Enzymes of the Cholinesterase Family

Enzymes of the Cholinesterase Family

Edited by

Daniel M. Quinn University of Iowa Iowa City, Iowa

A. S. Balasubramanian Christian Medical College and Hospital Vellore, Tamil Nadu, India

Bhupendra P. Doctor Walter Reed Army Institute of Research Washington, D.C.

and

Palmer Taylor University of California at San Diego La Jolla, California

Springer Science+Business Media, LLC

Library of Congress Cataloglng-ln-PublIcatIon Data

Enzymes of the C h o l i n e s t e r a s e family / ed i t e d by Daniel M. Quinn ... [e t a l . ] .

p. cm. "Proceedings of the F i f t h I n t e r n a t i o n a l Meeting on

C h o i i n e s t e r a s e s , held September 24-28, 1994, in Madras, I n d i a " — T . p . verso.

Includes b i b l i o g r a p h i c a l r e f e r e n c e s and index. ISBN 978-1-4899-1053-0 1. C h o i i n e s t e r a s e s — C o n g r e s s e s . I. Quinn, Daniel M.

I I . I n t e r n a t i o n a l Meeting on C h o i i n e s t e r a s e s (5th : 1994 : Madras, Indi a ) [DNLM: 1. A c e t y l c h o l i n e s t e r a s e — m e t a b o l i s m — c o n g r e s s e s .

2. B u t y r y l c h o l I n e s t e r a s e — m e t a b o l i s m — c o n g r e s s e s . QU 136E611 1995] QP609.C4E56 1995 574. 19'253~dc20 DNLM/DLC for Library of Congress 95-39179

CIP

Front cover: A model of the complex between the snake venom toxin fasciculin (light color) and acetylcholinesterase (dark color), constructed by H.K.L. van den Born et ai [(1995) Protein Science 4, 703—715]. Photography by Z. Radie of the University of California at San Diego.

Proceedings of the Fifth International Meeting on Cholinesterases, held September 24-28, 1994, in Madras, India

ISBN 978-1-4899-1053-0 ISBN 978-1-4899-1051-6 (eBook) DOI 10.1007/978-1-4899-1051-6

© Springer Science+Business Media New York 1995 Originally published by Plenum Press, New York in 1995 Softcover reprint of the hardcover 1st edition 1995

All rights reserved

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No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise, without written permission from the Publisher

PREFACE

The Fifth International Meeting on Cholinesterases convened in Madras, India, in September of 1994. The long and rich history and culture of India provided an excellent setting for the meeting. More than 120 delegates from Asia, Australia, Europe and North America heard 54 oral presentations and viewed 54 posters on current research on enzymes of the cholinesterase family. The aim of this book is to compile the presentations of the Fifth International Meeting on Cholinesterases into a volume that describes recent investigations on the structure and catalytic function of acetylcholinesterase (AChE), butyrylcholinesterase (BuChE) and related enzymes, as well as studies on the molecular and cellular biology of these enzymes and the genes which encode them.

Cholinesterases enjoy a long and storied history in diverse areas. In basic biochemical research, AChE is one of the best studied, though yet enigmatic, of enzymes. The efficient catalytic function of this enzyme presents the biochemist with a fundamental challenge in understanding the relationship between structure and function. AChE and BuChE belong to a family of proteins, the alB hydrolase fold family, whose constituents evolutionarily diverged from a common ancestor. Proteins in this family have a wide range of physiological functions. In commerce, AChE is a prime target for agricultural insect control, and for the development of therapeutic agents for Alzheimer's disease. On the national security front, AChE is the target of chemical warfare agents, "nerve gases," which pose a threat on the battlefield of nations at war and in the city centers of nations at peace. Recent events, chronicled in the international news media, provide a stark reminder of this fact.

