Open AccessExpression, purification, crystallization and preliminary X‐ray analysis of Pseudomonas aeruginosa PelD
Author(s) -
Marmont Lindsey S.,
Whitney John C.,
Robinson Howard,
Colvin Kelly M.,
Parsek Matthew R.,
Howell P. Lynne
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111052109
Subject(s) - pseudomonas aeruginosa , crystallization , microbiology and biotechnology , chemistry , materials science , crystallography , biology , bacteria , genetics , organic chemistry
The production of the PEL polysaccharide in Pseudomonas aeruginosa requires the binding of bis‐(3′,5′)‐cyclic dimeric guanosine monophosphate (c‐di‐GMP) to the cytoplasmic GGDEF domain of the inner membrane protein PelD. Here, the overexpression, purification and crystallization of a soluble construct of PelD that encompasses the GGDEF domain and a predicted GAF domain is reported. Diffraction‐quality crystals were grown using the hanging‐drop vapour‐diffusion method. The crystals grew as flat plates, with unit‐cell parameters a = 88.3, b = 114.0, c = 61.9 Å, α = β = γ = 90.0°. The PelD crystals exhibited the symmetry of space group P 2 1 2 1 2 and diffracted to a minimum d ‐spacing of 2.2 Å. On the basis of the Matthews coefficient ( V M = 2.29 Å 3 Da −1 ), it was estimated that two molecules are present in the asymmetric unit.