Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2008 May:Chapter 18:18.15.1-18.15.39.
doi: 10.1002/0471142735.im1815s81.

Characterizing protein-protein interactions by sedimentation velocity analytical ultracentrifugation

Patrick H Brown  1 , Andrea Balbo  1 , Peter Schuck  1
Affiliations

Characterizing protein-protein interactions by sedimentation velocity analytical ultracentrifugation

Patrick H Brown et al. Curr Protoc Immunol. 2008 May.

Abstract

This unit introduces the basic principles and practice of sedimentation velocity analytical ultracentrifugation for the study of reversible protein interactions, such as the characterization of self-association, heterogeneous association, multi-protein complexes, binding stoichiometry, and the determination of association constants. The analytical tools described include sedimentation coefficient and molar mass distributions, multi-signal sedimentation coefficient distributions, Gilbert-Jenkins theory, different forms of isotherms, and global Lamm equation modeling. Concepts for the experimental design are discussed, and a detailed step-by-step protocol guiding the reader through the experiment and the data analysis is available as an Internet resource.

PubMed Disclaimer

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

Literature Cited

References
    1. Ali, S.A., Iwabuchi, N., Matsui, T., Hirota, K., Kidokoro, S., Arai, M., Kuwajima, K., Schuck, P., and Arisaka, F. 2003. Rapid and dynamic association equilibrium of a molecular chaperone, gp57A, of bacteriophage T4. Biophys. J. 85: 2606-2618.
    1. Arthos, J., Cicala, C., Steenbeke, T.D., Chun, T.-W., Dela Cruz, C., Hanback, D.B., Khazanie, P., Nam, D., Schuck, P., Selig, S.M., Van Ryk, D., Chaikin, M.A., and Fauci, A.S. 2002. Biochemical and biological characterization of a dodecameric CD4-Ig fusion protein: Implications for therapeutic and vaccine strategies. J. Biol. Chem. 277: 11456-11464.
    1. Balbo, A. and Schuck, P. 2005. Analytical ultracentrifugation in the study of protein self-association and heterogeneous protein-protein interactions. In Protein-Protein Interactions. (E. Golemis and P.D. Adams, eds.) pp. 253-277. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
    1. Balbo, A., Minor, K.H., Velikovsky, C.A., Mariuzza, R., Peterson, C.B., and Schuck, P. 2005. Studying multi-protein complexes by multi-signal sedimentation velocity analytical ultracentrifugation. Proc. Natl. Acad. Sci. U.S.A. 102: 81-86.
    1. Berkowitz, S.A. 2006. Role of analytical ultracentrifugation in assessing the aggregation of protein biopharmaceuticals. AAPS J. 8: E590-E605.
Key References
General reviews
    1. Balbo and Schuck, 2005. See above.
    1. Howlett et al., 2006. See above.
    1. Laue, T. 2001. Biophysical studies by ultracentrifugation. Curr. Opin. Struct. Biol. 11: 579-583.
    1. Lebowitz et al., 2002. See above.
    1. Schachman, 1959. See above.
References on special topics
    1. Berkowitz, S.A. and Philo, J.S. 2006. Monitoring the homogeneity of adenovirus preparations (a gene therapy delivery system) using analytical ultracentrifugation. Anal. Biochem. 362: 16-37.
    1. Durchschlag, H. 1986. Specific volumes of biological macromolecules and some other molecules of biological interest. In Thermodynamic Data for Biochemistry and Biotechnology. (H.-J. Hinz, ed.) pp. 45-128. Springer, Berlin.
    1. Gohon et al., 2004. See above.
    1. Howlett, G.J. 1992. Sedimentation analysis of membrane proteins. In Analytical Ultracentrifugation in Biochemistry and Polymer Science (S.E. Harding, A.J. Rowe, and J.C. Horton, eds.) pp. 470-483. The Royal Society of Chemistry, Cambridge.
    1. Lebowitz, J., Teale, M., and Schuck, P.W. 1998. Analytical band centrifugation of proteins and protein complexes. Biochem. Soc. Trans. 26: 745-749.
Modern SV data analysis for interacting systems
    1. Balbo et al., 2005. See above.
    1. Dam and Schuck, 2005. See above.
    1. Dam et al., 2005. See above.
    1. Schuck, 2003. See above.
Internet Resources
    1. http://www.analyticalultracentrifugation.com/SVProtocols.htm
    1. http://www.analyticalultracentrifugation.com
    1. http://www.analyticalultracentrifugation.com/sedphat/sedphat.htm

LinkOut - more resources