The broad reach of cholinesterase function and research affects the biological scientist and nonscientist alike. From biotechnology to the military, one is hard pressed to find a family of enzymes whose effect on the human condition is more pervasive. Various presentations at the meeting described structure-activity relationships for the interaction of cholinesterases with toxic organophosphorus agents. Novel efforts at antidotal therapy were described that aim to use cholinesterases in the presence of nucleophilic oximes to detoxify organophosphorus inhibitors. Strategies for effective inhibition of acetylcholinesterase in the central nervous system, as an approach to the treatment of the cognitive dis functions associated with Alzheimer's disease, were described, as were the structure-activity relation­ships of new anti-Alzheimer's agents.

The cell and molecular biology of cholinesterases is advancing rapidly. The relation­ships between AChE gene structure and the various molecular forms of the enzyme were described at the meeting, as were the mechanisms of gene expression, cellular biosynthesis, and assembly of the cholinesterases. Various noncatalytic roles for cholinesterases were described, which include involvement in nervous system development and in neurotrans­mission involving dopamine-containing nerve terminals.

v

vi Preface

Reports of structure-function relationships not only described the fine detail that is emerging on the structural basis of the rapid catalytic mechanisms effected by choli­nesterases, but also provided the bases for several controversies. X-ray crystallography of catalytically relevant ligand complexes with AChE provided visual verification of the roles, suggested by site-directed mutagenesis and other studies, for loci in the active site that are responsible for acyl group specificity, quaternary ammonium recognition, oxyanion recog­nition and acid-base catalysis. The crystal structure of AChE provided the template for theoretical evaluation ofthe role of the electrical field of the enzyme in ligand binding. Good spirited controversies arose on whether the electrical field of the enzyme plays a significant role in the rapidity of AChE catalysis, and whether an anthropomorphic "back door" mechanism is required for release of cationic products.

This Preface outlines just a fraction of the wide range of topics discussed at the Fifth International Meeting on Cholinesterases in Madras, India. The chapters that follow provide a clear view of the breadth and vigor of research on enzymes of the cholinesterase family. The meeting in Madras was a once in a lifetime opportunity for an international delegation to discuss and debate such research in a congenial atmosphere. Our Indian hosts are to be commended for the graciousness and organizational effort that were key to the success of the meeting.

Daniel M. Quinn A. s. Balasubramanian Bhupendra P. Doctor Palmer Taylor

CONTENTS

Part I. GENE STRUCTURE AND EXPRESSION OF CHOLINESTERASES

A. Presentations

Antisense Oligonucleotides Suppressing Expression of Cholinesterase Genes Modulate Hematopoiesis in Vivo and ex Vivo ....................... .

Hermona Soreq, Efrat Lev-Lehman, Deborah Patinkin, Mirta Grifman, Gal Ehrlich, Dalia Ginzberg, Fritz Eckstein, and Haim Zakut

Properties of Class A Acetylcholinesterase, the Enzyme Encoded by ACE-I in Caenorhabditis elegans . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

Martine Arpagaus, Nathalie Schirru, Emmanuel Culetto, Vincenzo Talesa, Xavier Cousin, Arnaud Chatonnet, Yann Fedon, Jean-Baptiste Berge, Didier Fournier, and Jean-Pierre Toutant

Ligand Specificity and Gene Expression in the Cholinergic Synapse: A Comparison between Acetylcholinesterase and the Nicotinic Acetylcholine Receptor.. 15

Palmer Taylor, Zoran Radic, Hans-JUrgen Kreienkamp, Zhigang Luo, Natilie A. Pickering, and Shelley Camp

Butyrylcholinesterase Transcription Start Site and Promoter ..................... 23 Omar Jbilo, Jean-Pierre Toutant, Arnaud Chatonnet, and Oksana Lockridge

The C-Terminal Alternative Regions of Acetylcholinesterase. . . . . . . . . . . . . . . . . . . .. 29 Jean Massoulie, Alain Anselmet, Suzanne Bon, Fran<,:oise Coussen,

