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. 2007 Nov:Chapter 18:18.8.1-18.8.28.
doi: 10.1002/0471142735.im1808s79.

Measuring protein-protein interactions by equilibrium sedimentation

Andrea Balbo  1 , Patrick H Brown  1 , Emory H Braswell  2 , Peter Schuck  2
Affiliations

Measuring protein-protein interactions by equilibrium sedimentation

Andrea Balbo et al. Curr Protoc Immunol. 2007 Nov.

Abstract

This unit describes basic principles and practice of sedimentation equilibrium analytical ultracentrifugation for the study of reversible protein interactions, such as the characterization of self-association, heterogeneous association, and binding stoichiometry, as well as the determination of association constants. Advanced tools such as mass conservation analysis, multiwavelength analysis, and global analysis are introduced and discussed in the context of the experimental design. A detailed protocol guiding the investigator through the experimental steps and the data analysis is available as an internet resource.

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References

Literature Cited

References
    1. Adams, E.T., Jr., and Lewis, M.S. 1968. Sedimentation equilibrium in reacting systems. VI. Some applications to indefinite self-associations: Studies with beta-lactoglobulin A. Biochemistry 7:1044-1053.
    1. Advant, S.J., Braswell, E.H., Kumar, C.V., and Kalonia, D.S. 1995. The effect of pH and temperature on the self-association of recombinant human interleukin-2 as studied by equilibrium sedimentation. Pharm. Res. 12:637-641.
    1. Ansevin, A.T., Roark, D.E., and Yphantis, D.A. 1970. Improved ultracentrifuge cells for high-speed sedimentation equilibrium studies with interference optics. Anal. Biochem. 34:237-261.
    1. Arisaka, F. 1999. Applications and future perspectives of analytical ultracentrifugation. Tanpakushitsu Kakusan Koso 44:82-91.
    1. Arkin, M. and Lear, J.D. 2001. A new data analysis method to determine binding constants of small molecules using equilibrium analytical ultracentrifugation with absorption optics. Anal. Biochem. 299:98-107.
Key References
    1. Balbo and Schuck, 2005. See above.
    1. Howlett et al., 2006. See above.
    1. Lebowitz et al., 2002. See above.
    1. Schachman, 1959. See above.
    1. Schuck, 2007. See above.
Internet Resources
    1. http://www.nibib.nih.gov/Research/Intramural/LBPS/PB/auc
    1. http://www.analyticalultracentrifugation.com/sedphat/sedphat.htm
    1. http://www.analyticalultracentrifugation.com/sedphat/references.htm
    1. http://www.analyticalultracentrifugation.com/SEProtocols.htm

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