Preliminary crystallographic analysis of sugar cane phosphoribosylpyrophosphate synthase

Abstract

Phosphoribosylpyrophosphate synthases (PRS; EC 2.7.6.1) are enzymes that are of central importance in several metabolic pathways in all cells. The sugar cane PRS enzyme contains 328 amino acids with a molecular weight of 36.6 kDa and represents the ®rst plant PRS to be crystallized, as well as the ®rst phosphateindependent PRS to be studied in molecular detail. Sugar cane PRS was overexpressed in Escherichia coli, puri®ed and crystallized using the hangingdrop vapour-diffusion method. Using X-ray diffraction experiments it was determined that the crystals belong to the orthorhombic system, with space group P2 1 2 1 2 and unit-cell parameters a = 213.2, b = 152.6, c = 149.3 A Ê . The crystals diffract to a maximum resolution of 3.3 A Ê and a complete data set to 3.5 A Ê resolution was collected and analysed.