Open AccessThe purification, crystallization and preliminary diffraction of a glycerophosphodiesterase from Enterobacter aerogenes
Author(s) -
Jackson Colin J.,
Carr Paul D.,
Kim HyeKyung,
Liu JianWei,
Ollis David L.
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106020021
Subject(s) - enterobacter aerogenes , crystallization , escherichia coli , enzyme , chemistry , crystallography , monomer , solvent , biochemistry , organic chemistry , gene , polymer
The metallo‐glycerophosphodiesterase from Enterobacter aerogenes (GpdQ) has been cloned, expressed in Escherichia coli and purified. Initial screening of crystallization conditions for this enzyme resulted in the identification of needles from one condition in a sodium malonate grid screen. Removal of the metals from the enzyme and subsequent optimization of these conditions led to crystals that diffracted to 2.9 Å and belonged to space group P 2 1 3, with unit‐cell parameter a = 164.1 Å. Self‐rotation function analysis and V M calculations indicated that the asymmetric unit contains two copies of the monomeric enzyme, corresponding to a solvent content of 79%. It is intended to determine the structure of this protein utilizing SAD phasing from transition metals or molecular replacement.