Abstract

Article info Vol. 7  No. 1   pp.  24 ~ 27
Title Purification, crystallization and X-ray crystallographic analysis of nicotinamidase Pnc1 from Kluyveromyces lactis
Authors Shinae Kim and Jeong Ho Chang*
Institutions Department of Biology Education, Kyungpook National University, Daegu 41566, Republic of Korea *Correspondence: jhcbio@knu.ac.kr
Abstract 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 His6-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 P212121 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.