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A novel yeast expression system for the overproduction of quality-controlled membrane proteins

Griffith, Douglas and Delipala, Christina and Leadsham, Jane E. and Jarvis, Simon M. and Oesterhelt, Dieter (2003) A novel yeast expression system for the overproduction of quality-controlled membrane proteins. F E B S Letters, 553 (1-2). pp. 45-50. ISSN 0014-5793

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Official URL: http://dx.doi.org/10.1016/S0014-5793(03)00952-9

Abstract

Saturation of the cell’s protein folding capacity and accumulation of inactive incompletely folded protein often accompanying the overexpression of membrane proteins (MPs) presents an obstacle to their efficient purification in a functional form for structural studies. We present a novel strategy for optimization of functional MP expression in Saccharomyces cerevisiae. This approach exploits the unfolded protein response (UPR) pathway, a stress signaling mechanism that senses the accumulation of unfolded proteins in the endoplasmic reticulum. We demonstrate that a high level of UPR induction upon expression of a MP reflects impaired functional expression of that protein. Tuning the expression level of the protein so as to avoid or minimize UPR induction results in its increased functional expression. UPR status can therefore serve as a proxy variable for the extent of impaired expression of a MP that may even be applicable in the absence of knowledge of the protein’s biological function.

Item Type:Article
Uncontrolled Keywords:Membrane protein, Protein overexpression, Protein folding, Unfolded protein response, Saccharomyces cerevisiae
Research Community:University of Westminster > Life Sciences, School of
ID Code:292
Deposited On:01 Dec 2005
Last Modified:07 Apr 2008 14:23

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