961

U l t r a t h i n f r o z e n s e c t i o n s 773 U n i v a l e n t a n t i b o d i e s 313 U t e r u s 369

V a s o p r e s s i n r e c e p t o r s 517 V e s i c l e r e l e a s e 109 V i n d e s i n e 921 VIP 509, 513, 595 VIP r e c e p t o r s 535 V i r o s o m e s 101 V i r u s P o l y p e p t i d e s 965 V i s u a l i z a t i o n o f hormone b i n d i n g

555 V i t a m i n B 1 ^ r e c e p t o r 385 von W i l l e b r a n d f a c t o r 79

Wheat germ l e c t i n 89 Winn t e s t 159

Z u c k e r obese ( f a / f a ) r a t s 531 Z w i t t e r i o n i c d e t e r g e n t 33