Expression of G-protein coupled receptors in Escherichia coli for structural studies

Biochemistry (Mosc). 2010 Jul;75(7):881-91. doi: 10.1134/s0006297910070102.

Abstract

To elaborate a high-performance system for expression of genes of G-protein coupled receptors (GPCR), methods of direct and hybrid expression of 17 GPCR genes in Escherichia coli and selection of strains and bacteria cultivation conditions were investigated. It was established that expression of most of the target GPCR fused with the N-terminal fragment of OmpF or Mistic using media for autoinduction provides high output (up to 50 mg/liter).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular
  • Escherichia coli / genetics*
  • Escherichia coli / metabolism
  • Gene Expression*
  • Humans
  • Multigene Family
  • Protein Conformation
  • Protein Structure, Tertiary
  • Receptors, G-Protein-Coupled / chemistry*
  • Receptors, G-Protein-Coupled / genetics*
  • Receptors, G-Protein-Coupled / metabolism
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • Receptors, G-Protein-Coupled
  • Recombinant Fusion Proteins