Abstract
The Escherichia coli ClpA protein, like many AAA+ motor proteins, is allosterically regulated by nucleotide binding. We have combined analytical ultracentrifugation approaches with dynamic light scattering (DLS) to examine the self-association properties and the allosteric linkage of assembly to nucleotide binding. Here we present a protocol for the rapid and precise determination of the diffusion coefficient using DLS measurements in a model-independent fashion. When combined with analytical ultracentrifugation experiments, such an approach can yield a more complete understanding of the hydrodynamic and thermodynamic properties of the system.
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Acknowledgments
We thank Wilhelm Peters for alignment of the laser and pointing out many of the pitfalls discussed here. This work was supported by NSF grant MCB-0843746 to ALL and the University of Alabama Department of Chemistry Start-up Funds.
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Lucius, A.L., Veronese, P.K., Stafford, R.P. (2012). Dynamic Light Scattering to Study Allosteric Regulation. In: Fenton, A. (eds) Allostery. Methods in Molecular Biology, vol 796. Springer, New York, NY. https://doi.org/10.1007/978-1-61779-334-9_9
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DOI: https://doi.org/10.1007/978-1-61779-334-9_9
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