Now in its fourth edition, this textbook is one of the few titles worldwide to cover enzyme kinetics in its entire scope and the only one to include its implications for bioinformatics and systems biology. Multi-enzyme complexes and cooperativity are therefore treated in more detail than in any other textbook on the market.
The respected and well known author is one of the most experienced researchers into the topic and writes with outstanding style and didactic clarity. As with the previous editions, he presents here steady-state kinetics and fast reactions, supplementing each chapter with problems and solutions. For the first time, this edition features color illustrations and a companion website.
TOC:
1 Basic Principles of Chemical Kinetics
1.1 Abbreviations and symbols
1.2 Order of a reaction
1.3 Dimensions of rate constants
1.4 Reversible reactions
1.5 Determination of first-order rate constants
1.6 Catalysis
1.7 The influence of temperature and pressure on rate constants
Problems
2 Introduction to Enzyme Kinetics
2.1 The idea of an enzyme?substrate complex
2.2 The Michaelis?Menten equation
2.3 The steady state of an enzyme-catalysed reaction
2.5 Validity of the steady-state assumption
2.6 Graphs of the Michaelis?Menten equation
2.7 The reversible Michaelis?Menten mechanism
2.8 Product inhibition
2.9 Integration of enzyme rate equations
2.10 Artificial enzymes, RNA enzymes and catalytic antibodies
Problems
3 Practical Aspects of Kinetic Studies
3.1 Enzyme assays
3.2 Estimating the initial rate
3.3 Detecting enzyme inactivation
3.4 Experimental design
3.5 Treatment of ionic equilibria
Problems
4 Deriving Steady-State Rate Equations
4.1 Introduction
4.2 The principle of the King?Altman method
4.3 The method of King and Altman
4.4 The method of Wong and Hanes
4.5 Modifications to the King?Altman method
4.6 Reactions containing steps at equilibrium
4.7 Analysing mechanisms by inspection
4.3 The method of King and Altman
4.4 The method of Wong and Hanes
4.5 Modifications to the King?Altman method
4.6 Reactions containing steps at equilibrium
4.7 Analysing mechanisms by inspection
4.8 A simpler method for irreversible reactions
4.9 Derivation of rate equations by computer
Problems
5 Reversible Inhibition and Activation
5.1 Introduction
5.2 Linear inhibition
5.3 Plotting inhibition results
5.4 Multiple inhibitors
5.5 Relationship between inhibition constants and the concentration for 50% inhibition
5.6 Inhibition by a competing substrate
5.7 Enzyme activation
5.8 Design of inhibition experiments
5.9 Inhibitory effects of substrates
Problems
6 Tight-binding and Irreversible Inhibitors
6.1 Tight-binding inhibitors
6.2 Irreversible inhibitors
6.3 Substrate protection experiments
6.4 Mechanism-based inactivation
6.5 Chemical modification as a means of identifying essential groups
6.6 Inhibition as the basis of drug design
Problems
7 Reactions of More than One Substrate
7.1 Introduction
7.2 Classification of mechanisms
7.3 Rate equations
7.3.4 Haldane relationships
7.4 Initial-rate measurements in the absence of products
7.5 Substrate inhibition
7.6 Product inhibition
7.7 Design of experiments
7.8 Reactions with three or more substrates
Problems
8 Use of Isotopes for Studying Enzyme Mechanisms
8.1 Isotope exchange and isotope effects
8.2 Principles of isotope exchange
8.3 Isotope exchange at equilibrium
8.4 Isotope exchange in substituted-enzyme mechanisms
8.5 Non-equilibrium isotope exchange
8.6 Theory of kinetic isotope effects
8.7 Primary isotope effects in enzyme kinetics
8.8 Solvent isotope effects
Problems
9 Effect of pH on Enzyme Activity
9.1 Enzymes and pH
9.2 Acid?base properties of proteins
9.3 Ionization of a dibasic acid
9.4 Effect of pH on enzyme kinetic constants
9.5 Ionization of the substrate
9.6 More complicated pH effects
Problems
10 Temperature Effects on Enzyme Activity
10.1 Temperature denaturation
10.2 Temperature optimum
10.3 Application of the Arrhenius equation to enzymes
10.4 Entropy?enthalpy compensation
Problems
11 Regulation of Enzyme Activity
11.1 Function of cooperative and allosteric interactions
11.2 The development of models to explain cooperativity
11.3 Analysis of binding experiments
11.4 Induced fit
11.5 The symmetry model of Monod, Wyman and Changeux
11.6 Comparison between the principal models of cooperativity
11.7 The sequential model of Koshland, Nemethy and Filmer
11.8 Kinetic cooperativity
Problems
12 Kinetics of Multi-Enzyme Systems
12.1 Enzymes in their physiological context
12.2 Metabolic control analysis
12.3 Elasticities
12.4 Control coefficients
12.5 Properties of control coefficients
12.5.2 Implications for large changes
12.5.3 Constrained enzyme concentrations
12.6 Relationships between elasticities and control coefficients
12.7 Response coefficients: the partitioned response
12.8 Control and regulation
12.9 Mechanisms of regulation
12.10 Computer modelling of metabolic systems
12.11 Implications for biotechnology and drug discovery
Problems
13 Fast Reactions
13.1 Limitations of steady-state measurements
13.2 Product release before completion of the catalytic cycle
13.3 Experimental techniques
13.4 Transient-state kinetics
Problems
14 Estimation of Kinetic Constants
14.1 The effect of experimental error on kinetic analysis
14.2 Least-squares fit to the Michaelis?Menten equation
14.2.4 Estimating weights from replicate observations
14.3 Statistical aspects of the direct linear plot
14.4 Precision of estimated kinetic parameters
14.4.4 Km as the least precise Michaelis?Menten parameter
14.5 Generalizing the results to more than two parameters
14.6 Residual plots and their uses
Problems
APPENDIX: Standards for Reporting Enzymology Data
Solutions and Notes to Problems