Eric Krejci, and Claire Legay

Developmental Expression of Acetylcholinesterase in Skeletal Muscle. . . . . . . . . . . .. 37 Zoran Grubic and Armand F. Miranda

vii

viii Contents

B. Posters

Alternative Exon 6 Directs Synaptic Localization of Recombinant Human Acetylcholinesterase in Neuromuscular Junctions of Xenopus laevis Embryos ..................................................... 45

Meira Sternfeld, Shlomo Seidman, Revital Ben Aziz-Aloya, Michael Shapira, Rina Timberg, Daniela Kaufer, and Hermona Soreq

Developmental Regulation of Acetylcholinesterase mRNA in the Mouse Diaphragm: Alternative Splicing and Focalization ............................. 47

Claire Legay, Monique Huchet, Jean Massoulit\ and Jean-Pierre Changeux

Acetylcholinesterase and Butyrylcholinesterase Expression in Adult Rabbit Tissues and during Development . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 49

Omar Jbilo, Yann L'Hermite, Vincenzo Talesa, Jean-Pierre Toutant, and Arnaud Chatonnet

Mutations in the Catalytic Subunit of Acetylcholinesterase Do Not Appear Responsible for Congenital Myasthenic Syndrome Associated with End-Plate Acetylcholinesterase Deficiency ......................... 51

S. Camp, A. G. Engel, D. K. Getman, S. Bon, J. Massoulie, and P. Taylor

Regulation of Acetylcholinesterase (AChE) mRNA by Ryanodine-Sensitive and L-Type Calcium Channels during Myogenesis in Vitro and Muscle Development in Vivo . .......................................... 53

Zhigang Luo, Martine Pincon-Raymond, and Palmer Taylor

Regulation of AChE Gene Expression in Neuronally Induced Mouse P19 Cells. . . . .. 55 B. A. Coleman and P. Taylor

Regulation of Human Acetylcholinesterase Gene Expression. . . . . . . . . . . . . . . . . . . .. 56 D. Getman, K. Inoue, and P. Taylor

Promoter Elements of the Mouse Acetylcholinesterase Gene: Regulation during Muscle Differentiation ......................................... 57

Annick Mutero and Palmer Taylor

Part II. POLYMORPHISM AND STRUCTURE OF CHOLINESTERASES

A. Presentations

Structures of Complexes of Acetylcholinesterase with Covalently and Non-Covalently Bound Inhibitors ................................ 59

J. L. Sussman, M. Harel, M. Raves, D. M. Quinn, H. K. Nair, and I. Silman

Electrostatic Properties of Human Acetylcholinesterase. . . . . . . . . . . . . . . . . . . . . . . .. 67 Daniel R. Ripoll, Carlos H. Faerman, Richard Gillilan, Israel Silman, and

Joel L. Sussman

Contents ix

Substrate Binding Site and the Role of the FLAP Loop in Candida rugosa Lipase, a Close Relative of Acetylcholinesterase. . . . . . . . . . . . . . . . . . . . . . . . . . . .. 71

Miroslaw Cygler, Pawel Grochulski, and Joseph D. Schrag

Studies on Partially Unfolded States of Torpedo californica Acetylcholinesterase 77 Israel Silman, David I. Kreimer, Irina Shin, Elena A. Dolginova, Ester Roth,

Daniella Goldfarb, Reuven Szosenfogel, Mia Raves, Joel L. Sussman, Nina Borochov, and Lev Weiner

FTIR-Spectroscopic Investigations of the Structure and Temperature Stability of the Acetylcholinesterase from Torpedo californica .. . . . . . . . . . . . . . . . . . . .. 83

Ferdinand Hucho, Dieter Naumann, and Ute Gome-Tschelnokow

Residues in the C-Terminus of Torpedo californica Acetylcholinesterase Important for Modification into a Glycophospholipid Anchored Form . . . . . . . . . . .. 89

Goran Bucht, Lena Lindgren, and Karin Hjalmarsson

Computer Modeling of Acetylcholinesterase and Acetylcholinesterase-Ligand Complexes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 97

