Ultracentrifugation in Biochemistry

Ultracentrifugation in Biochemistry discusses the fundamental aspects of ultracentrifugation. The book begins with a sketch of the field, highlighting some of the principal developments. Following this is a chapter that discusses ultracentrifugation in general terms and describes the division of the field into three major areas.
The subsequent chapter deals with developments of the experimental aspects of the field such as improvements in the instrument itself, cells, rotors, measurement, and control of temperature, and the various optical systems. The remainder of the book discusses the fundamental principles of sedimentation velocity, transient states, and sedimentation equilibrium. A section is also included which deals with interpretation of sedimentation data in terms of hydrodynamic models, charge effects, and interactions in multicomponent systems. This book is likely to become an indispensable companion to the laboratory worker who is planning and conducting an ultracentrifuge run for almost any purpose. It should also be of fundamental value to the thoughtful student or investigator who wants to know the present state of knowledge in the field, both experimental and theoretical.

Foreword

Preface

I. Introduction

II. General Considerations

III. Experimental Aspects

1. The Electrically Driven Ultracentrifuge

2. The Magnetically Suspended Ultracentrifuge

3. Analytical Rotors

4. Temperature Measurement and the Stretching of Rotors

5. Analytical Cells

a. Double Sector Cell

b. Synthetic Boundary Cells

c. Separation Cells

d. Other Cells

6. Optical Methods

a. Lamm Scale Method

b. Schlieren Methods

c. Interference Methods

d. Light Absorption Methods

7. Convection-free Sedimentation

IV. Sedimentation Velocity

1. The Relationship between Boundary Movement and the Sedimentation Velocity of Individual Molecules

2. Radial Dilution

3. Measurement of Sedimentation Coefficients

a. Moving Boundary Method

b. The Transport Method

4. Dependence of Sedimentation Coefficient on Concentration

5. Differential Sedimentation Rates

6. Sedimentation of a Slow Component in the Presence of a Faster Species

7. Analysis of Mixtures of Several Sedimenting Components

8. Analysis of Boundaries in Terms of Homogeneity or Polydispersity

a. Spreading of Boundaries Due to Diffusion

b. Spreading of Boundaries Due to Polydispersity

c. Self-Sharpening of Boundaries Due to the Concentration Dependence of Sedimentation Coefficients

d. Distortion of the Boundary Due to the Johnston-Ogston Effect

e. General Considerations in the Demonstration of Homogeneity

9. Analysis of Systems of Reversibly Interacting Components

a. Association-Dissociation Equilibria Involving a Single Component

b. Interactions among Different Molecular Species

10. A Differential Method for the Direct Measurement of Small Differences in Sedimentation Coefficients

11. The Effect of Pressure on Sedimentation

V. The Transient States during the Approach to Sedimentation Equilibrium

1. Molecular Weight Determinations by the Archibald Method

2. Distributions throughout the Entire Cell

VI. Sedimentation Equilibrium

1. General Considerations

2. Two Component Systems

3. Charged Macromolecules

4. Multicomponent Systems and Sedimentation in a Density Gradient

VII. Interpretation of Sedimentation Data

1. The Svedberg Equation

2. The Effect of Solvation

3. Nonideal Solutions

4. Effect of Charge on Sedimentation

5. Multicomponent Systems

6. The Frictional Coefficient

References

Glossary of Terms

Author Index

Subject Index