Bacterial-based membrane protein production

Abstract

Escherichia coli is by far the most widely used bacterial host for the production of membrane proteins. Usually, different strains, culture conditions and production regimes are screened for to design the optimal production process. However, these E. coli-based screening approaches often do not result in satisfactory membrane protein production yields. Recently, it has been shown that (i) E. coli strains with strongly improved membrane protein production characteristics can be engineered or selected for, (ii) many membrane proteins can be efficiently produced in E. coli-based cell-free systems, (iii) bacteria other than E. coli can be used for the efficient production of membrane proteins, and, (iv) membrane protein variants that retain functionality but are produced at higher yields than the wild-type protein can be engineered or selected for. This article is part of a Special Issue entitled: Protein trafficking and secretion in bacteria. Guest Editors: Anastassios Economou and Ross Dalbey.

High-throughput recombinant protein expression in Escherichia coli : current status and future perspectives

“…Our understanding of how membrane proteins are translocated and folded in E. coli is highly limited, and it appears that the optimal strain for membrane protein production is protein-specific [144]. Currently, C41(DE3), C43(DE3) and Lemo21(DE3) remain the first-choice strains for membrane protein expression.…”

Stoichiometry determination of macromolecular membrane protein complexes

“…Both electron and X-ray crystallography depend on the successful crystallization of proteins and protein complexes. There have been many improvements in the production of membrane proteins for crystallization (Clark et al, 2011;Schlegel et al, 2014) and the formation and stabilization of crystals (Carpenter et al, 2008;Klara et al, 2016). However, as seen in crystal structures of components of the TtATPase, different subunits of type III secretion systems, as well as the nuclear pore complex (Lee et al, 2010;Worrall et al, 2010;Stuwe et al, 2015), mostly structures of soluble components or extramembrane domains of transmembrane proteins have been solved for large membrane-spanning complexes.…”

Les capteurs biomimétiques en recherche biomédicale

“…D'autres laboratoires ont développé des étiquettes peptidiques favorisant l'agrégation ou l'insertion des protéines dans les membranes, telles que Zera ou Mistic [8,9]. Mais ces solutions ne permettent pas actuellement d'entrevoir le début d'une normalisation de la production des protéines membranaires, car leur toxicité est multifactorielle (problème d'insertion dans les membranes, de Comme pour les protéines solubles, la production de protéines membranaires en Escherichia coli reste la plus largement utilisée [4]. Néanmoins, ce modèle n'est pas forcément le plus approprié pour la production de protéines membranaires d'origine eucaryote.…”