Purification, crystallization and X-ray crystallographic analysis of nicotinamidase Pnc1 from Kluyveromyces lactis

Pnc1 converts nicotinamide to nicotinic acid to generate NAD + through the Preiss-Handler pathway that is one of the NAD + -salvage pathway. By reducing levels of nicotinamide, an inhibitor of the NAD + -dependent histone deacetylase Sir2, yeast Pnc1 contributes gene silencing. In this study, to understand the structural features and molecular mechanism of nicotinamidase Pnc1, we overexpressed, purified, and crystallized the N-terminally His 6 -tagged Pnc1 protein from Kluyveromyces lactis and obtained X-ray diffraction data at a resolution of 2.2 Å. The crystals of the K. lactis Pnc1 (KlPnc1) belonged to space group P2 1 2 1 2 1 with unit cell parameters a=38.5, b=77.3, c=83.3, and α=β=γ= 90º. There is one molecule in the asymmetric unit.