title: Expression, purification, crystallization and preliminary crystallographic analysis of MxiH, a subunit of the Shigella flexneri type III secretion system needle.
abstract: A monodisperse truncation mutant of MxiH, the subunit of the needle from the Shigella flexneri type III secretion system (TTSS), has been overexpressed and purified. Crystals were grown of native and selenomethionine-labelled MxiH(CDelta5) and diffraction data were collected to 1.9 A resolution. The crystals belong to space group C2, with unit-cell parameters a = 183.4, b = 28.1, c = 27.8 A, beta = 96.5 degrees. An anomalous difference Patterson map calculated with the data from the SeMet-labelled crystals revealed a single peak on the Harker section v = 0. Inspection of a uranyl derivative also revealed one peak in the isomorphous difference Patterson map on the Harker section v = 0. Analysis of the self-rotation function indicates the presence of a twofold non-crystallographic symmetry axis approximately along a. The calculated Matthews coefficient is 1.9 A3 Da(-1) for two molecules per asymmetric unit, corresponding to a solvent content of 33%.
authors.affiliation: Department of Biochemistry, University of Oxford, England.
authors.forename: Janet E
authors.initials: JE
authors.lastname: Deane
authors.forename: Frank S
authors.initials: FS
authors.lastname: Cordes
authors.forename: Pietro
authors.initials: P
authors.lastname: Roversi
authors.forename: Steven
authors.initials: S
authors.lastname: Johnson
authors.forename: Roma
authors.initials: R
authors.lastname: Kenjale
authors.forename: William D
authors.initials: WD
authors.lastname: Picking
authors.forename: Wendy L
authors.initials: WL
authors.lastname: Picking
authors.forename: Susan M
authors.initials: SM
authors.lastname: Lea
authors.forename: Ariel
authors.initials: A
authors.lastname: Blocker
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D012763:Shigella flexneri / Q000737:chemistry* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; C413572:MxiH protein, Shigella; D011994:Recombinant Proteins