Biochemical characterization of small heat shock protein HspB8 (Hsp22)-Bag3 interaction

Anton Shemetov, Nikolai B. Gusev

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Interaction of human Bag3 with small heat shock proteins HspB6, HspB8 and its K141E mutant was analyzed by different biochemical methods. The data of size-exclusion chromatography indicate that the wild type HspB8 forms tight complexes with Bag3. K141E mutant of HspB8 and especially HspB6 weaker interact with Bag3. The data of chemical crosslinking and analytical ultracentrifugation indicate that in vitro the stoichiometry of complexes formed by HspB8 and Bag3 is variable and is dependent on concentration of protein partners. Interaction of Bag3 and HspB8 is accompanied by increase of thermal stability measured by intrinsic tryptophan fluorescence and increased resistance to limited chymotrypsinolysis. The data of size-exclusion chromatography, analytical ultracentrifugation and limited proteolysis indicate that Bag3 belongs to the group of intrinsically disordered proteins. It is supposed that having unordered structure Bag3 might weakly interact with different small heat shock proteins which recognize unfolded proteins and this interaction is especially strong with intrinsically disordered HspB8. The complexes formed by Bag3 and HspB8 might have variable stoichiometry and can participate in different processes including clearing of the cell from improperly folded proteins.

Original languageEnglish (US)
Pages (from-to)1-9
Number of pages9
JournalArchives of Biochemistry and Biophysics
Volume513
Issue number1
DOIs
StatePublished - Sep 1 2011
Externally publishedYes

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Small Heat-Shock Proteins
Ultracentrifugation
Gel Chromatography
Size exclusion chromatography
Intrinsically Disordered Proteins
Stoichiometry
Protein Unfolding
Tryptophan
Proteolysis
Proteins
Hot Temperature
Fluorescence
Crosslinking
Thermodynamic stability
In Vitro Techniques

Keywords

  • Bag3
  • HspB6 (Hsp20)
  • HspB8 (Hsp22)
  • Phosphorylation
  • Protein-protein interaction
  • Small heat shock proteins

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

Biochemical characterization of small heat shock protein HspB8 (Hsp22)-Bag3 interaction. / Shemetov, Anton; Gusev, Nikolai B.

In: Archives of Biochemistry and Biophysics, Vol. 513, No. 1, 01.09.2011, p. 1-9.

Research output: Contribution to journalArticle

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