Efficient production of membrane-integrated and detergent-soluble G protein-coupled receptors in Escherichia coli

A. James Link, Georgios Skretas, Eva Maria Strauch, Nandini S. Chari, George Georgiou

Research output: Contribution to journalArticle

47 Scopus citations

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 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.

Original languageEnglish (US)
Pages (from-to)1857-1863
Number of pages7
JournalProtein Science
Volume17
Issue number10
DOIs
StatePublished - Oct 2008

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Keywords

  • Bradykinin receptor
  • Cannabinoid receptor
  • Escherichia coli
  • FtsH
  • G protein-coupled receptor
  • Genetic engineering
  • Membrane protein

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