Expression, Purification, Crystallization and Preliminary X-ray Analysis of the Augmenter of Liver Regeneration (original) (raw)

Thiaminase type II (TenA) catalyzes the deamination of aminopyrimidines, including the cleavage of thiamine to 4-amino-5-hydroxymethyl-2-methylpyrimidine and 5-(2-hydroxyethyl)-4-methylthiazole in the metabolism of thiamine (vitamin B 1 ), in Staphylococcus aureus (Sa). SaTenA was crystallized by the vapour-diffusion method and the resulting crystal diffracted to 2.6 A resolution usng synchrotron radiation. The crystal is orthorhombic, belonging to space group P2 1 2 1 2 1 with unit-cell parameters a = 103.5, b = 104.1, c = 109.6 A ˚. With four molecules in the asymmetric unit, the Matthews coefficient is 2.85 A ˚3 Da À1 . Initial attempts to solve the structure by molecular-replacement techniques were successful.