title: Escherichia coli as host for membrane protein structure determination: a global analysis.
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.
authors.affiliation: Laboratoire de Biologie Physico-Chimique des Protéines Membranaires, Institut de Biologie Physico-Chimique, CNRS, Univ Paris Diderot, Sorbonne Paris Cité, PSL research university, Paris, France.
authors.forename: Georges
authors.initials: G
authors.lastname: Hattab
authors.affiliation: Laboratoire de Biologie Physico-Chimique des Protéines Membranaires, Institut de Biologie Physico-Chimique, CNRS, Univ Paris Diderot, Sorbonne Paris Cité, PSL research university, Paris, France.
authors.forename: Dror E
authors.initials: DE
authors.lastname: Warschawski
authors.affiliation: Laboratoire de Biologie Physico-Chimique des Protéines Membranaires, Institut de Biologie Physico-Chimique, CNRS, Univ Paris Diderot, Sorbonne Paris Cité, PSL research university, Paris, France.
authors.forename: Karine
authors.initials: K
authors.lastname: Moncoq
authors.affiliation: Laboratoire de Biologie Physico-Chimique des Protéines Membranaires, Institut de Biologie Physico-Chimique, CNRS, Univ Paris Diderot, Sorbonne Paris Cité, PSL research university, Paris, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Miroux
mesh_terms: D001089:Arabinose / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics*; D015870:Gene Expression / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics
chemical_list: D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D014764:Viral Proteins; D001089:Arabinose; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases