Open AccessPurification, crystallization and preliminary X‐ray diffraction analysis of Cif, a virulence factor secreted by Pseudomonas aeruginosa
Author(s) -
Bahl Christopher D.,
MacEachran Daniel P.,
O'Toole George A.,
Madden Dean R.
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109047599
Subject(s) - pseudomonas aeruginosa , virulence factor , cystic fibrosis transmembrane conductance regulator , crystallization , virulence , transmembrane protein , cystic fibrosis , chemistry , atp binding cassette transporter , microbiology and biotechnology , recombinant dna , biology , crystallography , biochemistry , bacteria , transporter , genetics , gene , receptor , organic chemistry
The opportunistic pathogen Pseudomonas aeruginosa secretes a protein that triggers the accelerated degradation of the cystic fibrosis transmembrane conductance regulator (CFTR) in airway epithelial cells. This protein, which is known as the CFTR inhibitory factor (Cif), acts as a virulence factor and may facilitate airway colonization by P. aeruginosa . Based on sequence similarity Cif appears to be an epoxide hydrolase (EH), but it lacks several of the conserved features found in the active sites of canonical members of the EH family. Here, the crystallization of purified recombinant Cif by vapor diffusion is reported. The crystals formed in space group C 2, with unit‐cell parameters a = 167.4, b = 83.6, c = 88.3 Å, β = 100.6°. The crystals diffracted to 2.39 Å resolution on a rotating‐anode source. Based on the calculated Matthews coefficient (2.2 Å 3 Da −1 ), it appears that the asymmetric unit contains four molecules.