Cloning and characterization of NAD-dependent protein deacetylase (Rv1151c) from Mycobacterium tuberculosis
Cloning and characterization of NAD-dependent protein deacetylase (Rv1151c) from Mycobacterium tuberculosis
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DOI：
10.1134/s0006297909070062
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发表时间：
2009-07-01
影响因子：
2.8
通讯作者：
Zhang, Xian-En
中科院分区：
文献类型：
--
作者：
Gu, Jing;Deng, Jiao-Yu;Zhang, Xian-En
Sir2家族蛋白是高度保守的，并催化一种特征明确的nad依赖性蛋白去乙酰化反应，调节多种细胞过程，包括衰老、基因沉默、细胞分化和代谢途径。我们对细菌中的Sir2家族蛋白知之甚少。克隆结核分枝杆菌Sir2同源基因Rv1151c，并在大肠杆菌中过表达，用Ni2+亲和层析纯化该蛋白，使其均匀性。纯化后的重组蛋白具有典型的nad依赖性蛋白去乙酰化酶活性，可被烟酰胺和其他已知的Sir2抑制剂抑制。Rv1151c活性的最适温度为25℃，pH为9 +/- 1。Rv1151c能够使结核分枝杆菌的乙酰辅酶a合成酶去乙酰化。然而，与从其他细菌中鉴定出的Sir2家族蛋白不同，Rv1151c显示出与底物无关的NAD糖水解酶活性，与其自adp核糖基化活性一致。
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 25A 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.