Efficient production of membrane-integrated and detergent-soluble G protein-coupled receptors in Escherichia coli
Efficient production of membrane-integrated and detergent-soluble G protein-coupled receptors in Escherichia coli
复制标题
DOI：
10.1110/ps.035980.108
复制
发表时间：
2008-10-01
期刊：
影响因子：
8
通讯作者：
Georgiou, George
中科院分区：
文献类型：
--
作者：
Link, A. James;Skretas, Georgios;Georgiou, George
G protein-coupled receptors (GPCRs) are notoriously difficult to express, particularly in microbial systems. Using GPCR fusions with the green fluorescent protein (GFP), we conducted studies to identify bacterial host effector genes that result in a general and significant enhancement in the amount of membrane-integrated human GPCRs that can be produced in Escherichia coli. We show that coexpression of the membrane-bound AAA+ protease FtsH greatly enhances the expression yield of four different class I GPCRs, irrespective of the presence of GFP. Using this new expression system, we produced 0.5 and 2 mg/L of detergent-solubilized and purified full-length central cannabinoid receptor (CB1) and bradykinin receptor 2 (BR2) in shake flask cultures, respectively, two proteins that had previously eluded expression in microbial systems.
