Journal
METHODS
Volume 55, Issue 4, Pages 287-292Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2011.09.023
Keywords
Yeast; Cholesterol; Membrane protein; GPCR; Overexpression; Genetic modification
Funding
- National Institute of General Medical Sciences [R21 GM078182-01]
- British Heart Foundation [CH/98001]
- BBSRC [BB/E008720/1, BB/E017061/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/E017061/1, BB/E008720/1] Funding Source: researchfish
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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. (C) 2011 Elsevier Inc. All rights reserved.
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