Enzymes : a practical introduction to structure, mechanism, and data analysis / Robert A. Copeland.

Format:

Book

Author/Creator:

Copeland, Robert Allen.

Contributor:

Dr. William McDowell Mastin Fund.

Language:

English

Subjects (All):

Enzymes.

Enzymology.

Medical Subjects:

Enzymes.

Physical Description:

xvi, 397 pages : illustrations ; 25 cm

Edition:

Second edition.

Place of Publication:

New York : J. Wiley-VCH, [2000]

Summary:

Supplemented with appendices providing contact information for suppliers of reagents and equipment for enzyme studies, as well as a survey of useful Internet sites and computer software for enzymatic data analysis, Enzymes, Second Edition is the ultimate practical guide for scientists and students in biochemical, pharmaceutical, biotechnical, medicinal, and agricultural/food-related research.

Contents:

1 A Brief History of Enzymology 1

1.1 Enzymes in Antiquity 2

1.2 Early Enzymology 3

1.3 The Development of Mechanistic Enzymology 4

1.4 Studies of Enzyme Structure 5

1.5 Enzymology Today 7

2 Chemical Bonds and Reactions in Biochemistry 11

2.1 Atomic and Molecular Orbitals 11

2.2 Thermodynamics of Chemical Reactions 23

2.3 Acid-Base Chemistry 29

2.4 Noncovalent Interactions in Reversible Binding 32

2.5 Rates of Chemical Reactions 35

3 Structural Components of Enzymes 42

3.1 The Amino Acids 42

3.2 The Peptide Bond 53

3.3 Amino Acid Sequence or Primary Structure 55

3.4 Secondary Structure 57

3.5 Tertiary Structure 62

3.6 Subunits and Quaternary Structure 65

3.7 Cofactors in Enzymes 68

4 Protein-Ligand Binding Equilibria 76

4.1 The Equilibrium Dissociation Constant, K[subscript d] 76

4.2 The Kinetic Approach to Equilibrium 78

4.3 Binding Measurements at Equilibrium 80

4.4 Graphic Analysis of Equilibrium Ligand Binding Data 88

4.5 Equilibrium Binding with Ligand Depletion (Tight Binding Interactions) 94

4.6 Competition Among Ligands for a Common Binding Site 95

4.7 Experimental Methods for Measuring Ligand Binding 96

5 Kinetics of Single-Substrate Enzyme Reactions 109

5.1 The Time Course of Enzymatic Reactions 109

5.2 Effects of Substrate Concentration on Velocity 111

5.3 The Rapid Equilibrium Model of Enzyme Kinetics 113

5.4 The Steady State Model of Enzyme Kinetics 115

5.5 The Significance of k[subscript cat] and K[subscript m] 120

5.6 Experimental Measurement of k[subscript cat] and K[subscript m] 124

5.7 Other Linear Transformations of Enzyme Kinetic Data 133

5.8 Measurements at Low Substrate Concentrations 136

5.9 Deviations from Hyperbolic Kinetics 137

5.10 Transient State Kinetic Measurements 141

6 Chemical Mechanisms in Enzyme Catalysis 146

6.1 Substrate-Active Site Complementarity 147

6.2 Rate Enhancement Through Transition State Stabilization 151

6.3 Chemical Mechanisms for Transition State Stabilization 154

6.4 The Serine Proteases: An Illustrative Example 178

6.5 Enzymatic Reaction Nomenclature 184

7 Experimental Measures of Enzyme Activity 188

7.1 Initial Velocity Measurements 188

7.2 Detection Methods 204

7.3 Separation Methods in Enzyme Assays 223

7.4 Factors Affecting the Velocity of Enzymatic Reactions 238

7.5 Reporting Enzyme Activity Data 257

7.6 Enzyme Stability 258

8 Reversible Inhibitors 266

8.1 Equilibrium Treatment of Reversible Inhibition 268

8.2 Modes of Reversible Inhibition 270

8.3 Graphic Determination of Inhibitor Type 273

8.4 Dose-Response Curves of Enzyme Inhibition 282

8.5 Mutually Exclusive Binding of Two Inhibitors 287

8.6 Structure-Activity Relationships and Inhibitor Design 291

9 Tight Binding Inhibitors 305

9.1 Identifying Tight Binding Inhibition 305

9.2 Distinguishing Inhibitor Type for Tight Binding Inhibitors 307

9.3 Determining K[subscript i] for Tight Binding Inhibitors 310

9.4 Use of Tight Binding Inhibitors to Determine Active Enzyme Concentration 313

10 Time-Dependent Inhibition 318

10.1 Progress Curves for Slow Binding Inhibitors 321

10.2 Distinguishing Between Slow Binding Schemes 325

10.3 Distinguishing Between Modes of Inhibitor Interaction with Enzyme 330

10.4 Determining Reversibility 332

10.5 Examples of Slow Binding Enzyme Inhibitors 334

11 Enzyme Reactions with Multiple Substrates 350

11.1 Reaction Nomenclature 350

11.2 Bi Bi Reaction Mechanisms 352

11.3 Distinguishing Between Random and Compulsory Ordered Mechanisms by Inhibition Pattern 357

11.4 Isotope Exchange Studies for Distinguishing Reaction Mechanisms 360

11.5 Using the King-Altman Method to Determine Velocity Equations 362

12 Cooperativity in Enzyme Catalysis 367

12.1 Historic Examples of Cooperativity and Allostery in Proteins 368

12.2 Models of Allosteric Behavior 373

12.3 Effects of Cooperativity on Velocity Curves 379

12.4 Sigmoidal Kinetics for Nonallosteric Enzymes 382.

Notes:

"Published simultaneously in Canada."

Includes bibliographical references and index.

Local Notes:

Acquired for the Penn Libraries with assistance from the Dr. William McDowell Mastin Fund.

ISBN:

0471359297

OCLC:

42619660