Expression, purification, crystallization and preliminary X-ray diffraction studies of phosphoglycerate kinase from methicillin-resistant Staphylococcus aureus MRSA252.

ABSTRACT: Phosphoglycerate kinase (PGK) from methicillin-resistant Staphylococcus aureus MRSA252 has been cloned in pQE30 expression vector, overexpressed in Escherichia coli SG13009 (pREP4) cells and purified to homogeneity. The protein was crystallized from 0.15?M CaCl(2), 0.1?M HEPES-NaOH pH 6.8, 20%(w/v) polyethylene glycol 2000 at 298?K by the hanging-drop vapour-diffusion method. The crystals belonged to space group P2(1), with unit-cell parameters a = 45.14, b = 74.75, c = 58.67?Å, ? = 95.72°. X-ray diffraction data have been collected and processed to a maximum resolution of 2.3?Å. The presence of one molecule in the asymmetric unit gives a Matthews coefficient (V(M)) of 2.26?Å(3)?Da(-1) with a solvent content of 46%. The structure has been solved by molecular replacement and structure refinement is now in progress.

SUBMITTER: Roychowdhury A 

PROVIDER: S-EPMC3107138 | biostudies-literature | 2011 Jun

REPOSITORIES: biostudies-literature