Measuring protein-protein interactions by equilibrium sedimentation

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

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.

MeSH terms

  • Animals
  • Chemistry Techniques, Analytical / instrumentation
  • Chemistry Techniques, Analytical / methods
  • Humans
  • Macromolecular Substances / isolation & purification
  • Macromolecular Substances / metabolism
  • Models, Theoretical
  • Protein Binding*
  • Research Design
  • Software
  • Statistics as Topic / methods
  • Thermodynamics
  • Ultracentrifugation* / instrumentation
  • Ultracentrifugation* / methods

Substances

  • Macromolecular Substances