Open AccessExpression, refolding, crystallization and preliminary X‐ray analysis of Pseudomonas aeruginosa AlgE
Author(s) -
Whitney John C. C.,
Neculai A. Mirela,
Ohman Dennis E.,
Howell P. Lynne
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430910901094x
Subject(s) - crystallization , pseudomonas aeruginosa , chemistry , crystallography , diffraction , molecule , strain (injury) , x ray crystallography , chromatography , bacteria , biology , physics , organic chemistry , optics , genetics , anatomy
AlgE is an outer membrane protein present in alginate‐producing (mucoid) Pseudomonas aeruginosa . AlgE has been overexpressed in insoluble inclusion bodies, purified under denaturing conditions and refolded in a buffer containing decyl β‐ d ‐maltopyranoside. Purified refolded AlgE was detergent‐exchanged into n ‐octyl tetraoxyethylene and diffraction‐quality crystals were grown using the hanging‐drop vapor‐diffusion method. The crystals grew as small hexagons with unit‐cell parameters a = 98.8, b = 156.8, c = 90.4 Å, α = β = γ = 90.0°. The crystals exhibited the symmetry of space group C 222 and diffracted to a minimum d ‐spacing of 3.0 Å. On the basis of the Matthews coefficient ( V M = 3.28 Å 3 Da −1 ), one molecule is estimated to be present in the asymmetric unit.