Open AccessCloning, expression, purification, crystallization and preliminary X‐ray analysis of the 31 kDa Vibrio cholerae heat‐shock protein VcHsp31
Author(s) -
Das Samir,
Dey Sanjay,
Roy Trina,
Sen Udayaditya
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111032970
Subject(s) - vibrio cholerae , crystallization , cloning (programming) , heat shock protein , shock (circulatory) , chemistry , biology , microbiology and biotechnology , biochemistry , bacteria , genetics , gene , medicine , computer science , organic chemistry , programming language
The Gram‐negative bacterium Vibrio cholerae , which is responsible for the diarrhoeal disease cholera in humans, induces the expression of numerous heat‐shock genes. VcHsp31 is a 31 kDa putative heat‐shock protein that belongs to the DJ‐1/PfpI superfamily, functioning as both a chaperone and a protease. VcHsp31 has been cloned, overexpressed and purified by Ni 2+ –NTA affinity chromatography followed by gel filtration. Crystals of VcHsp31 were grown in the presence of PEG 6000 and MPD; they belonged to space group P 2 1 and diffracted to 1.9 Å resolution. Assuming the presence of six molecules in the asymmetric unit, the Matthews coefficient was estimated to be 1.97 Å 3 Da −1 , corresponding to a solvent content of 37.4%.