Unit

UNIT 18.15 Characterizing Protein-Protein Interactions by Sedimentation Velocity Analytical Ultracentrifugation

  1. Patrick H. Brown,
  2. Andrea Balbo,
  3. Peter Schuck

Published Online: 1 MAY 2008

DOI: 10.1002/0471142735.im1815s81

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Brown, P. H., Balbo, A. and Schuck, P. 2008. Characterizing Protein-Protein Interactions by Sedimentation Velocity Analytical Ultracentrifugation. Current Protocols in Immunology. 81:18.15:18.15.1–18.15.39.

Author Information

  1. National Institutes of Health, Bethesda, Maryland

Publication History

  1. Published Online: 1 MAY 2008
  2. Published Print: MAY 2008

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. Curr. Protoc. Immunol. 81:18.15.1-18.15.39. © 2008 by John Wiley & Sons, Inc.

Keywords:

  • sedimentation equilibrium;
  • sedimentation velocity;
  • chemical equilibria;
  • reversible interactions;
  • multi-protein complex;
  • analytical ultracentrifugation;
  • size-distribution;
  • Gilbert-Jenkins theory;
  • Lamm equation;
  • Bayesian analysis