Open AccessThe taming of small heat‐shock proteins: crystallization of the α‐crystallin domain from human Hsp27
Author(s) -
Baranova E. V.,
Beelen S.,
Gusev N. B.,
Strelkov S. V.
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109044571
Subject(s) - heat shock protein , hsp27 , crystallization , crystallography , proteolysis , protein subunit , materials science , biophysics , chemistry , biology , biochemistry , enzyme , hsp70 , organic chemistry , gene
Small heat‐shock proteins (sHsps) are ubiquitous molecular chaperones. sHsps function as homooligomers or heterooligomers that are prone to subunit exchange and structural plasticity. Here, a procedure for obtaining diffraction‐quality crystals of the α‐crystallin domain of human Hsp27 is presented. Initially, limited proteolysis was used to delineate the corresponding stable fragment (residues 90–171). This fragment could be crystallized, but examination of the crystals using X‐rays indicated partial disorder. The surface‐entropy reduction approach was applied to ameliorate the crystal quality. Consequently, a double mutant E125A/E126A of the 90–171 fragment yielded well ordered crystals that diffracted to 2.0 Å resolution.