Escherichia coli as host for membrane protein structure determination: a global analysis

Sci Rep. 2015 Jul 10:5:12097. doi: 10.1038/srep12097.

Abstract

The structural biology of membrane proteins (MP) is hampered by the difficulty in producing and purifying them. A comprehensive analysis of protein databases revealed that 213 unique membrane protein structures have been obtained after production of the target protein in E. coli. The primary expression system used was the one based on the T7 RNA polymerase, followed by the arabinose and T5 promoter based expression systems. The C41λ(DE3) and C43λ(DE3) bacterial mutant hosts have contributed to 28% of non E. coli membrane protein structures. A large scale analysis of expression protocols demonstrated a preference for a combination of bacterial host-vector together with a bimodal distribution of induction temperature and of inducer concentration. Altogether our analysis provides a set of rules for the optimal use of bacterial expression systems in membrane protein production.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabinose / genetics
  • Carrier Proteins / genetics
  • Cloning, Molecular / methods
  • DNA-Directed RNA Polymerases / genetics
  • Escherichia coli / genetics*
  • Escherichia coli Proteins / genetics*
  • Gene Expression / genetics
  • Genetic Vectors / genetics
  • Membrane Proteins / genetics*
  • Promoter Regions, Genetic / genetics
  • Viral Proteins / genetics

Substances

  • Carrier Proteins
  • Escherichia coli Proteins
  • Membrane Proteins
  • Viral Proteins
  • Arabinose
  • bacteriophage T7 RNA polymerase
  • DNA-Directed RNA Polymerases