S. T. Wlodek, J. Antosiewicz, M. K. Gilson, 1. A. McCammon, T. W. Clark, and L.R. Scott

Structural Analysis of the Asparagine-Linked Oligosaccharides ofCholinesterases: N-Linked Carbohydrates of Cholinesterases ........................ 105

Ashima Saxena and B. P. Doctor

Pressure Effects on Structure and Activity of Cholinesterase ..................... 113 P. Masson and C. Clery

B. Posters

Hydrophobicity on Esterase Activity of Human Serum Cholinesterase ............. 123 L. Jaganathan, K. Padmalatha, G. Revathi, and R. Boopathy

Alterations in the Topography of Acetylcholinesterase Active Site Gorge after Binding of Peripheral Anionic Site Ligands ......................... 125

Anton Stalc, Zoran Grubic, Marjeta Sentjurc, Slavko Pecar , Mary K. Gentry, and Bhupendra P. Doctor

Acetylcholinesterase from Octopus vulgaris (Cephalopoda): Evidence for a Specific High Salt-Soluble and Heparin-Soluble Fraction of Globular Forms ..... 127

Vincenzo Talesa, Marta Grauso, Elvio Giovannini, Gabriella Rosi, and Jean-Pierre Toutant

Molecular Polymorphism of Acetylcholinesterase in Hirudo medicinalis ........... 128 Vincenzo Talesa, Marta Grauso, Elvio Giovannini, Gabriella Rosi, and

Jean-Pierre Toutant

Subunit Association and Stabilization of Butyrylcholinesterase (BChE) ............ 129 R. M. Blong, P. Masson, and O. Lockridge

x Contents

Denaturation of Recombinant Human Acetylcholinesterase ...................... 131 M. Lebleu, C. Clery, P. Masson, S. Reuveny, D. Marcus, B. Velan, and

A. Shafferman

Part III. MECHANISM OF CATALYSIS OF CHOLINESTERASES

A. Presentations

Amino Acid Residues that Control Mono- and Bisquatemary Oxime-Induced Reactivation of O-Ethyl Methylphosphonylated Cholinesterases ........ 133

Y. Ashani, Z. Radic, I. Tsigelny, D. C. Vellom, N. A Pickering, D. M. Quinn, B. P. Doctor, and P. Taylor

Modulation of Catalysis and Inhibition of Fetal Bovine Serum Acetylcholinesterase by Monoclonal Antibodies ...................................... 141

B. P. Doctor, Mary K. Gentry, Ashima Saxena, and Yacov Ashani

Insect Acetylcholinesterase and Resistance to Insecticides ....................... 149 Didier Fournier, Marie Maturano, Laurent Gagnoux, Philippe Ziliani,

Cyril Pertuy, Madeleine Pralavorio, Jean-Marc Bride, Leila Elmarbouh, Alain Klaebe, and Patrick Masson

Phosphonate Ester Active Site Probes of Acetylcholinesterase, Trypsin and Chymotrypsin ................................................ 155

Akos Bencsura, Istvan Enyedy, Carol Viragh, Rinat Akhmetshin, and Ildiko M. Kovach

Irreversible Site-Directed Labeling Studies on Cholinesterases ................... 163 L. Ehret-Sabatier, I. Schalk, C. Loeb, F. Nachon, and M. Goeldner

Mutation of Human Butyrylcholinesterase Glycine 117 to Histidine Preserves Activity but Confers Resistance to Organophosphorus Inhibitors ....... 169

C. A Broomfield, C. B. Millard, o. Lockridge, and T. L. Caviston

Reaction of Acetylcholinesterase with Organophosphonates: Molecular Fate at the Rim of a Gorge ............................................... 177

Harvey Alan Berman

Amino Acid Residues in Acetylcholinesterase which Influence Fasciculin Inhibition .. 183 Zoran Radic, Daniel M. Quinn, Daniel C. Vellom, Shelley Camp, and

Palmer Taylor

Molecular Aspects of Catalysis and of Allosteric Regulation of Aceytlcholinesterases 189 A. Shaff erman, AOrdentlich, D. Barak, C. Kronman, N. Ariel, M. Leitner,

