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

FEBS Letters

Jarvis, Simon M²  Leadsham, Jane²  Griffith, Douglas A¹  Oesterhelt, Dieter¹  Delipala, Christina¹ 

[1] Max-Planck-Institute for Biochemistry, Department of Membrane Biochemistry, Am Klopferspitz 18a, 82152 Martinsried, Germany;School of Biosciences, University of Westminster, 115 New Cavendish Street, London W1W 6UW, UK

关键词: Membrane protein;    Protein overexpression;    Protein folding;    Unfolded protein response;    Saccharomyces cerevisiae;   

DOI  :  10.1016/S0014-5793(03)00952-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown