Refining Hydrodynamic Shapes of Proteins: The Combination of Data From Analytical Ultracentrifugation and Time-Resolved Fluorescence Anisotropy Decay
Classical physical biochemical techniques are currently experiencing a renaissance as the result of several technological advances made over the past decade. One major reason for this renaissance is the need to understand the structures and functional characteristics of wild-type and mutant proteins that are now readily available through recombinant-DNA methods. Mutations of interest include alterations at specific functional sites, truncations, and switched domains. Another major reason for this renaissance is the availability of cheap, accessible computing power to facilitate data reduction. It is now possible and appropriate to combine the data from different physical techniques to obtain information which would not be obtainable from any single technique alone. This chapter examines a way in which the combination of analytical ultracentrifugation and time-resolved fluorescence anisotropy data permits knowledge of the hydrodynamic shape of a protein to be refined.
KeywordsFluorescence Anisotropy Rotational Correlation Time Analytical Ultracentrifugation Hydrated Sphere Partial Specific Volume
Unable to display preview. Download preview PDF.
- Cohn, E. J., & Edsall, J. T. (1943) Proteins, Amino Acids and Peptides as Ions and Dipolar Ions, p. 157, Rheinhold, New York, NY.Google Scholar
- Lakowicz, J. R. (1983) Principles of Fluorescence Spectroscopy, pp. 161–162, Plenum Press, New York, NY.Google Scholar
- Laue, T. M., Shah, B. D., Ridgeway, T. M., and Pelletier, S. M. (1992) Analytical Ultracentrifugation in Biochemistry and Polymer Science, pp. 90–125, Eds. S. Harding & A. Rowe, Royal Society of Chemistry, London.Google Scholar