Y. Segall, A. Bromberg, S. Reuveny, D. Marcus, T. Bino, A. Lazar, S. Cohen, and B.Velan

Contents xi

Structural Determinants ofFasciculin Specificity for Acetylcholinesterase .......... 197 Pascale Marchot, Shelley Camp, Zoran Radic, Pierre E. Bougis, and

Palmer Taylor

The Function of Electrostatics in Acetylcholinesterase Catalysis .................. 203 Daniel M. Quinn, Javier Seravalli, Haridasan K. Nair, Rohit Medhekar,

Basel Husseini, Zoran Radic, Daniel C. Vellom, Natilie Pickering, and Palmer Taylor

Fasciculin 2 Binds to the Peripheral Site on Acetylcholinesterase and Inhibits Substrate Hydrolysis by Slowing a Step Involving Proton Transfer during Enzyme Acylation ....................................... 209

Jean Eastman, Erica J. Wilson, Carlos Cervenansky, and Terrone L. Rosenberry

B. Posters

New Photolabile Inhibitors ofCholinesterases Designed for Rapid Photochemical Release of Choline ............................................. 219

L. Peng and M. Goeldner

Amino Acids Determining Specificity to OP-Agents and Facilitating the "Aging" Process in Human Acetylcholinesterase ............................ 221

A. Ordentlich, C. Kronman, D. Stein, N. Ariel, S. Reuveny, D. Marcus, y. Segall, D.Barak, B. Velan, and A.Shafferman

Electrostatic Attraction by Surface Charge does not Contribute to the Catalytic Efficiency of Acetylcholinesterase ................................ 223

D. Barak, A. Ordentlich, C. Kronman, R. Ber, T. Bino, N.Ariel, R. Osman, B. Velan, and A. Shafferman

Catalytic Properties of Human Serum Cholinesterase Phenotypes in their Reaction with Substrates and Inhibitors .................................... 225

V. Simeon-Rudolf, M. Skrinjaric-Spoljar, and E. Reiner

Organophosphate Specificity of Acyl Pocket Cholinesterase Mutants ............... 227 Natilie A. Pickering, Palmer Taylor, and Harvey Berman

London Dispersion Interactions in Molecular Recognition by Acetylcholinesterase ... 228 Daniel M. Quinn, Haridasan K. Nair, Javier Seravalli, Keun Lee,

Tomira Arbuckle, Zoran Radic, Daniel C. Vellom, Natilie Pickering, and Palmer Taylor

Peripheral Anionic Site of Wild-Type and Mutant Human Butyrylcholinesterase ..... 230 P. Masson, M.T. Froment, C. Bartels, and o. Lockridge

xii Contents

Part IV. CELLULAR BIOLOGY OF CHOLINESTERASES

A. Presentations

Dual Control of Acetylcholinesterase Content of Mature Innervated Fast Muscles .... 233 Victor Gisiger

Restricted Regulation of AChE Transcription, Translation, and Localization: Individual Nuclei Respond to Locally Generated Signals in Multinucleated Skeletal Muscle Fibers ............................. 239

Richard L. Rotundo, Susana G. Rossi, Rosely O. Godinho, Ana E. Vazquez, and Bhavya Trivedi

Mechanism and Implications of Selective Neural Lesions by Acetylcholinesterase Antibodies in the Central and Peripheral Nervous Systems: AChE Autoimmunity ................................................ 247

S. Brimijoin, P. Hammond,and Z. Rakonczay

Neural Regulation of Acetylcholinesterase Expression in Slow and Fast Muscles of the Rat ...................................................... 253

J. Sketelj and B. Cresnar

Neuromuscular Factors Influencing Acetylcholinesterase Gene Expression in Skeletal Muscle Fibers ......................................... 261

B. J. Jasmin, C. Boudreau-Lariviere, R. Chan, D. A. Hubatsch, and H. Sveistrup

Post-Translation Processing of Acetylcholinesterase: Cellular Control of Biogenesis and Secretion ................................................. 269

