title: Expression, purification, crystallization and preliminary X-ray diffraction studies of phosphoglycerate mutase from Staphylococcus aureus NCTC8325.
abstract: Phosphoglycerate mutase (PGM) is a key enzyme in carbohydrate metabolism. It takes part in both glycolysis and gluconeogenesis. PGM from pathogenic Staphylococcus aureus (NCTC8325) was cloned in pQE30 expression vector overexpressed in Escherichia coli M15 (pREP4) cells and purified to homogeneity. The protein was crystallized from two different conditions, (i) 0.1 M HEPES pH 7.5, 20%(w/v) polyethylene glycol 10,000 and (ii) 0.2 M NaCl, 0.1 M bis-tris pH 6.5, 25%(w/v) polyethylene glycol 3350, at 25°C by the sitting-drop vapour-diffusion method. Crystals grown at pH 7.5 diffracted to 2.5 Å resolution and belonged to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 77.0, b = 86.11, c = 94.07 Å. Crystals from the second condition at pH 6.5 diffracted to 2.00 Å resolution. These crystals belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 73.21, b = 81.75, c = 89.18 Å. X-ray diffraction data have been collected and processed to the maximum resolution to determine the structure of PGM. 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 721 302, India.
authors.forename: Amlan
authors.initials: A
authors.lastname: Roychowdhury
authors.affiliation: Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur 721 302, India.
authors.forename: Anirban
authors.initials: A
authors.lastname: Kundu
authors.affiliation: Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur 721 302, India.
authors.forename: Akanksha
authors.initials: A
authors.lastname: Gujar
authors.affiliation: Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur 721 302, India.
authors.forename: Madhuparna
authors.initials: M
authors.lastname: Bose
authors.affiliation: Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur 721 302, India.
authors.forename: Amit Kumar
authors.initials: AK
authors.lastname: Das
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D003460:Crystallization; D010736:Phosphoglycerate Mutase / Q000737:chemistry* / Q000302:isolation & purification*; D013211:Staphylococcus aureus / Q000201:enzymology*; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins; D010736:Phosphoglycerate Mutase
keywords: Staphylococcus aureus NCTC8325; phosphoglycerate mutase