title: Recombinant expression, purification, crystallization and preliminary X-ray diffraction analysis of the C-terminal DUF490(963-1138) domain of TamB from Escherichia coli.
abstract: TamB is a recently described inner membrane protein that, together with its partner protein TamA, is required for the efficient secretion of a subset of autotransporter proteins in Gram-negative bacteria. In this study, the C-terminal DUF490963-1138 domain of TamB was overexpressed in Escherichia coli K-12, purified and crystallized using the sitting-drop vapour-diffusion method. The crystals belonged to the primitive trigonal space group P3121, with unit-cell parameters a = b = 57.34, c = 220.74 Å, and diffracted to 2.1 Å resolution. Preliminary secondary-structure and X-ray diffraction analyses are reported. Two molecules are predicted to be present in the asymmetric unit. Experimental phasing using selenomethionine-labelled protein will be undertaken in the future.
authors.affiliation: Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Scotland.
authors.forename: Inokentijs
authors.initials: I
authors.lastname: Josts
authors.affiliation: Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Scotland.
authors.forename: Rhys
authors.initials: R
authors.lastname: Grinter
authors.affiliation: Institute of Molecular Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Scotland.
authors.forename: Sharon M
authors.initials: SM
authors.lastname: Kelly
authors.affiliation: Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Scotland.
authors.forename: Khedidja
authors.initials: K
authors.lastname: Mosbahi
authors.affiliation: Institute of Molecular Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Scotland.
authors.forename: Aleksander
authors.initials: A
authors.lastname: Roszak
authors.affiliation: Institute of Molecular Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Scotland.
authors.forename: Richard
authors.initials: R
authors.lastname: Cogdell
authors.affiliation: Institute of Molecular Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Scotland.
authors.forename: Brian O
authors.initials: BO
authors.lastname: Smith
authors.affiliation: School of Life Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Scotland.
authors.forename: Olwyn
authors.initials: O
authors.lastname: Byron
authors.affiliation: Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Scotland.
authors.forename: Daniel
authors.initials: D
authors.lastname: Walker
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D017931:DNA Primers; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D017931:DNA Primers; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins
keywords: DUF490; Escherichia coli; TamB; autotransporter