Baruch Velan, Chanoch Kronman, Arie Ordentlich, Yehuda Flashner, Raphael Ber, Sara Cohen, and Avigdor Shafferman

Acetylcholinesterase at Neuromuscular Junctions: Density, Catalytic Turnover, and Molecular Forms with Modeling and Physiological Implications ........ 277

L. Anglister, J. R. Stiles, B. Haesaert, J. Eichler, and M. M. Salpeter

B. Posters

Effect of Acute Irreversible Inhibition of Acetylcholinesterase on Nerve Stimulation-Evoked Contractile Properties of Rat Medial Gastrocnemius 287

R. Panenic, P. Gardiner, and V. Gisiger

Developmental Regulations of Acetylcholinesterase at the Molecular Level in the Central Nervous System of the Quail .............................. 288

Alain Anselmet, Jean-Marc Chatel, and Jean Massoulie

Contents xiii

Reversal of Glycerol Ether-Stimulated Acetylcholinesterase Activity by c-fos Antisense Oligonucleotide in Primary Neuronal Cultures .............. 289

J. R. Dave, E. Fasnacht, F. C. Tortella, J. M. Best, B. P. Doctor, and H. S. Ved

Acetylcholinesterase Activity in Plants ...................................... 291 S. Madhavan, Gautam Sarath, and Patricia L. Herman

Signal-Mediated Cellular Retention and Subunit Assembly of Human Acetylcholinesterase ........................................... 293

C. Kronman, Y. Flashner, A. Shaff erman, and B. Velan

The Grain Aphid Cholinesterases ........................................... 295 B. Leszczynski, T. Bakowski, A.F.G. Dixon, and H. Matok

Part V. STRUCTURE-FUNCTION RELATIONSHIPS OF ANTICHOLINESTERASE AGENTS

A. Presentations

Review of Nerve Agent Inhibitors and Reactivators of Acetylcholinesterase ......... 297 David H. Moore, Charles B. Clifford, Isabelle T. Crawford, Greg M. Cole,

and Jack M. Baggett

Organophosphate-Sensitive Cholinergic Receptors: Possible Role in Modulation of Anticholinesterase-Induced Toxicity .............................. 305

C. N. Pope, J. Chaudhuri, and T. K. Chakraborti

Effect of Physic all Chemical Stressors on the Cholinergic System in Rat ............ 313 Satu M. Somani

Evaluation of the Direct Actions ofHI-6 in Reversing Soman-Induced Tetanic Fade .. 321 Michael Adler, Donald M. Maxwell, Richard E. Sweeney, and

Sharad S. Deshpande

Clinical Blood Cholinesterase Measurements for Monitoring Pesticide Exposures .... 329 B. W. Wilson, S. Padilla, J. R. Sanborn, J. D. Henderson, and J. E. Billitti

Repeated Dosing with Chlorpyrifos Increases Acetylcholinesterase Immunoreactivity in Rat Brain ................................... 337

S. Padilla, S. Chiappa, C. Koenigsberger, V. Moser, and S. Brimijoin

Bisquaternary Oximes as Antidotes against Tabun and Soman Poisoning: Antidotal Efficacy in Relation to Cholinesterase Reactivation .................. 345

G. Amitai, I. Rabinovitz, G. Zomber, R. Chen, G. Cohen, R. Adani, and L. Raveh

xiv Contents

Comparison of Acetylcholinesterase, Pyrldostigmine, and HI-6 as Antidotes against Organophosphorus Compounds .................................. 353

Donald M. Maxwell, Karen M. Brecht, Ashima Saxena, Palmer Taylor, and Bhupendra P. Doctor

Inhalation Toxicokinetics ofC(±)P(±)-Soman and (±)-Sarin in the Guinea Pig ....... 361 H. P. Benschop, L. P. A. de Jong, and 1. P.Langenberg

Inhibition ofFish Brain Acetylcholinesterase by Cadmium and Mercury: Interaction with Selenium ................................................ 369

