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:Walker, Professor Daniel and Smith, Dr Brian and Byron, Professor Olwyn and Cogdell, Professor Richard and Grinter, Mr Rhys and Mosbahi, Dr Khedidja and Kelly, Dr Sharon and Roszak, Dr Aleksander
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