Measuring protein-protein interactions by equilibrium sedimentation.
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.
Balbo
Andrea
A
National Institutes of Health, Bethesda, Maryland.
Brown
Patrick H
PH
National Institutes of Health, Bethesda, Maryland.
Braswell
Emory H
EH
University of Connecticut, Storrs, Connecticut.
Schuck
Peter
P
University of Connecticut, Storrs, Connecticut.
D000818:Animals; D002623:Chemistry Techniques, Analytical / Q000295:instrumentation / Q000379:methods; D006801:Humans; D046911:Macromolecular Substances / Q000302:isolation & purification / Q000378:metabolism; D008962:Models, Theoretical; D011485:Protein Binding; D012107:Research Design; D012984:Software; D013223:Statistics as Topic / Q000379:methods; D013816:Thermodynamics; D014461:Ultracentrifugation / Q000295:instrumentation / Q000379:methods
D046911:Macromolecular Substances