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Accepted (AM)Open Access (License Unspecified), Open
Journal article
Determination of Protein Complex Stoichiometry Through Multisignal Sedimentation Velocity Experiments
Analytical biochemistry, v 407(1), pp 89-103
01 Dec 2010
PMID: 20667444
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Determination of the stoichiometry of macromolecular assemblies is fundamental to an understanding of how they function. Many different biophysical methodologies may be used to determine stoichiometry. In the past, both sedimentation equilibrium and sedimentation velocity analytical ultracentrifugation have been employed to determine component stoichiometries. Recently, a method of globally analyzing multisignal sedimentation velocity data was introduced by Schuck and colleagues. This global analysis removes some of the experimental inconveniences and inaccuracies that could occur in the previously used strategies. This method uses spectral differences between the macromolecular components to decompose the well-known
c
(
s
) distribution into component distributions
c
k
(
s
); i.e. each component
k
has its own
c
k
(
s
) distribution. Integration of these distributions allows for the calculation of the populations of each component in cosedimenting complexes, yielding their stoichiometry. In our laboratories, we have used this method extensively to determine the component stoichiometries of several protein-protein complexes involved in cytoskeletal remodeling, sugar metabolism, and host-pathogen interactions. The overall method is described in detail in this work, as well experimental examples and caveats.
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Details
- Title
- Determination of Protein Complex Stoichiometry Through Multisignal Sedimentation Velocity Experiments
- Creators
- Shae B. Padrick - The University of Texas Southwestern Medical CenterRanjit K. Deka - The University of Texas Southwestern Medical CenterJacinta L. Chuang - The University of Texas Southwestern Medical CenterR. Max Wynn - The University of Texas Southwestern Medical CenterDavid T. Chuang - The University of Texas Southwestern Medical CenterMichael V. Norgard - The University of Texas Southwestern Medical CenterMichael K. Rosen - Howard Hughes Medical InstituteChad A. Brautigam - The University of Texas Southwestern Medical Center
- Publication Details
- Analytical biochemistry, v 407(1), pp 89-103
- Publisher
- Elsevier
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Biochemistry and Molecular Biology
- Web of Science ID
- WOS:000282500500011
- Scopus ID
- 2-s2.0-77956884570
- Other Identifier
- 991020836366304721
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- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Biochemical Research Methods
- Biochemistry & Molecular Biology
- Chemistry, Analytical