Efficient production of membrane‐integrated and detergent‐soluble G protein‐coupled receptors inEscherichia coli

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Abstract
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 inEscherichia 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.