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

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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.

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Abbreviations

ACS:

acetyl-CoA synthetase

GDH:

glutamate dehydrogenase

HNA:

2-hydroxy-1-naphthaldehyde

LB medium:

Luria-Bertani medium

NA:

nicotinic acid

NAM:

nicotinamide

OAADPr:

O-acetyl-ADP-ribose

POA:

pyrazinoic acid

PZA:

pyrazinamide

Sir2:

silent information regulator 2

TB:

tuberculosis

ZMAL:

(S)-[5-acetylamino-1-(4-methyl-2-oxo-2H-chromen-7-ylcarbamoyl)-pentyl]-carbamic acid benzyl ester

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Authors and Affiliations

  1. State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, P. R. China

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

  2. Graduate School, Chinese Academy of Science, Beijing, 100039, China

    Jing Gu & Ru Li

  3. Department of Molecular Microbiology and Immunology, School of Hygiene and Public Health, Johns Hopkins University, Baltimore, MD, 21205, USA

    Ying Zhang

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  1. Jing Gu
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  2. Jiao-Yu Deng
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  3. Ru Li
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  4. Hongping Wei
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  5. Zhiping Zhang
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  6. Yafeng Zhou
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  7. Ying Zhang
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  8. Xian-En Zhang
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Correspondence to Xian-En Zhang.

Additional information

Published in Russian in Biokhimiya, 2009, Vol. 74, No. 7, pp. 916–922.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM08-400, February 8, 2009.

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Gu, J., Deng, JY., Li, R. et al. Cloning and characterization of NAD-dependent protein deacetylase (Rv1151c) from Mycobacterium tuberculosis . Biochemistry Moscow 74, 743–748 (2009). https://doi.org/10.1134/S0006297909070062

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  • DOI: https://doi.org/10.1134/S0006297909070062

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