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Expression, purification, crystallization and preliminary X‐ray studies of the outer membrane efflux proteins OprM and OprN from Pseudomonas aeruginosa

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OprM and OprN belong to the outer membrane factor family proteins. These ∼52 kDa proteins are part of the tripartite efflux pumps found in Pseudomonas aeruginosa and are responsible in part for the antibiotic resistance observed in these bacteria. Both proteins have been expressed in Escherichia coli as His‐tag proteins and purified accordingly by affinity chromatography in the presence of n‐octyl‐β‐d‐glucopyranoside detergent. OprM and OprN were crystallized using PEG 20 000/ammonium citrate and ammonium sulfate as precipitating agents, respectively. Crystals belong to space group C2, with unit‐cell parameters a = 152.6, b = 87.9, c = 355.9 Å, β = 98.9° and a = 151.3, b = 87.6, c = 356.5 Å, β = 98.1° for OprM and OprN, respectively. Using the ESRF synchrotron‐radiation source, OprM diffraction data extended to 3.4 Å.
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Keywords: Efflux pumps; antibiotic resistance; outer membrane factor family

Document Type: Research Article

Affiliations: Laboratoire de Cristallographie et RMN Biologiques, UMR 8015 CNRS, Faculté de Pharmacie, Université René Descartes, 4 Avenue de l'Observatoire, 75270 Paris CEDEX 06, France

Publication date: March 1, 2005

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