title: 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, β = 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.
authors.affiliation: Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur, India.
authors.forename: Amlan
authors.initials: A
authors.lastname: Roychowdhury
authors.forename: Somnath
authors.initials: S
authors.lastname: Mukherjee
authors.forename: Amit Kumar
authors.initials: AK
authors.lastname: Das
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D055624:Methicillin-Resistant Staphylococcus aureus / Q000201:enzymology*; D010735:Phosphoglycerate Kinase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D010735:Phosphoglycerate Kinase