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Efficient production of membrane‐integrated and detergent‐soluble G protein‐coupled receptors in Escherichia coli
Author(s) -
Link A. James,
Skretas Georgios,
Strauch EvaMaria,
Chari Nandini S.,
Georgiou George
Publication year - 2008
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.035980.108
Subject(s) - g protein coupled receptor , escherichia coli , green fluorescent protein , receptor , effector , membrane protein , biology , biochemistry , microbiology and biotechnology , chemistry , membrane , gene
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.