Principles of Enzyme Kinetics

1 Basic Principles of Chemical Kinetics 1.1 Order of Reaction 1.2 Determination of the Order of a Reaction 1.3 Dimensions of Rate Constants 1.4 Reversible Reactions 1.5 Determination of First-Order Rate Constants 1.6 Influence of Temperature on Rate Constants 1.7 Transition-State Theory2 Introduction to Enzyme Kinetics 2.1 Early Studies 2.2 Work of Michaelis and Menten 2.3 Steady-State Treatment 2.4 Validity of the Steady-State Assumption 2.5 Graphical Representation of the Michaelis-Menten Equation 2.6 Reversible Michaelis-Menten Mechanism 2.7 Product Inhibition Appendix 2.1 Hyperbolic Nature of the Michaelis-Menten Equation3 How to Derive Steady-State Rate Equations 3.1 Introduction 3.2 Principle of the King-Altman Method 3.3 Method of King and Altman 3.4 Modifications to the King-Altman Method 3.5 Compression of Patterns 3.6 Reactions Containing Steps at Equilibrium 3.7 Analysing Mechanisms by Inspection 3.8 Rate Equations in Coefficient Form4 Inhibitors and Activators 4.1 Reversible and Irreversible Inhibitors 4.2 Competitive Inhibition 4.3 Mixed Inhibition 4.4 Uncompetitive Inhibition 4.5 Plotting Inhibition Results 4.6 Intuitive Approach to Linear Inhibition 4.7 Hyperbolic Inhibition and Activation 4.8 Non-Productive Binding 4.9 Substrate Inhibition 4.10 Inhibitors of High Affinity5 Reaction Pathways 5.1 Introduction 5.2 Survey of Two-Substrate, Two-Product Reaction Mechanisms 5.3 Nomenclature and Schematic Representation of Mechanisms 5.4 Rate Equations 5.5 Initial-Velocity Measurements in Absence of Products 5.6 Substrate Inhibition 5.7 Reverse Reaction 5.8 Product Inhibition 5.9 Isotope Exchange 5.10 Induced Transport6 Effects of pH and Temperature on Enzymes 6.1 pH and Enzyme Kinetics 6.2 Ionization of a Dibasic Acid 6.3 Effect of pH on Enzyme Kinetic Constants 6.4 pH Independence of Km 6.5 Ionization of Groups Remote from the Active Site 6.6 Change of Rate-Determining Step with pH 6.7 Temperature Dependence of Enzyme-Catalysed Reactions 6.8 Use of Temperature for Studying Enzyme Specificity7 Control of Enzyme Activity 7.1 Necessity for Metabolic Control 7.2 Binding of Oxygen to Haemoglobin 7.3 Hill Equation 7.4 Adair Equation 7.5 Pauling's Treatment 7.6 Induced Fit 7.7 Symmetry Model of Monod, Wyman and Changeux 7.8 Sequential Model of Koshland, Nemethy and Filmer 7.9 Half-of-the-Sites Reactivity 7.10 Other Equilibrium Models of Co-Operativity 7.11 Kinetic Models of Co-Operativity8 Analysis of Progress Curves 8.1 Integrated Rate Equations 8.2 Integrated Michaelis-Menten Equation 8.3 Competitive Product Inhibition 8.4 Inhibition by Several Products 8.5 Mixed Inhibition by Products 8.6 More Complex Cases 8.7 Some Pitfalls9 Fast Reactions 9.1 Limitations of Steady-State Measurements 9.2 Transient Phase of the Michaelis-Menten Mechanism 9.3 'Burst' Kinetics 9.4 Reversible Sequences of Reactions 9.5 Jump Kinetics 9.6 Sinusoidal Perturbations10 Estimation of Rate Constants 10.1 Value and Limitations of a Statistical Approach 10.2 Variance 10.3 Simple Linear Regression 10.4 Fitting the Michaelis-Menten Equation 10.5 Final Comments on the Double-Reciprocal Plot 10.6 Standard Errors of V and Km 10.7 General Linear Model and Applications to More Complex Cases 10.8 Some Difficulties in Fitting Data 10.9 Statistical Aspects of the Direct Linear Plot 10.10 Final NoteReferencesIndex