Purification, crystallization and preliminary X-ray analysis of glucokinase from Streptomyces griseus in complex with glucose.

Glucokinase catalyzes the phosphorylation of glucose using ATP to yield glucose 6-phosphate. SgGlkA is a bacterial group III glucokinase from Streptomyces griseus that seems to play a regulatory role in carbon catabolite repression in this organism. SgGlkA was expressed in Escherichia coli, purified and crystallized using the sitting-drop vapour-diffusion method at 293†K. A crystal of SgGlkA in complex with glucose was obtained using a reservoir solution consisting of 0.9†M sodium/potassium tartrate, 0.2†M NaCl and 0.1†M imidazole pH 8.1 and diffracted X-rays to 1.84†à resolution. The crystal of SgGlkA in complex with glucose belonged to space group P6(2)22 or P6(4)22, with unit-cell parameters a = b = 109.19, c = 141.18†à . The crystal contained one molecule in the asymmetric unit.