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A novel yeast expression system for the overproduction of quality‐controlled membrane proteins
Author(s) -
Griffith Douglas A,
Delipala Christina,
Leadsham Jane,
Jarvis Simon M,
Oesterhelt Dieter
Publication year - 2003
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(03)00952-9
Subject(s) - unfolded protein response , endoplasmic reticulum , microbiology and biotechnology , saccharomyces cerevisiae , endoplasmic reticulum associated protein degradation , protein folding , yeast , membrane protein , function (biology) , biology , chemistry , biochemistry , membrane
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.