S. Sen, S. Mondal, 1. Adhikari, D. Sarkar, S. Bose, B. Mukhopadhyay, and Shelley Bhattacharya

Pharmacological Evaluation of Antidotes against Organophosphorus Intoxication .... 375 S. N. Dube, R. Bhattacharya, K. Husain and A. K. Sikder

Medical Protection against Organophosphorus Toxicity ......................... 381 S. Das Gupta

B. Posters

Identification of a 155 kDa Fraction That Possesses Neuropathy Target Esterase Activity ..................................................... 387

C. E. Mackay, B. D. Hammock, and B. W. Wilson

Interleukin 6 Modulates AChE Activity in CNS Neurons ........................ 389 D. Claren90n, E. Multon, M. Galonnier, M. Estrade, C. Fournier,

J. Mathieu, J. C. Mestries, G. Testylier, P. Gourmelon, and A. Fatome

HI-6 is Incapable of Reactivating Tabun-Phosphonylated Human Acetylcholinesterase ........................................... 391

Chunyuan Luo, Jinsheng Yang, and Manji Sun

Pattern ofInhibition of Acetylcholinesterases in Different Regions of the Brain by Two Organophosphorus Homologues, in Relation to Their Differential Neurotoxicity ................................................. 392

P. Santhoshkumar, Subramanya Karanth, and T. Shivanandappa

Organophosphate-Sensitive Carboxyl esterase Isozyme(S) in the Rat Liver .......... 394 Subramanya Karanth and T. Shivanandappa

Fenthion Treatment Produces Tissue-, Dose-, and Time-Dependent Decreases in Muscarinic Second Messenger Response in the Adult Rat CNS ......... 396

P. Tandon, C. N. Pope, S. Barone, Jr., W. Boyes, H. A. Tilson, and S. Padilla

Protection of Guinea Pigs against Soman Inhalation by Pretreatment Alone with Human Butyrylcholinesterase .................................... 398

N. Allon, L. Raveh, E. Gilat, J. Grunwald, E. Manistersky, E. Cohen, and Y. Ashani

Contents xv

Human Butyrylcholinesterase as Prophylaxis Treatment against Soman: Behavioral Test in Rhesus Monkeys ........................................ 400

E. Grauer, L. Raveh, J. Kapon, J. Grunwald, E. Cohen, and Y. Ashani

Efficacy of Prophylaxis with Human Butyrylcholinesterase against Soman and VX Poisoning: A Comparative Analysis ............................... 402

L. Raveh, J. Grunwald, E. Cohen, and Y. Ashani

Prevention of Brain Damage and Behavioral Performance Changes following an IV· Injection of Soman and VX in Rats Pretreated with Human Butyrylcholinesterase .......................................... 404

T. Kadar, L. Raveh, R. Brandeis, J. Grunwald, E. Cohen, and Y. Ashani

Inactivation of the Catalytic Activities in Human Serum Butyrylcholinesterase by Metal Chelators ............................................... 406

C. D. Bhanumathy and A. S. Balasubramanian

Protective Efficacy of Physostigmine and Pyridostigmine at Various Pretreatment Times against Inhaled Sarin in Rats ............................... 408

R. Vijayaraghavan, K. Husain, P. Kumar, K. S. Pandey, and S. Das Gupta

Induction of Carboxylesterase Isoenzymes and Altered Acetylcholinesterase during OP Resistance in Mosquito Culex quinquefasciatus .................. 410

N. Gopalan, S. Prakash, K. M. Rao, and B. K. Bhattacharya

Structure-Activity ofPyridinium Oximes as Antidotes to Organophosphorus Anticholinesterase Agents ....................................... 412

A. K. Sikder, S. N. Dube, and D. K. Jaiswal

Part VI. NONCHOLINERGIC FUNCTIONS OF CHOLINESTERASES

A. Presentations

A Non-Cholinergic Function for Acetylcholinesterase .......................... 415 S. A. Greenfield

