Cloning, crystallization and preliminary X-ray analysis of a nucleotide-diphospho-sugar transferase spsA from Bacillus subtilis

Nucleotide-diphospho-sugar transferases represent, in terms of quantity, on e of the most important groups of enzymes on Earth, yet little is known abo ut their structure and mechanism. Such a transferase, the spsA gene product involved in the synthesis of the bacterial spore coat in Bacillus subtilis , has been cloned and overexpressed in an Escherichia coli expression syste m. Crystals have been grown, using PEG 8000 as a precipitant, in a form sui table for high-resolution X-ray analysis. They belong to space group C222(1 ), with unit-cell dimensions a = 42.4, b = 142.0, c = 81.4 Angstrom and wit h one molecule of spsA. in the asymmetric unit. The crystals diffract beyon d 1.5 Angstrom using synchrotron radiation.