Regions Outside the Alpha-Crystallin Domain of the Small Heat Shock Protein Hsp26 Are Required for Its Dimerization

Journal of Molecular Biology, 2010, doi:10.1016/j.jmb.2010.02.022, Volume 398, Issue 1, Pages 122-131 published on 18.02.2010
Journal of Molecular Biology, online article
Small heat shock proteins (sHsps) are a ubiquitous family of molecular chaperones. They form homo-oligomers, composed of mostly 24 subunits. The immunoglobulin-like α-crystallin domain, which is flanked by N- and C-terminal extensions, is the most conserved element in sHsps. It is assumed to be the dimeric building block from which the sHsp oligomers are assembled. Hsp26 from Saccharomyces cerevisiae is a well-characterized member of this family.With a view to study the structural stability and oligomerization properties of its alpha-crystallin domain, we produced a series of alpha-crystallin domain constructs. We show that a minimal α-crystallin domain can, against common belief, be monomeric and stably folded. Elongating either the N- or the C-terminus of this minimal αlpha-crystallin domain with the authentic extensions leads to the formation of dimeric species. In the case of N-terminal extensions, their population is dependent on the presence of the complete so-called Hsp26 “middle domain”. For the C-terminal extensions, the presence of the conserved IXI motif of sHsps is necessary and sufficient to induce dimerization, which can be inhibited by increasing ionic strength. Dimerization does not induce major changes in secondary structure of the Hsp26 αlpha-crystallin domain. A thermodynamic analysis of the monomeric and dimeric constructs revealed that dimers are not significantly stabilized against thermal and chemical denaturation in comparison to monomers, supporting our notion that dimerization is not a prerequisite for the formation of a well-folded Hsp26 αlpha-crystallin domain.

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