Dynamic light scattering to study allosteric regulation

Methods Mol Biol. 2012;796:175-86. doi: 10.1007/978-1-61779-334-9_9.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Allosteric Regulation / physiology
  • Endopeptidase Clp / chemistry*
  • Endopeptidase Clp / metabolism*
  • Escherichia coli Proteins / chemistry*
  • Escherichia coli Proteins / metabolism*
  • Light*
  • Protein Conformation
  • Scattering, Radiation
  • Thermodynamics
  • Ultracentrifugation / methods

Substances

  • Escherichia coli Proteins
  • ClpA protease, E coli
  • Endopeptidase Clp