Enzyme Kinetics

ENZYME KINETICS

This textbook for advanced courses in enzyme chemistry and enzyme kinetics covers the field of steady state enzyme kinetics from the basic principles inherent in the Henri equation to the expressions which describe the control of multi-enzyme pathways. Steady state kinetic equations are derived with the use of the connection matrix method, and an algorithm which can beimplemented easily for computer-based derivation of the equations is developed. Throughout the book emplasis is placed on a proper interpretation of the kinetic behavior of multi-reactant enzymic reactions. The principles of analytical geometry and basic calculus are employed to analyze non-hyperbolic substrate saturation curves in terms of the mechanisms, including subunit interactions, which cause thedeviation from hyperbolic curves. Sensitivity theory is employed to obtain expressions forfluxand concentration control coefficients for multi-enzyme pathways. The algorithm developed can be incorporated into a computer-based program for derivation. Emphasis is placed on the interpretation of control coefficients in terms of metabolic control. Problems are included at the end of each chapter andtheir solutions are found at the end of the book.

ENZYME KINETICS
From diastase to multi-enzyme systems
ARTHUR R. SCHULZ
Indiana University School of Medicine

CAMBRIDGE
UNIVERSITY PRESS

Published by the Press Syndicate of the University of Cambridge The Pitt Building, Trumpington Street, Cambridge CB2 1RP 40 West 20th Street, New York, NY 10011-4211, USA 10 Stamford Road, Oakleigh,Melbourne 3166, Australia © Cambridge University Press 1994 First published 1994
Library of Congress Cataloging-in-Publication Data

Schulz, Arthur R. Enzyme kinetics from diastase to multi-enzyme systems / Arthur R. Schulz p. cm. ISBN 0-521-44500-0. - ISBN 0-521-44950-2 (pbk.) 1. Enzyme kinetics. I. Title. QP601.3.S38 1994 94-28309 574.19'25-dc20 CIP
A catalog record for this book is availablefrom the British Library

ISBN 0-521-44500-0 Hardback ISBN 0-521-44950-2 Paperback Transferred to digital printing 2003

Contents

Preface

page ix 3 3 7 9 12 13 14 15 18 20 22 22 23 25 27 28 28 30 30

Part One: Basic steady state enzyme kinetics 1 Derivation of a rate equation 1.1 The role of 'diastase' in the early development of a theory 1.2 The basic assumptions on which derivationof an equation is based 1.3 The Briggs-Haldane steady state treatment of enzyme kinetic behavior 1.4 Estimation of steady state parameters 1.5 Problems for chapter 1 Appendix: A brief look at statistical analysis l.A.l Definition of a few statistical terms 1.A.2 Linear regression 1.A.3 Non-linear regression in enzyme kinetic analysis References for chapter 1 2 A closer look at the basicassumptions 2.1 Why must the substrate concentration greatly exceed that of the enzyme? 2.2 What if the substrate concentration does not greatly exceed that of the enzyme? 2.3 Examination of the entire time-course of an enzymic reaction 2.4 A precise definition of steady-state velocity 2.5 Problems for chapter 2 References for chapter 2 3 Enzyme inhibition 3.1 A general model of enzyme inhibition

vi3.2

Contents 34 35 37 38 41 42 43 44 44 50 52 53 55 56 59 59 61 69 75 76 77 77 82 87 91 91 92 92 94 100 102 103

Quantitative estimates of steady state parameters in enzyme inhibition 3.3 Competitive inhibition: A limiting case of inhibition 3.4 Uncompetitive inhibition: A different limiting case 3.5 Substrate inhibition 3.6 Problems for chapter 3 Appendix: A brief discussion of polyonomialregression References for chapter 3 4 Reversible enzyme-catalyzed reactions 4.1 Derivation of a rate equation by matrix inversion 4.2 Reformulation of the complete rate equation 4.3 The effect of product inhibition 4.4 Use of the King-Altman method to derive the rate equation 4.5 Problems for chapter 4 References for chapter 4 Part Two: Enzyme reaction sequence 5 Multi-reactant enzymic reactions...