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Mechanism of chaperone function in small heat shock proteins: Dissociation of the HSP27 oligomer is required for recognition and binding of destabilized T4 lysozyme
Journal of Biological Chemistry
  • Rangaiah Shashidharamurthy, Philadelphia College of Osteopathic Medicine
  • Hanane A. Koteiche
  • Jinhui Dong
  • Hassane S. Mchaoruab
Document Type
Article
Publication Date
1-1-2005
Disciplines
Abstract
Mammalian small heat shock proteins (sHSP) form polydisperse and dynamic oligomers that undergo equilibrium subunit exchange. Current models of their chaperone activity hypothesize that recognition and binding of protein non-native states involve changes in the oligomeric state. The equivalent thermodynamic representation is a set of three coupled equilibria that includes the sHSP oligomeric equilibrium, the substrate folding equilibrium, and the equilibrium binding between the sHSP and the substrate non-native states. To test this hypothesis and define the binding-competent oligomeric state of human Hsp27, we have perturbed the two former equilibria and quantitatively determined the consequences on binding. The substrate is a set of T4 lysozyme (T4L) mutants that bind under conditions that favor the folded state over the unfolded state by 10 2-10 4-fold. The concentration-dependent oligomer equilibrium of Hsp27 was perturbed by mutations that alter the relative stability of two major oligomeric states including phosphorylation-mimicking mutations that result in the dissociation to a small multimer over a wide range of concentrations. Correlation of binding isotherms with size exclusion chromatography analysis of the Hsp27 oligomer equilibrium demonstrates that the multimer is the binding-competent state. Binding occurs through two modes, each characterized by different affinity and number of binding sites, and results in T4L·Hsp27 complexes of different hydrodynamic properties. Mutants of the Hsp27 phosphorylation mimic that reverse the reduction in oligomer size also reduce the extent of T4L binding. Taken together, these results suggest a central role for the oligomeric equilibrium in regulating the chaperone activity of sHSP. The mutants identify sequence features important for modulating this equilibrium.
Comments

This article was published in Journal of Biological Chemistry, Volume 280, Issue 7, Pages 5281-5289.

The published version is available at http://dx.doi.org/10.1074/jbc.M407236200.

Copyright © 2005 ASBMB.

Citation Information
Rangaiah Shashidharamurthy, Hanane A. Koteiche, Jinhui Dong and Hassane S. Mchaoruab. "Mechanism of chaperone function in small heat shock proteins: Dissociation of the HSP27 oligomer is required for recognition and binding of destabilized T4 lysozyme" Journal of Biological Chemistry Vol. 280 Iss. 7 (2005) p. 5281 - 5289
Available at: http://works.bepress.com/rangaiah_shashidharamurthy/27/