Sedimentation Equilibrium Analytical Ultracentrifugation

Auteur
Dr. Schuck obtained his Ph.D. from the Goethe-University Frankfurt am Main, Germany, where he worked on interactions of integral proteins of the erythrocyte membrane using analytical ultracentrifugation. He received his post-doctoral research training in physical biochemistry with Dr. Allen Minton at NIDDK, and joined the Bioengineering and Physical Science Program of NCRR as a Research Fellow in 1997. He is currently a Earl Stadtman Tenure-Track Investigator and Chief of the Dynamics of Macromoleular Assembly Section, Laboratory of Cellular Imaging and Macromolecular Biophysics at the National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health. Dr. Ghirlando earned his Ph.D. at the Weizmann Institute of Science, and is a Staff Scientist at the Laboratory of Molecular Biology, at the National Institute of Diabetes and Digestive and Kidney Diseases, NIH. His interest include the study of the in vivo chromatin structure at the 30-nm fiber level, and more generally, the development of hydrodynamic methodology for the study of challenging biomacromolecular interactions.

Résumé

A follow-up to the experimental and instrumental aspects described in Basic Principles of Analytical Ultracentrifugation, the volume Sedimentation Velocity Analytical Ultracentrifugation: Discrete Species and Size-Distributions of Macromolecules and Particles describes the theory and practice of data analysis. Mathematical models for the sedimentation process and the evolution of detected signals are developed in a comprehensive framework, jointly with the description of current and historical strategies for how to extract from noisy experimental data the physical parameters of interest, such as size, mass, and shape, composition, and polydispersity of sedimenting particles. The methods are extensively illustrated, and supported with practical applications, as well as cross-references where to find the methods in the public domain software SEDFIT and SEDPHAT. The systems covered are discrete or polydisperse mixtures of sedimenting molecules or particles in dilute solution, such as proteins and other biomolecules and their stable complexes, man-made polymers, and nanoparticles, observed in different optical systems. These methods form the essential foundation for the analysis of dynamic interacting systems, which are covered in a separate volume.

Contenu

Concepts of Modern Sedimentation Velocity Data Analysis. Sedimentation of Discrete Non-Interacting Particles. General Properties of Sedimentation Coefficient Distributions. Distributions of Non-Diffusing Particles. Distributions of Diffusing Particles. Sedimentation Coefficient Distributions from Boundary Derivatives and Extrapolations. Multi-Component Distributions. Practical Analysis of Non-Interacting Systems. Appendix: Numerical Methods. Bibliography.