Abstract
Analytical ultracentrifugation (AUC) is a very useful technique to characterize macromolecular interactions. In AUC, a centrifugal force of up to about 250,000 g is applied to a solution of macromolecules, and the progression of sedimentation over time is monitored using an optical detection system. Significant advances in both hardware and software over the past few decades have greatly improved the applicability of AUC for the study of protein–protein interactions. The purpose of this chapter is to provide experimental strategies for the analysis of protein–protein interactions using AUC, including the determination of the association constant of self-associations, binding stoichiometry, and equilibrium binding constant of heterogeneous protein–protein associations. An overview of the method and software packages available for AUC data analysis and optimal protocols for the characterization of protein–protein interactions will be described.
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Krayukhina, E., Uchiyama, S. (2016). Analytical Ultracentrifugation. In: Senda, T., Maenaka, K. (eds) Advanced Methods in Structural Biology. Springer Protocols Handbooks. Springer, Tokyo. https://doi.org/10.1007/978-4-431-56030-2_10
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DOI: https://doi.org/10.1007/978-4-431-56030-2_10
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