Open AccessOverexpression, purification, characterization and preliminary crystallographic study of phosphoglycolate phosphatase from Shigella flexneri 2a strain 301
Author(s) -
Liu Heli,
Zhou Huina,
Zhu Deyu,
Bi Ruchang
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309108039067
Subject(s) - shigella flexneri , strain (injury) , phosphatase , recombinant dna , chemistry , alkaline phosphatase , biochemistry , crystallography , escherichia coli , enzyme , biology , gene , anatomy
Phosphoglycolate phosphatase has a salvage function in the metabolism of the 2‐phosphoglycolate formed during bacterial DNA repair. In order to better understand its dimerization behaviour, the influence of metal ions on its activity and its catalytic mechanism at the molecular level, recombinant phosphoglycolate phosphatase from Shigella flexneri was overexpressed, purified, characterized and crystallized by the hanging‐drop vapour‐diffusion method at 291 K using polyethylene glycol 3500 as a precipitant and zinc acetate as an additive. The crystals belonged to space group R 3, with unit‐cell parameters a = 88.1, b = 88.1, c = 259.2 Å, corresponding to the presence of two molecules in the asymmetric unit. SeMet‐labelled protein was also prepared and crystallized for use in phase determination. Initial structure determination using the multiwavelength anomalous dispersion (MAD) method clearly revealed that SfPGPase bears an α‐helical cap domain that differs from that of a previously reported orthologue.