Cornish-Bowden Fundamentals of Enzyme Kinetics

1. Auflage 2014
ISBN: 978-1-4831-6119-8
Verlag: Elsevier Science & Techn.
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, 244 Seiten, Web PDF

Fundamentals of Enzyme Kinetics details the rate of reactions catalyzed by different enzymes and the effects of varying the conditions on them. The book includes the basic principles of chemical kinetics, especially the order of a reaction and its rate constraints. The text also gives an introduction to enzyme kinetics - the idea of an enzyme-substrate complex; the Michaelis-Menten equation; the steady state treatment; and the validity of its assumption. Practical considerations, the derivation of steady-state rate equations, inhibitors and activators, and two-substrate reactions are also explained. Problems after the end of each chapter have also been added, as well as their solutions at the end of the book, to test the readers' learning. The text is highly recommended for undergraduate students in biochemistry who wish to study about enzymes or focus completely on enzymology, as most of the mathematics used in this book, which have been explained in detail to remove most barriers of understanding, is elementary.

Autoren/Hrsg.

Cornish-Bowden, Athel

Inhaltsverzeichnis

1;Cover
;1
2;Fundamentals of Enzyme Kinetics;4
3;Copyright;5
4;Table of contents
;8
5;Preface;6
6;Principal symbols used in this book;12
7;Chapter 1.
Basic principles of chemical kinetics;16
7.1;1.1 Order of a reaction;16
7.2;1.2 Determination of the order of a reaction;19
7.3;1.3 Dimensions of rate constants;20
7.4;1.4
Reversible reactions;22
7.5;1.5 Determination of first-order rate constants;23
7.6;1.6 Influence of temperature on rate constants;24
7.7;1.7 Transition-state theory;27
7.8;Problems;29
8;Chapter 2. Introduction to enzyme kinetics;31
8.1;2.1 Early studies: the idea of an enzyme-substrate complex;31
8.2;2.2 Michaelis-Menten equation;32
8.3;2.3 Steady-state treatment;35
8.4;2.4 Validity of the steady-state assumption;39
8.5;2.5 Graphical representation of the Michaelis-Menten equation;40
8.6;2.6 Reversible Michaelis-Menten mechanism;45
8.7;2.7 Product inhibition;48
8.8;2.8 Integrated Michaelis-Menten equation;49
8.9;Problems;52
9;Chapter 3. Practical considerations;54
9.1;3.1 Purification of enzymes;54
9.2;3.2 Enzyme assays;55
9.3;3.3 Coupled assays;57
9.4;3.4 Protein determination;61
9.5;3.5 Presentation of results of a purification;62
9.6;3.6 Detecting enzyme inactivation;64
9.7;3.7 Experimental design: choice of substrate concentrations;66
9.8;3.8 Choice of pH, temperature and other conditions;68
9.9;3.9 Use of replicate observations;68
9.10;3.10 Treatment of ionic equilibria;70
9.11;Problems;73
10;Chapter 4. How to derive steady-state rate equations;75
10.1;4.1 King-Altman method;75
10.2;4.2 Modifications to the King-Altman method;79
10.3;4.3 Cha's method for reactions containing steps at equilibrium;82
10.4;4.4 Analysing mechanisms by inspection;84
10.5;Problems;86
11;Chapter 5. Inhibitors and activators;88
11.1;5.1 Reversible and irreversible inhibitors;88
11.2;5.2 Competitive inhibition;89
11.3;5.3 Mixed inhibition;91
11.4;5.4 Uncompetitive inhibition;93
11.5;5.5 Plotting inhibition results;93
11.6;5.6 Inhibition by a competing substrate;97
11.7;5.7 Activation;100
11.8;5.8 Hyperbolic inhibition and activation;103
11.9;5.9 Design of inhibition experiments
;104
11.10;5.10 Non-productive binding;106
11.11;5.11 Substrate inhibition;108
11.12;5.12 Chemical modification as a means of identifying essential groups;109
11.13;Problems;111
12;Chapter 6. Two-substrate reactions;114
12.1;6.1 Introduction;114
12.2;6.2 Types of mechanism;115
12.3;6.3 Rate equations;119
12.4;6.4 Initial-velocity measurements in the absence of products;124
12.5;6.5 Substrate inhibition;129
12.6;6.6 Product inhibition;131
12.7;6.7 Design of experiments;134
12.8;6.8 Isotope exchange;135
12.9;6.9 Reactions with three or more substrates;139
12.10;Problems;142
13;Chapter 7. Effects of pH and temperature on enzymes;145
13.1;7.1 pH and enzyme kinetics;145
13.2;7.2 Acid-base properties of proteins;146
13.3;7.3 Ionization of a dibasic acid;147
13.4;7.4 Effect of pH on enzyme kinetic constants;151
13.5;7.5 Ionization of the substrate;155
13.6;7.6 More complex pH effects;156
13.7;7.7 Temperature dependence of enzyme-catalysed reactions;156
13.8;Problems;160
14;Chapter 8. Control of enzyme activity;162
14.1;8.1 Necessity for metabolic control;162
14.2;8.2 Binding of oxygen to haemoglobin;164
14.3;8.3 Hill equation;167
14.4;8.4 Adair equation;169
14.5;8.5 Definition of co-operativity;172
14.6;8.6 Induced fit;174
14.7;8.7 Symmetry model of Monod, Wyman and Changeux;176
14.8;8.8 Sequential model of Koshland, Némethy and Filmer;181
14.9;8.9 Other models for co-operativity at equilibrium;186
14.10;8.10 Kinetic models of co-operativity;187
14.11;Problems;190
15;Chapter 9. Fast reactions;192
15.1;9.1 Limitations of steady-state measurements;192
15.2;9.2 Active-site titration: 'burst'kinetics;194
15.3;9.3 Flow methods;198
15.4;9.4 Relaxation methods;203
15.5;9.5 Transient-state kinetics of systems far from equilibrium;204
15.6;9.6 Simplification of complex mechanisms;208
15.7;9.7 Kinetics of systems close to equilibrium;211
15.8;Problems;213
16;Chapter 10. Estimation of kinetic constants;215
16.1;10.1 Cautionary note;215
16.2;10.2 Least-squares fit to the Michaelis-Menten equation;216
16.3;10.3 Statistical aspects of the direct linear plot;218
16.4;10.4 Precision of Km and V estimates;222
16.5;10.5 Examination of residuals;223
16.6;Problems;226
17;References;227
18;Solutions to problems;234
19;Index;238