Non-Cholinergic Function of Cholinesterases ................................. 423 K. M. Kutty, V. Prabhakaran, and A. R. Cooper

Glycosylated Inactive Forms of Chicken Butyrylcholinesterases and Their Possible Functions .................................................... 427

Paul G. Layer, Christoph Ebert, Sven Treskatis, Thomas Weikert, and Elmar Willbold

Possible Cholinergic and Non-Cholinergic Actions of Transiently Expressed Acetylcholinesterase in Thalamocortical Development Projections ...... 435

Richard T. Robertson, Ron S. Broide, Jen Yu, and Frances L. Leslie

xvi Contents

B. Posters

Evidence for a Putative Acetylcholinesterase Uptake Mechanism within the Substantia Nigra .............................................. 443

T. C. Budd, B. G. M. Dickie, D. Vaux, and S. A. Greenfield

Activation of Peritoneal Macrophages by Acetylcholinesterase Is Independent of Catalytic Activity .............................................. 445

A. Klegeris, T. C. Budd, and S. A. Greenfield

Functional Significance ofButyrylcholinesterase for Formation of Neural Networks as Tested by Chicken Retinospheroids ............................. 447

Elmar Willbold and Paul G. Layer

Neurotrophic Function of Circulating Acetylcholinesterase in Ap/ysia ............. 449 M. Srivatsan and B. Peretz

Part VII. PHARMACOLOGICAL UTILIZATION OF ANTICHOLINESTERASES

A. Presentations

Cholinesterases in Alzheimer's Disease ...................................... 451 M.-Marsel Mesulam

A Role for Acetylcholine and Acetylcholinesterase in the Regulation of Neurite Outgrowth: Implications for Alzheimer's Disease .................... 455

D. H. Small and G. Reed

Cholinesterase Inhibitors: From Preclinical Studies to Clinical Efficacy in Alzheimer Disease ...................................................... 463

E. Giacobini

B. Posters

Genetic Predisposition for Variable Response to Anticholinesterase Therapy Anticipated in Carriers of the Butyrylcholinesterase "Atypical" Mutation 471

Y. Loewenstein, M. Schwarz, D. Glick, B. Norgaard-Pedersen, H. Zakut, andH. Soreq

Horse Serum Butyrylcholinesterase Does Not Disrupt Passive Avoidance Learning or Spontaneous Motor Activity in Rats ............................ 473

Raymond F. Genovese, Averi R. Roberts, William E. Fantegrossi, Roberta Larrison, and Bhupendra P. Doctor

Acetylcholinesterase Inhibitors (AChE-I) as a Potential Use for Alzheimer's Disease (AD) Therapy: Differences in Mechanisms of Enzyme Inhibition ....... 475

Albert Enz

Contents xvii

Comparative Inhibition of Acetylcholinesterase by Tacrine, Physostigmine, and Huperzine in the Adult Rat Brain ................................. 477

H. S. Ved, 1. M. Best, J. R. Dave, and B. P. Doctor

Acetylcholinesterase Gene Sequence and Copy Number are Normal in Alzheimer's Disease Patients Treated with the Organophosphate Metrifonate ........ 479

C. F. Bartels, P. L. Moriearty, R. E. Becker, C. P. Mountjoy, and O. Lockridge

Part VIII. APPENDICES

Appendix I

Compilation of Evaluated Mutants of Cholinesterases .......................... 481 A. Shaff erman, C. Kronman, and A. Ordentlich

Appendix II

A Database of Sequences Related to AcetylcholinesteraselLipase/a:p Hydrolase Superfamily with Public Access on Internet ........... ' .............. 489

Xavier Cousin, Thierry Hotelier, Catherine Mazzoni, Martine Arpagaus, Jean-Pierre Toutant, and Arnaud Chatonnet

Appendix III

Amino Acid Alignment of Cholinesterases, Esterases, Lipases, and Related Proteins .. 493 Mary K. Gentry and B. P. Doctor

Appendix IV

Participants ............................................................ 507

Author Index ........................................................... 517

Subject Index ........................................................... 521