Open AccessProtein expression, crystallization and preliminary X‐ray crystallographic analysis of the isolated Shigella flexneri VapC toxin
Author(s) -
Xu Kehan,
Dedic Emil,
CobCantal Patricia,
Dienemann Christian,
Bøggild Andreas,
Winther Kristoffer S.,
Gerdes Kenn,
Brodersen Ditlev E.
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113014012
Subject(s) - shigella flexneri , crystallography , shigella , escherichia coli , crystallization , monoclinic crystal system , chemistry , biology , crystal structure , biochemistry , gene , organic chemistry
Upon release from the stable complex formed with its antitoxin VapB, the toxin VapC (MvpT) of the Gram‐negative pathogen Shigella flexneri is capable of globally down‐regulating translation by specifically cleaving initiator tRNA fMet in the anticodon region. Recombinant Shigella flexneri VapC D7A harbouring an active‐site mutation was overexpressed in Escherichia coli , purified to homogeneity and crystallized by the vapour‐diffusion technique. A preliminary X‐ray crystallographic analysis shows that the crystals diffracted to at least 1.9 Å resolution at a synchrotron X‐ray source and belonged to the trigonal space group in the hexagonal setting, H 3, with unit‐cell parameters a = b = 120.1, c = 52.5 Å, α = β = 90, γ = 120°. The Matthews coefficient is 2.46 Å 3 Da −1 , suggesting two molecules per asymmetric unit and corresponding to a solvent content of 50.0%.