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Accepted (AM)Open Access (License Unspecified), Open
Journal article
Evaluating the stoichiometry of macromolecular complexes using multisignal sedimentation velocity
Methods (San Diego, Calif.), v 54(1)
01 May 2011
PMID: 21256217
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Gleaning information regarding the molecular physiology of macromolecular complexes requires knowledge of their component stoichiometries. In this work, a relatively new means of analyzing sedimentation velocity (SV) data from the analytical ultracentrifuge is examined in detail. The method depends on collecting concentration profile data simultaneously using multiple signals, like Rayleigh interferometry and UV spectrophotometry. If the cosedimenting components of a complex are spectrally distinguishable, continuous sedimentation-coefficient distributions specific for each component can be calculated to reveal the molar ratio of the complex's components. When combined with the hydrodynamic information available from the SV data, a stoichiometry can be derived. Herein, the spectral properties of sedimenting species are systematically explored to arrive at a predictive test for whether a set of macromolecules can be spectrally resolved in a multisignal SV (MSSV) experiment. Also, a graphical means of experimental design and criteria to judge the success of the spectral discrimination in MSSV are introduced. A detailed example of the analysis of MSSV experiments is offered, and the possibility of deriving equilibrium association constants from MSSV analyses is explored. Finally, successful implementations of MSSV are reviewed. (C) 2011 Elsevier Inc. All rights reserved.
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Details
- Title
- Evaluating the stoichiometry of macromolecular complexes using multisignal sedimentation velocity
- Creators
- Shae B. Padrick - The University of Texas Southwestern Medical CenterChad A. Brautigam - Applied BioPhysics (United States)
- Publication Details
- Methods (San Diego, Calif.), v 54(1)
- Publisher
- Elsevier
- Number of pages
- 17
- Grant note
- F32GM069179 / NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of General Medical Sciences (NIGMS) R01-GM56322; 1F32-GM06917902 / National Institutes of Health (NIH); United States Department of Health & Human Services; National Institutes of Health (NIH) - USA
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Biochemistry and Molecular Biology
- Web of Science ID
- WOS:000290649700005
- Scopus ID
- 2-s2.0-79955639129
- Other Identifier
- 991020836338404721
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InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Web of Science research areas
- Biochemical Research Methods
- Biochemistry & Molecular Biology