Andrea Bellelli, Jannette Carey

Reversible Ligand Binding (eBook, PDF)

Theory and Experiment

• Format: PDF

Produktbeschreibung

Presents the physical background of ligand binding and instructs on how experiments should be designed and analyzed Reversible Ligand Binding: Theory and Experiment discusses the physical background of protein-ligand interactions--providing a comprehensive view of the various biochemical considerations that govern reversible, as well as irreversible, ligand binding. Special consideration is devoted to enzymology, a field usually treated separately from ligand binding, but actually governed by identical thermodynamic relationships. Attention is given to the design of the experiment, which aids in showing clear evidence of biochemical features that may otherwise escape notice. Classical experiments are reviewed in order to further highlight the importance of the design of the experiment. Overall, the book supplies students with the understanding that is necessary for interpreting ligand binding experiments, formulating plausible reaction schemes, and analyzing the data according to the chosen model(s). Topics covered include: theory of ligand binding to monomeric proteins; practical considerations and commonly encountered problems; oligomeric proteins with multiple binding sites; ligand binding kinetics; hemoglobin and its ligands; single-substrate enzymes and their inhibitors; two-substrate enzymes and their inhibitors; and rapid kinetic methods for studying enzyme reactions. * Bridges theory of ligand binding and allostery with experiments * Applies historical and physical insight to provide a clear understanding of ligand binding * Written by a renowned author with long-standing research and teaching expertise in the area of ligand binding and allostery * Based on FEBS Advanced Course lectures on the topic Reversible Ligand Binding: Theory and Experiment is an ideal text reference for students and scientists involved in biophysical chemistry, physical biochemistry, biophysics, molecular biology, protein engineering, drug design, pharmacology, physiology, biotechnology, and bioengineering.

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Produktdetails

• Verlag: John Wiley & Sons
• Seitenzahl: 304
• Erscheinungstermin: 24. Oktober 2017
• Englisch
• ISBN-13: 9781119238478
• Artikelnr.: 52560674

Autorenporträt

Andrea Bellelli, PhD is a Professor of Biochemistry at the University of Rome Sapienza. He chaired the Department of Biochemical Sciences "A. Rossi Fanelli" and currently chairs the Medicine and Surgery "B" school at the same University. His research focuses on structural and functional properties of oxygen carrying proteins. Jannette Carey, PhD is a Professor of Chemistry at Princeton University and a visiting scientist of the Academy of Sciences of the Czech Republic at Nové Hrady, where she initiated and organizes a biennial FEBS practical and lecture course, Ligand binding theory and practice.

Inhaltsangabe

Preface xi

Acknowledgments xiii

Part I Ligand Binding to Single Binding Site Targets 1

1 Theory of Ligand Binding to Monomeric Proteins 3

1.1 Importance of Ligand?]Binding Phenomena in Biology 3

1.2 Preliminary Requirements for Ligand?]Binding Study 5

1.3 Chemical Equilibrium and the Law of Mass Action 5

1.4 The Hyperbolic and Sigmoidal Representations of the Ligand?]Binding Isotherms 7

1.5 The Important Concept of X1/2 11

1.6 Other Representations of the Ligand?]Binding Isotherm 11

1.7 Effect of Temperature: Thermodynamic Relationships 14

1.8 Replacement Reactions: Competitive Ligands 17

1.9 Heterotropic Linkage: Non?]Competitive Binding of Two Ligands 20

1.10 Allostery and Allosteric Phenomena in Monomeric Proteins 23

1.11 The Special Case of Cys Ligands (and Similar Reactions) 24

1.12 Other Special Cases 27

2 Ligand?]Binding Kinetics for Single?]Site Proteins 31

2.1 Basic Concepts of Chemical Kinetics: Irreversible Reactions 31

2.2 Reversible Reactions: Equilibrium and Kinetics 35

2.3 More Complex Kinetic Mechanisms 37

2.4 Reactions with Molecularity Higher Than Two 40

2.5 Classical Methods for the Study of Ligand?]Binding Kinetics 41

2.6 Photochemical Kinetic Methods 44

2.7 The Kinetics of Replacement Reactions 47

Appendix to Chapter 2: Principles of Data Analysis 51

3 Practical Considerations and Commonly Encountered Problems 53

3.1 Design of the Experiment: The Free Ligand Concentration 53

3.2 The Signal and the Concentration of the Target 56

3.3 Test of the Reversibility of the Reaction 59

3.4 Frequent Abuses of the Concept of X1/2 60

3.5 Two Common Problems: Protein Precipitation and Baseline Shifts 62

3.6 Low?]Affinity Ligands 63

3.7 High?]Affinity Ligands 65

3.8 Determination of Binding Stoichiometry 67

3.9 Ligands Occupying a Thermodynamic Phase Different from the Protein 69

3.10 Mixtures of Isoforms 71

3.11 Poor or Absent Signal 73

Part II Ligand Binding to Multiple Binding Site Proteins 75

4 Proteins with Multiple Binding Sites 77

4.1 Multiple Binding Sites: Determination of the Binding Stoichiometry 77

4.2 The Binding Polynomial of a Homooligomeric Protein Made Up of Identical Subunits 79

4.3 Intramolecular Heterogeneity 84

4.4 Oligomeric Proteins with Interacting Binding Events: Homotropic Linkage 86

4.5 Cooperativity: Biochemistry and Physiology 91

4.6 Allostery and Symmetry: The Allosteric Model of Cooperativity 94

4.7 Two Alternative Concepts of Cooperativity 100

4.8 Ligand Replacement in Oligomeric Proteins 104

4.9 Heterotropic Linkage in Multimeric Proteins 105

4.10 Heterotropic Linkage and the Allosteric Model 110

Appendix 4.1 Statistical Distribution of the Ligand Among the Binding Sites: Statistical Factors 112

Appendix 4.2 Symmetry of the X Versus Log([X]) Plot: The Concept of Xm 113

5 Ligand?]Linked Association and Dissociation 117

5.1 Quaternary Constraint and Quaternary Enhancement 118

5.2 The Reversibly Dissociating Homodimer Devoid of Ligand?]Linked Association Equilibria 119

5.3 Ligand?]Linked Association?]Dissociation in the Non?]Cooperative Homodimer 122

5.4 Oligomers That Dissociate Into Monomers Upon Ligand Binding 126

5.5 Monomers That Self?]Associate to Homodimers Upon Ligation 129

5.6 Ligand?]Linked Association?]Dissociation in Cooperative Proteins 130