Multi-signal sedimentation velocity analysis with mass conservation for determining the stoichiometry of protein complexes.

ABSTRACT: Multi-signal sedimentation velocity analytical ultracentrifugation (MSSV) is a powerful tool for the determination of the number, stoichiometry, and hydrodynamic shape of reversible protein complexes in two- and three-component systems. In this method, the evolution of sedimentation profiles of macromolecular mixtures is recorded simultaneously using multiple absorbance and refractive index signals and globally transformed into both spectrally and diffusion-deconvoluted component sedimentation coefficient distributions. For reactions with complex lifetimes comparable to the time-scale of sedimentation, MSSV reveals the number and stoichiometry of co-existing complexes. For systems with short complex lifetimes, MSSV reveals the composition of the reaction boundary of the coupled reaction/mi

SUBMITTER: Brautigam CA 

PROVIDER: S-EPMC3656001 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature