Recombinant expression, purification, crystallization and preliminary X-ray diffraction analysis of the C-terminal DUF490963–1138 domain of TamB from Escherichia coli

Josts, I., Grinter, R., Kelly, S. M. , Mosbahi, K., Roszak, A., Cogdell, R. , Smith, B. O. , Byron, O. and Walker, D. (2014) Recombinant expression, purification, crystallization and preliminary X-ray diffraction analysis of the C-terminal DUF490963–1138 domain of TamB from Escherichia coli. Acta Crystallographica. Section F: Structural Biology Communications, 70(9), pp. 1272-1275. (doi: 10.1107/S2053230X14017403)

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Publisher's URL: http://dx.doi.org/10.1107/S2053230X14017403

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.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Mosbahi, Dr Khedidja and Kelly, Dr Sharon and Roszak, Dr Aleksander and Walker, Professor Daniel and Smith, Dr Brian and Byron, Professor Olwyn and Cogdell, Professor Richard and Grinter, Mr Rhys
Authors: Josts, I., Grinter, R., Kelly, S. M., Mosbahi, K., Roszak, A., Cogdell, R., Smith, B. O., Byron, O., and Walker, D.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cardiovascular and Medical Sciences
College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
College of Science and Engineering > School of Chemistry
College of Medical Veterinary and Life Sciences > School of Life Sciences
College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Acta Crystallographica. Section F: Structural Biology Communications
Publisher:International Union of Crystallography
ISSN:1744-3091
ISSN (Online):2053-230X
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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
558531Wellcome Trust Studentship 2010-14Daniel WalkerWellcome Trust (WELLCOME)093597/Z/10/ZIII - BACTERIOLOGY
558532Wellcome Trust Studentship 2010-14Daniel WalkerWellcome Trust (WELLCOME)093597/Z/10/ZIII - BACTERIOLOGY