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. 2015 Jul 10;5:12097.
doi: 10.1038/srep12097.

Escherichia Coli as Host for Membrane Protein Structure Determination: A Global Analysis

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Free PMC article

Escherichia Coli as Host for Membrane Protein Structure Determination: A Global Analysis

Georges Hattab et al. Sci Rep. .
Free PMC article

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.

Figures

Figure 1
Figure 1. Distribution of secondary structures in MP structures within the T7 expression system.
Membrane protein structures obtained from overexpression in the T7 system (102 see Table 1) were classified according to their secondary structure and topologies. For α-helical membrane proteins the number of transmembrane spans was represented as follow: monotopic (without transmembrane span), bitopic (1 transmembrane span) and integral membrane proteins (IMP, more than 1 transmembrane α-helices).
Figure 2
Figure 2. Expression protocol parameters in T7 and arabinose based expression systems.
Inducer concentrations and temperatures of growth were extracted using regular expression patterns in articles citing either Miroux and Walker, Studier and Moffatt or Guzman et al. for the recombinant expression of proteins in E. coli (see Materials and Methods). A. ITPG concentration in the T7 based expression system; B. temperature of growth in both T7 and arabinose based expression systems. Data are expressed as percentages of the total number of articles where an explicit value was found (See Table 2).

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