The E. coli sirtuin CobB shows no preference for enzymatic and nonenzymatic lysine acetylation substrate sites.
The E. coli sirtuin CobB shows no preference for enzymatic and nonenzymatic lysine acetylation substrate sites.
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DOI：
10.1002/mbo3.223
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发表时间：
2015-02
期刊：
影响因子：
3.4
通讯作者：
Wolfe, Alan J.
中科院分区：
文献类型：
--
作者：
AbouElfetouh, Alaa;Kuhn, Misty L.;Hu, Linda I.;Scholle, Michael D.;Sorensen, Dylan J.;Sahu, Alexandria K.;Becher, Doerte;Antelmann, Haike;Mrksich, Milan;Anderson, Wayne F.;Gibson, Bradford W.;Schilling, Birgit;Wolfe, Alan J.
关键词：
Nε-赖氨酸乙酰化是涉及多种细胞过程的数千种蛋白质的丰富翻译后修饰。在模型细菌大肠杆菌中，赖氨酸残基的 ε-氨基可通过乙酰辅酶 A (acCoA) 和赖氨酸乙酰转移酶催化乙酰化，或通过乙酰磷酸 (acP) 非酶促乙酰化。众所周知，催化 acCoA 依赖性 Nε-赖氨酸乙酰化可以通过脱乙酰酶逆转。在这里，我们提供了遗传、质谱、结构和免疫学证据，证明 CobB（NAD+ 依赖性脱乙酰酶的 Sirtuin 家族的一种脱乙酰酶）可以逆转乙酰化，无论乙酰供体或乙酰化机制如何。我们分析了 51 种蛋白质上的 69 个赖氨酸，我们之前检测到这些蛋白质在 cobB 突变体中比在其野生型亲本中具有稳健、可重复性并且显着更高的乙酰化程度。功能和通路富集分析支持了这样的假设：CobB 在多种且通常是必需的细胞过程（尤其是翻译）中调节蛋白质功能。结合质谱、生物信息学和蛋白质结构数据提供的证据表明，目标乙酰赖氨酸的可及性和三维微环境有助于确定 CobB 特异性。最后，我们提供证据表明 CobB 是大肠杆菌中主要的脱乙酰酶。
Nε-lysine acetylation is an abundant posttranslational modification of thousands of proteins involved in diverse cellular processes. In the model bacterium Escherichia coli, the ε-amino group of a lysine residue can be acetylated either catalytically by acetyl-coenzyme A (acCoA) and lysine acetyltransferases, or nonenzymatically by acetyl phosphate (acP). It is well known that catalytic acCoA-dependent Nε-lysine acetylation can be reversed by deacetylases. Here, we provide genetic, mass spectrometric, structural and immunological evidence that CobB, a deacetylase of the sirtuin family of NAD+-dependent deacetylases, can reverse acetylation regardless of acetyl donor or acetylation mechanism. We analyzed 69 lysines on 51 proteins that we had previously detected as robustly, reproducibly, and significantly more acetylated in a cobB mutant than in its wild-type parent. Functional and pathway enrichment analyses supported the hypothesis that CobB regulates protein function in diverse and often essential cellular processes, most notably translation. Combined mass spectrometry, bioinformatics, and protein structural data provided evidence that the accessibility and three-dimensional microenvironment of the target acetyllysine help determine CobB specificity. Finally, we provide evidence that CobB is the predominate deacetylase in E. coli.
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影响因子：
56.9
作者：
Choudhary, Chunaram;Kumar, Chanchal;Mann, Matthias
通讯作者：
Mann, Matthias
影响因子：
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作者：
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Escalante-Semerena, Jorge C.
DOI：
10.1107/s0907444904032147
发表时间：
2005-03-01
影响因子：
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作者：
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通讯作者：
Sukhareva, BS
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影响因子：
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作者：
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影响因子：
14.9
作者：
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通讯作者：
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