Cloning and characterization of NAD-dependent protein deacetylase (Rv1151c) from Mycobacterium tuberculosis

Jing Gu, Jiao Yu Deng, Ru Li, Hongping Wei, Zhiping Zhang, Yafeng Zhou, Ying Zhang, Xian En Zhang

Research output: Contribution to journalArticle

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 Ni2+-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°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.

Original languageEnglish (US)
Pages (from-to)743-748
Number of pages6
JournalBiochemistry (Moscow)
Volume74
Issue number7
DOIs
StatePublished - Jul 2009

Fingerprint

Cloning
Mycobacterium tuberculosis
NAD
Organism Cloning
Proteins
Bacteria
Acetate-CoA Ligase
NAD+ Nucleosidase
Affinity chromatography
Niacinamide
Escherichia coli Proteins
Gene Silencing
Metabolic Networks and Pathways
Affinity Chromatography
Recombinant Proteins
Adenosine Diphosphate
Genes
Aging of materials
Temperature
Substrates

Keywords

  • ADP-ribosyltransferase
  • Deacetylase
  • Mycobacterium tuberculosis
  • NAD glycohydrolase
  • Sir2

ASJC Scopus subject areas

  • Biochemistry
  • Medicine(all)

Cite this

Cloning and characterization of NAD-dependent protein deacetylase (Rv1151c) from Mycobacterium tuberculosis. / Gu, Jing; Deng, Jiao Yu; Li, Ru; Wei, Hongping; Zhang, Zhiping; Zhou, Yafeng; Zhang, Ying; Zhang, Xian En.

In: Biochemistry (Moscow), Vol. 74, No. 7, 07.2009, p. 743-748.

Research output: Contribution to journalArticle

Gu, Jing ; Deng, Jiao Yu ; Li, Ru ; Wei, Hongping ; Zhang, Zhiping ; Zhou, Yafeng ; Zhang, Ying ; Zhang, Xian En. / Cloning and characterization of NAD-dependent protein deacetylase (Rv1151c) from Mycobacterium tuberculosis. In: Biochemistry (Moscow). 2009 ; Vol. 74, No. 7. pp. 743-748.
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