title: Protein expression, crystallization and preliminary X-ray crystallographic analysis of the isolated Shigella flexneri VapC toxin.
abstract: 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, H3, 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%.
authors.affiliation: Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 10c, DK-8000 Aarhus C, Denmark.
authors.forename: Kehan
authors.initials: K
authors.lastname: Xu
authors.forename: Emil
authors.initials: E
authors.lastname: Dedic
authors.forename: Patricia
authors.initials: P
authors.lastname: Cob-Cantal
authors.forename: Christian
authors.initials: C
authors.lastname: Dienemann
authors.forename: Andreas
authors.initials: A
authors.lastname: Bøggild
authors.forename: Kristoffer S
authors.initials: KS
authors.lastname: Winther
authors.forename: Kenn
authors.initials: K
authors.lastname: Gerdes
authors.forename: Ditlev E
authors.initials: DE
authors.lastname: Brodersen
mesh_terms: D020134:Catalytic Domain; D003460:Crystallization; D018360:Crystallography, X-Ray; D004405:Dysentery, Bacillary / Q000235:genetics / Q000378:metabolism / Q000382:microbiology; D012358:RNA, Transfer, Met / Q000378:metabolism*; D022621:Shiga Toxin / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D012763:Shigella flexneri / Q000201:enzymology*; D017356:Synchrotrons
chemical_list: D012358:RNA, Transfer, Met; D022621:Shiga Toxin
keywords: PIN domain; Shigella flexneri; VapC; ribonucleases; tRNA; toxin–antitoxin