High-throughput identification of purification conditions leads to preliminary crystallization conditions for three inner membrane proteins

Gabrielsen, M., Kroner, F., Black, I., Isaacs, N.W., Roe, A.J. and McLuskey, K. (2011) High-throughput identification of purification conditions leads to preliminary crystallization conditions for three inner membrane proteins. Molecular Membrane Biology, 28(7-8), pp. 445-453. (doi:10.3109/09687688.2011.628954)

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Abstract

An important factor in the crystallization, and subsequent structural determination, of integral membrane proteins is the ability to produce a stable and monodisperse solution of the protein. Obtaining the correct purification detergent to achieve this can be laborious and is often serendipitous. In this study, high-throughput methods are used to analyze the suitability of eight different detergents on the stability of 12 inner transmembrane proteins from <i>Escherichia coli</i>. The best results obtained from the small-scale experiments were scaled up, the aggregation state of the proteins assessed, and all monodisperse protein solutions entered into crystallization trials. This resulted in preliminary crystallization hits for three inner membrane proteins: XylH, PgpB and YjdL and this study reports the methods, purification procedures and crystallization conditions used to achieve this.

Item Type:Articles
Keywords:High-throughput, inner transmembrane protein, detergents, crystallization
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:McLuskey, Dr Karen and Roe, Professor Andrew and Isaacs, Professor Neil and Black, Mrs Isobel and Gabrielsen, Dr Mads
Authors: Gabrielsen, M., Kroner, F., Black, I., Isaacs, N.W., Roe, A.J., and McLuskey, K.
Subjects:Q Science > QH Natural history > QH345 Biochemistry
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
College of Science and Engineering > School of Chemistry
Journal Name:Molecular Membrane Biology
Publisher:Informa Healthcare
ISSN:0968-7688

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
491441A biochemical and molecular analysis of the YhaO membrane protein in Escherichia coli O157:H7Andrew RoeBiotechnology and Biological Sciences Research Council (BBSRC)BB/G011389/1Infection Immunity and Inflammation Life Sciences