Open AccessOverexpression, purification, crystallization and preliminary X‐ray analysis of CheY4 from Vibrio cholerae O395
Author(s) -
Biswas Maitree,
Khamrui Susmita,
Sen Udayaditya,
Dasgupta Jhimli
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111041972
Subject(s) - vibrio cholerae , motility , escherichia coli , chemotaxis , microbiology and biotechnology , biology , virulence factor , gene , virulence , chemistry , bacteria , biochemistry , genetics , receptor
Chemotaxis and motility greatly influence the infectivity of Vibrio cholerae , although the role of chemotaxis genes in V. cholerae pathogenesis is poorly understood. In contrast to the single copy of CheY found in Escherichia coli and Salmonella typhimurium , four CheYs (CheY1–CheY4) are present in V. cholerae . While insertional disruption of the cheY4 gene results in decreased motility, insertional duplication of this gene increases motility and causes enhanced expression of the two major virulence genes. Additionally, cheY3 / cheY4 influences the activation of the transcription factor NF‐κB, which triggers the generation of acute inflammatory responses. V. cholerae CheY4 was cloned, overexpressed and purified by Ni–NTA affinity chromatography followed by gel filtration. Crystals of CheY4 grown in space group C 2 diffracted to 1.67 Å resolution, with unit‐cell parameters a = 94.4, b = 31.9, c = 32.6 Å, β = 96.5°, whereas crystals grown in space group P 3 2 21 diffracted to 1.9 Å resolution, with unit‐cell parameters a = b = 56.104, c = 72.283 Å, γ = 120°.