A protocol for high throughput methods for the expression and purification of inner membrane proteins

McLuskey, K., Gabrielsen, M. , Kroner, F., Black, I., Cogdell, R.J. and Isaacs, N.W. (2008) A protocol for high throughput methods for the expression and purification of inner membrane proteins. Molecular Membrane Biology, 25(8), pp. 599-608. (doi: 10.1080/09687680802510289)

Abstract

The production of diffraction quality crystals for the structural determination of inner membrane proteins relies on obtaining large amounts of stable protein. Achieving this, by finding the correct parameters to successfully express and purify these proteins is often time-consuming and frustrating. The methods described here examine the most important parameters, in both expression and purification, quickly and simply. They take into account methods previously used in successful structural determinations of inner membrane proteins and collect and analyse data for use in further experiments and to investigate overall trends. These methods make use of histidine-tagged membrane proteins with a green fluorescent protein fusion but could be adapted easily for other proteins

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Black, Mrs Isobel and Gabrielsen, Dr Mads and McLuskey, Dr Karen and Cogdell, Professor Richard and Isaacs, Professor Neil
• Authors: McLuskey, K., Gabrielsen, M., Kroner, F., Black, I., Cogdell, R.J., and Isaacs, N.W.
• College/School: College of Medical Veterinary and Life Sciences > School of Molecular Biosciences; College of Science and Engineering > School of Chemistry
• Journal Name: Molecular Membrane Biology
• ISSN: 0968-7688