Expression, purification, crystallization and preliminary X-ray diffraction studies of glyceraldehyde-3-phosphate dehydrogenase 1 from methicillin-resistant Staphylococcus aureus (MRSA252).

Glyceraldehyde-3-phosphate dehydrogenase 1 from methicillin-resistant Staphylococcus aureus (MRSA252) was cloned in pQE30 vector, overexpressed in Escherichia coli M15(pREP4) cells and purified to homogeneity. The protein was crystallized using the hanging-drop vapour-diffusion method. The crystals belonged to space group P2(1), with unit-cell parameters a = 65.23, b = 95.58, c = 87.91 A, beta = 106.5 degrees . X-ray diffraction data were collected and processed to a maximum resolution of 2.0 A. The presence of one tetramer in the asymmetric unit gave a Matthews coefficient (V(M)) of 1.78 A(3) Da(-1) and a solvent content of 31%. The structure was solved by molecular replacement and structure refinement is now in progress.