title: Cloning and characterization of NAD-dependent protein deacetylase (Rv1151c) from Mycobacterium tuberculosis.
abstract: Sir2 family proteins are highly conserved and catalyze a well-characterized NAD-dependent protein deacetylation reaction that regulates multiple cellular processes including aging, gene silencing, cellular differentiation, and metabolic pathways. Little is known about Sir2 family proteins in bacteria. The Sir2 homolog Rv1151c of Mycobacterium tuberculosis was cloned and over-expressed in Escherichia coli, and the protein then purified by Ni(2+)-affinity chromatography to homogeneity. The purified recombinant protein showed a typical NAD-dependent protein deacetylase activity that could be inhibited by nicotinamide and other known Sir2 inhibitors. The optimal temperature and pH for activity of Rv1151c are 25 degrees C and pH 9 +/- 1, respectively. Rv1151c is capable of deacetylating the acetyl-CoA synthetase from M. tuberculosis. However, unlike Sir2 family proteins identified from other bacteria, Rv1151c shows a substrate-independent NAD glycohydrolase activity in accordance with its auto-ADP ribosylation activity.
authors.affiliation: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, PR China.
authors.forename: Jing
authors.initials: J
authors.lastname: Gu
authors.forename: Jiao-Yu
authors.initials: JY
authors.lastname: Deng
authors.forename: Ru
authors.initials: R
authors.lastname: Li
authors.forename: Hongping
authors.initials: H
authors.lastname: Wei
authors.forename: Zhiping
authors.initials: Z
authors.lastname: Zhang
authors.forename: Yafeng
authors.initials: Y
authors.lastname: Zhou
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Xian-En
authors.initials: XE
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D004795:Enzyme Stability; D009169:Mycobacterium tuberculosis / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D037761:Sirtuins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D037761:Sirtuins