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

DOI: 10.3109/09687688.2011.628954 DOI Help
PMID: 22034843 PMID Help

Authors: Mads Gabrielsen (University of Glasgow) , Frank Kroner (University of Glasgow) , Isobel Black , Neil Isaacs , Andrew J. Roe , Karen McLuskey (University of Glasgow)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Molecular Membrane Biology , VOL 28 (7-8) , PAGES 445 - 453

State: Published (Approved)
Published: October 2011

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 Escherichia coli. 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.

Journal Keywords: High-Throughput; Inner Transmembrane Protein; Detergents; Crystallization

Subject Areas: Biology and Bio-materials


Instruments: I03-Macromolecular Crystallography

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