GPCR production in a novel yeast strain that makes cholesterol-like sterols

Kitson, S.M., Mullen, W. , Cogdell, R.J. , Bill, R.M. and Fraser, N.J. (2012) GPCR production in a novel yeast strain that makes cholesterol-like sterols. Methods, 55(4), pp. 287-292. (doi: 10.1016/j.ymeth.2011.09.023) (PMID:22001123)

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Publisher's URL: http://dx.doi.org/10.1016/j.ymeth.2011.09.023

Abstract

The activities of many mammalian membrane proteins including G-protein coupled receptors are cholesterol-dependent. Unlike higher eukaryotes, yeast do not make cholesterol. Rather they make a related molecule called ergosterol. As cholesterol and ergosterol are biologically non-equivalent, the potential of yeast as hosts for overproducing mammalian membrane proteins has never been fully realised. To address this problem, we are trying to engineer a novel strain of Saccharomyces cerevisiae in which the cholesterol biosynthetic pathway of mammalian cells has been fully reconstituted. Thus far, we have created a modified strain that makes cholesterol-like sterols which has an increased capacity to make G-protein coupled receptors compared to control yeast.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Fraser, Dr Niall and Kitson, Miss Susan and Mullen, Dr Bill and Cogdell, Professor Richard
Authors: Kitson, S.M., Mullen, W., Cogdell, R.J., Bill, R.M., and Fraser, N.J.
College/School:College of Medical Veterinary and Life Sciences > School of Infection & Immunity
College of Medical Veterinary and Life Sciences > School of Molecular Biosciences
Journal Name:Methods
ISSN:1046-2023
ISSN (Online):1095-9130)
Published Online:06 October 2011

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
443091Genetic manipulation of the sterol content of Pichia pastoris - a designer host for mammalian membrane protein productionNiall FraserBiotechnology and Biological Sciences Research Council (BBSRC)BB/E017061/1Institute of Cardiovascular and Medical Sciences