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Recombinant expression, purification, crystallization and preliminary X‐ray diffraction analysis of the C‐terminal DUF490 963–1138 domain of TamB from Escherichia coli
Author(s) -
Josts Inokentijs,
Grinter Rhys,
Kelly Sharon M.,
Mosbahi Khedidja,
Roszak Aleksander,
Cogdell Richard,
Smith Brian O.,
Byron Olwyn,
Walker Daniel
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x14017403
Subject(s) - escherichia coli , crystallography , crystallization , diffraction , bacteria , recombinant dna , chemistry , domain (mathematical analysis) , resolution (logic) , materials science , biology , biochemistry , physics , optics , gene , genetics , organic chemistry , mathematics , artificial intelligence , computer science , mathematical analysis
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 DUF490 963–1138 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 P 3 1 21, 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.