Multiple Equilibria in Proteins

Multiple Equilibria in Proteins covers the multiple interactions between small ions and molecules and a protein molecule. The book also deals with the physicochemical mechanisms of this interaction and the information about protein structure and the forces stabilizing that structure. The text discusses the mathematical description of complex formation, the thermodynamic analysis of binding data, and various theoretical models which can be used to describe the phenomena of small molecule-macromolecule interactions. The measurement of complex formation; the binding of neutral molecules; and hydrogen-ion equilibria are also considered. The book further tackles metal-ion binding; the binding of organic ions by proteins; as well as protein-protein interaction. Chemists and biochemists will find the book useful.

Preface

I. Introduction

References

II. Thermodynamics and Model Systems

I. Introduction

II. Some Concepts in Thermodynamics

III. Multiple Binding without Interaction between Sites

IV. Binding with Interaction between Sites

A. Debye-Hückel-Born Treatment of Electrostatic Interaction

B. Tanford-Kirkwood Model for Electrostatic Interaction

C. Hill Model for Cylindrical Rods

V. Method of Data Treatment

VI. Thermodynamics of Multiple Equilibria

VII. Binding-Induced Phase Transitions

VIII. Linked Functions

References

III. The Measurement of Complex Formation

I. Introduction

II. Subtractive Methods

A. EMF Measurements of Free Ligand Activity

B. Equilibrium Dialysis

C. Ultrafiltration and Ultracentrifugation

D. Gel Filtration

E. Partition Equilibrium between Phases

F. Electrical Conductivity

G. Polarography

H. Determination of Free Ligand without Separation of Phases

III. Direct Measurements

A. Changes in Light Absorption of Ligand

B. Fluorescence Methods

C. Optical Rotation

D. Nuclear Magnetic Resonance

E. X-Ray Diffraction

F. Refractive Index

G. Light-Scattering and Osmotic Pressure

H. Biological Activity

IV. Electrostatic Methods

A. Electrophoresis

B. Changes in the Titration Curves of Proteins due to the Presence of Bound Ions

C. The pH Method of Scatchard and Black—Displacement of pH of Isoionic Solutions by Added Salt

D. Dielectric Increment and Dispersion

V. Other Methods

A. Complement Fixation

B. Methods which Depend on Competition

References

IV. Binding of Neutral Molecules

I. Introduction

II. Results Obtained with Particular Proteins

A. Serum Albumins

B. β-Lactoglobulin

C. Myoglobin and Hemoglobin

D. Lysozyme

E. Other Enzymes

F. Globulins

G. α-Acid Glycoprotein (Human)

H. Concanavalin A

I. Myosin

III. The Hydration of Proteins

A. The State of Combined Water

IV. Interactions with Other Solvents

A. Dependence on High Powers of Concentration

V. Mechanism of Binding of Neutral Molecules

References

V. Hydrogen-Ion Equilibria

I. Introduction

II. Prototropic Groups in Proteins

III. Some Experimental Details

A. Isoionic Point

B. Definition of pH

IV. An Experimental Titration Curve

V. Anomalies Illustrated by Specific Proteins

A. Bovine Serum Albumin—Abnormal Carboxylate Groups

B. ß-Lactoglobulin—Abnormal Carboxyl and Thiol Groups

C. Ribonuclease—Abnormal Phenolic Groups

D. Hemoglobin—Abnormal Imidazole Groups

E. Insulin and Zinc Insulin

F. Paramyosin, a Rodlike Protein

VI. Hydrogen-Ion Equilibria of Denatured Proteins

VII. Unfolding as a Function of Protonation

A. Bovine Serum Albumin

B. E. coli Alkaline Phosphatase

VIII. Summary

References

VI. Metal-Ion Binding

I. Metal Ions and Their Complexes in Solution

II. Experimental Methods and Data Treatment

III. Proteins Containing Metal Ions Necessary for Biological Activity

IV. Protein-Metal Complex Formation

A. Bovine Serum Albumin

B. Sperm Whale Myoglobin

C. Ribonuclease

D. Vasopressin and Oxytocin

E. Insulin

F. Membranes

G. Ribosomes

H. S-100, a Protein of the Nervous System

References

VII. Binding of Organic Ions by Proteins

I. Introduction

II. Large Organic Ions—Ionic Detergents

A. Early Work on Detergent Anions

B. Isotherms of Anionic Detergents

C. The Binding of Fatty Acids Compared to Other Ligands. Nature of the Binding Site

D. Criteria of Unfolding. Viscosity Deuterium Exchange, ORD, and Difference Spectra

E. Effects of pH on Anion Binding

F. Other Effects of Detergents on Proteins

G. Enthalpy Change in Binding. Uniqueness of Two Highest Energy Sites of BSA

H. Binding of Polyions

III. Dyes, Dye-Like, and Other Cyclic Molecules Binding as Ions

A. Early Work

B. The Affinities of Some Dyes to Proteins

C. Fluorescent Ligands

D. Dye Interactions with Denatured Protein

E. Protection by Dyes against Denaturation

F. Phytic Acid

IV. Binding of Small Nonaromatic Ions to Proteins

A. Early Work

B. Thermodynamic Studies (Affinities and Numbers of Sites)

C. Supporting Data and Applications

D. Additional Data

E. Cations

F. Evidence from Electrophoresis

G. Evidence from Conformational Effects

H. Protection against Unfolding

I. Miscellaneous Reports of Small Ion Binding to Proteins

J. Kinetics of Binding Processes

K. Effect of Binding on Subunit Interactions

References

VIII. Protein-Protein Interaction

I. Introduction

II. Quarternary Structure in Proteins

III. Glutamic Dehydrogenase

IV. Tobacco Mosaic Virus Protein

V. Hemerythrin

References

IX. Summary and Conclusions

References

Author Index

Subject Index