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For the Record

Efficient Production of Membrane-Integrated and Detergent-Soluble G Protein-Coupled Receptors in Escherichia coli

University of Texas at Austin

(RECEIVED April 24, 2008; ACCEPTED June 20, 2008)

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 of the amount of membrane-integrated human GPCRs that can be produced in Escherichia coli. We show that co-expression 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.

Keywords: Membrane Proteins; Expression systems; Escherichia coli; FtsH; G protein-coupled receptor; bradykinin receptor; cannabinoid receptor

¹ E-mail: gg{at}che.utexas.edu

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