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

Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
Isabelle BroutinArnaud Ducruix

Abstract

OprM and OprN belong to the outer membrane factor family proteins. These approximately 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-beta-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 A, beta = 98.9 degrees and a = 151.3, b = 87.6, c = 356.5 A, beta = 98.1 degrees for OprM and OprN, respectively. Using the ESRF synchrotron-radiation source, OprM diffraction data extended to 3.4 A.

References

Sep 1, 1995·Antimicrobial Agents and Chemotherapy·X Z LiK Poole
Sep 22, 2001·Acta Crystallographica. Section D, Biological Crystallography·J Navaza
May 15, 2003·International Journal of Toxicology·Patrick F McDermottDavid G White
Sep 3, 2003·The Journal of Antimicrobial Chemotherapy·Yang LiTomofusa Tsuchiya
Mar 3, 2004·Acta Crystallographica. Section D, Biological Crystallography·Laurent C StoroniRandy J Read

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Citations

Apr 2, 2009·PloS One·Myriam ReffayWladimir Urbach
Jun 4, 2015·Langmuir : the ACS Journal of Surfaces and Colloids·Isabelle MottaFrederic Pincet
Aug 5, 2014·The Journal of Membrane Biology·Yann FerrandezPhilippe Minard
Oct 13, 2007·Proteins·Juliette MartinAnne-Claude Camproux
May 17, 2012·Electrophoresis·Yann FerrandezIsabelle Broutin
Sep 9, 2017·PloS One·Yvette Véronique Ntsogo EnguénéIsabelle Broutin

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