Enzyme kinetics : a modern approach / Alejandro G. Marangoni.

Format:

Book

Author/Creator:

Marangoni, Alejandro G., 1965-

Language:

English

Subjects (All):

Enzyme kinetics.

Enzymes.

Kinetics.

Models, Chemical.

Medical Subjects:

Enzymes.

Kinetics.

Models, Chemical.

Physical Description:

xiv, 229 pages : illustrations ; 25 cm

Place of Publication:

Hoboken, N.J. : Wiley-Interscience, [2003]

Summary:

Practical Enzyme Kinetics provides a practical how-to guide for beginning students, technicians, and non-specialists for evaluating enzyme kinetics using common software packages to perform easy enzymatic analyses.

Contents:

1 Tools and Techniques of Kinetic Analysis 1

1.1 Generalities 1

1.2 Elementary Rate Laws 2

1.2.1 Rate Equation 2

1.2.2 Order of a Reaction 3

1.2.3 Rate Constant 4

1.2.4 Integrated Rate Equations 4

1.2.4.1 Zero-Order Integrated Rate Equation 4

1.2.4.2 First-Order Integrated Rate Equation 5

1.2.4.3 Second-Order Integrated Rate Equation 7

1.2.4.4 Third-Order Integrated Rate Equation 8

1.2.4.5 Higher-Order Reactions 9

1.2.4.6 Opposing Reactions 9

1.2.4.7 Reaction Half-Life 11

1.2.5 Experimental Determination of Reaction Order and Rate Constants 12

1.2.5.1 Differential Method (Initial Rate Method) 12

1.2.5.2 Integral Method 13

1.3 Dependence of Reaction Rates on Temperature 14

1.3.1 Theoretical Considerations 14

1.3.2 Energy of Activation 18

1.4 Acid-Base Chemical Catalysis 20

1.5 Theory of Reaction Rates 23

1.6 Complex Reaction Pathways 26

1.6.1 Numerical Integration and Regression 28

1.6.1.1 Numerical Integration 28

1.6.1.2 Least-Squares Minimization (Regression Analysis) 29

1.6.2 Exact Analytical Solution (Non-Steady-State Approximation) 39

1.6.3 Exact Analytical Solution (Steady-State Approximation) 40

2 How Do Enzymes Work? 41

3 Characterization of Enzyme Activity 44

3.1 Progress Curve and Determination of Reaction Velocity 44

3.2 Catalysis Models: Equilibrium and Steady State 48

3.2.1 Equilibrium Model 48

3.2.2 Steady-State Model 49

3.2.3 Plot of v versus [S] 50

3.3 General Strategy for Determination of the Catalytic Constants K[subscript m] and V[subscript max] 52

3.4 Practical Example 53

3.5 Determination of Enzyme Catalytic Parameters from the Progress Curve 58

4 Reversible Enzyme Inhibition 61

4.1 Competitive Inhibition 61

4.2 Uncompetitive Inhibition 62

4.3 Linear Mixed Inhibition 63

4.4 Noncompetitive Inhibition 64

4.5 Applications 65

4.5.1 Inhibition of Fumarase by Succinate 65

4.5.2 Inhibition of Pancreatic Carboxypeptidase A by [beta]-Phenylpropionate 67

4.5.3 Alternative Strategies 69

5 Irreversible Enzyme Inhibition 70

5.1 Simple Irreversible Inhibition 72

5.2 Simple Irreversible Inhibition in the Presence of Substrate 73

5.3 Time-Dependent Simple Irreversible Inhibition 75

5.4 Time-Dependent Simple Irreversible Inhibition in the Presence of Substrate 76

5.5 Differentiation Between Time-Dependent and Time-Independent Inhibition 78

6 pH Dependence of Enzyme-Catalyzed Reactions 79

6.1 The Model 79

6.2 pH Dependence of the Catalytic Parameters 82

6.3 New Method of Determining pK Values of Catalytically Relevant Functional Groups 84

7 Two-Substrate Reactions 90

7.1 Random-Sequential Bi Bi Mechanism 91

7.1.1 Constant [A] 93

7.1.2 Constant [B] 93

7.2 Ordered-Sequential Bi Bi Mechanism 95

7.2.1 Constant [B] 95

7.2.2 Constant [A] 96

7.2.3 Order of Substrate Binding 97

7.3 Ping-Pong Bi Bi Mechanism 98

7.3.1 Constant [B] 99

7.3.2 Constant [A] 99

7.4 Differentiation Between Mechanisms 100

8 Multisite and Cooperative Enzymes 102

8.1 Sequential Interaction Model 103

8.1.1 Basic Postulates 103

8.1.2 Interaction Factors 105

8.1.3 Microscopic versus Macroscopic Dissociation Constants 106

8.1.4 Generalization of the Model 107

8.2 Concerted Transition or Symmetry Model 109

8.3 Application 114

8.4 Reality Check 115

9 Immobilized Enzymes 116

9.1 Batch Reactors 116

9.2 Plug-Flow Reactors 118

9.3 Continuous-Stirred Reactors 119

10 Interfacial Enzymes 121

10.1 The Model 122

10.1.1 Interfacial Binding 122

10.1.2 Interfacial Catalysis 123

10.2 Determination of Interfacial Area per Unit Volume 125

10.3 Determination of Saturation Interfacial Enzyme Coverage 127

11 Transient Phases of Enzymatic Reactions 129

11.1 Rapid Reaction Techniques 130

11.2 Reaction Mechanisms 132

11.2.1 Early Stages of the Reaction 134

11.2.2 Late Stages of the Reaction 135

11.3 Relaxation Techniques 135

12 Characterization of Enzyme Stability 140

12.1 Kinetic Treatment 140

12.1.1 The Model 140

12.1.2 Half-Life 142

12.1.3 Decimal Reduction Time 143

12.1.4 Energy of Activation 144

12.1.5 Z Value 145

12.2 Thermodynamic Treatment 146

12.3.1 Thermodynamic Characterization of Stability 151

12.3.2 Kinetic Characterization of Stability 156

13 Mechanism-Based Inhibition / Leslie J. Copp 158

13.1 Alternate Substrate Inhibition 159

13.2 Suicide Inhibition 163

13.3.1 Alternative Substrate Inhibition 169

13.3.2 Suicide Inhibition 170

14 Putting Kinetic Principles into Practice / Kirk L. Parkin 174

14.1 Were Initial Velocities Measured? 175

14.2 Does the Michaelis

Menten Model Fit? 177

14.3 What Does the Original [S] versus Velocity Plot Look Like? 179

14.4 Was the Appropriate [S] Range Used? 181

14.5 Is There Consistency Working Within the Context of a Kinetic Model? 184

15 Use of Enzyme Kinetic Data in the Study of Structure

Function Relationships of Proteins / Takuji Tanaka, Rickey Y. Yada 193

15.1 Are Proteins Expressed Using Various Microbial Systems Similar to the Native Proteins? 193

15.2 What Is the Mechanism of Conversion of a Zymogen to an Active Enzyme? 195

15.3 What Role Does the Prosegment Play in the Activation and Structure

Function of the Active Enzyme? 198

15.4 What Role Do Specific Structures and/or Residues Play in the Structure

Function of Enzymes? 202

15.5 Can Mutations be Made to Stabilize the Structure of an Enzyme to Environmental Conditions? 205

15.5.1 Charge Distribution 205

15.5.2 N-Frag Mutant 208

15.5.3 Disulfide Linkages 210.

Notes:

Includes bibliographical references (pages 217-219) and index.

ISBN:

0471159859

OCLC:

50554780