Posttranslational modification of Sirt6 activity by peroxynitrite.
Posttranslational modification of Sirt6 activity by peroxynitrite.
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DOI：
10.1016/j.freeradbiomed.2014.11.011
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发表时间：
2015-02
影响因子：
7.4
通讯作者：
Zhang, Wenbo
中科院分区：
文献类型：
--
作者：
Hu, Shugun;Liu, Hua;Ha, Yonju;Luo, Xuemei;Motamedi, Massoud;Gupta, Mahesh P.;Ma, Jian-Xing;Tilton, Ronald G.;Zhang, Wenbo
关键词：
哺乳动物sirtuin 6（Sirt 6）是一种位点特异性的组蛋白去乙酰化酶，调节染色质结构和许多基本的生物学过程。它抑制内皮细胞衰老和炎症，防止心脏肥大和心力衰竭的发展，调节葡萄糖代谢，并抑制肿瘤生长。调节Sirt 6酶功能的基本分子机制在很大程度上是未知的。在这里，我们假设Sirt 6功能可以通过翻译后修饰进行调节，重点是过氧亚硝酸盐的作用，这是疾病过程中产生的过量一氧化氮和超氧化物形成的主要活性氮物质之一。我们发现，孵育纯化的重组Sirt 6蛋白与3-morpholinosydnonimine（SIN-1，过氧亚硝酸盐供体，同时产生一氧化氮和超氧化物）增加Sirt 6酪氨酸硝化，降低其内在的催化活性。在SIN-1处理的Sirt 6（其在HEK 293细胞中过表达）中观察到类似的结果，并且当用SIN-1处理人视网膜微血管内皮细胞时，在内源性Sirt 6上观察到类似的结果。为了进一步研究Sirt 6硝化是否在病理条件下发生，我们使用内毒素诱导的视网膜炎症模型测定了视网膜中的Sirt 6硝化和活性。我们的数据表明，在该模型中，Sirt 6硝化增加，而其活性降低。通过质谱分析，我们鉴定了SIN-1处理后Sirt 6中的酪氨酸257被硝化。酪氨酸257突变为苯丙氨酸导致Sirt 6活性丧失，并消除SIN-1诱导的硝化和其活性降低。质谱分析还揭示了SIN-1处理后Sirt 6中甲硫氨酸和色氨酸的氧化。我们的研究结果表明，一种新的调节机制控制Sirt 6活性通过活性氮介导的翻译后修饰下的氧化和亚硝化应激。
The mammalian sirtuin 6 (Sirt6) is a site-specific histone deacetylase that regulates chromatin structure and many fundamental biological processes. It inhibits endothelial cell senescence and inflammation, prevents development of cardiac hypertrophy and heart failure, modulates glucose metabolism, and represses tumor growth. The basic molecular mechanisms underlying regulation of Sirt6 enzymatic function are largely unknown. Here we hypothesized that Sirt6 function can be regulated via posttranslational modification, focusing on the role of peroxynitrite, one of the major reactive nitrogen species formed by excessive nitric oxide and superoxide generated during disease processes. We found that incubation of purified recombinant Sirt6 protein with 3-morpholinosydnonimine (SIN-1, a peroxynitrite donor that generates nitric oxide and superoxide simultaneously) increased Sirt6 tyrosine nitration and decreased its intrinsic catalytic activity. Similar results were observed in SIN-1-treated Sirt6 which was overexpressed in HEK293 cells, and on endogenous Sirt6 when human retinal microvascular endothelial cells were treated with SIN-1. To further investigate whether Sirt6 nitration occurs under pathological conditions, we determined Sirt6 nitration and activity in retina using a model of endotoxin-induced retinal inflammation. Our data showed that Sirt6 nitration was increased while its activity was decreased in this model. With mass spectrometry, we identified that tyrosine 257 in Sirt6 was nitrated after SIN-1 treatment. Mutation of tyrosine 257 to phenylalanine caused loss of Sirt6 activity, and abolished SIN-1-induced nitration and decrease in its activity. Mass spectrometry analysis also revealed oxidation of methionine and tryptophan in Sirt6 after SIN-1 treatment. Our results demonstrate a novel regulatory mechanism controlling Sirt6 activity through reactive nitrogen species-mediated post-translational modification under oxidative and nitrosative stress.
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影响因子：
8.8
作者：
Lin Z;Yang H;Tan C;Li J;Liu Z;Quan Q;Kong S;Ye J;Gao B;Fang D
通讯作者：
Fang D
DOI：
10.1073/pnas.95.20.11584
发表时间：
1998-09-29
影响因子：
11.1
作者：
Kamisaki, Y;Wada, K;Murad, F
通讯作者：
Murad, F
DOI：
10.1016/s0006-291x(03)00814-3
发表时间：
2003-06-06
影响因子：
3.1
作者：
Ischiropoulos, H
通讯作者：
Ischiropoulos, H
影响因子：
--
作者：
El-Remessy AB;Tawfik HE;Matragoon S;Pillai B;Caldwell RB;Caldwell RW
通讯作者：
Caldwell RW
影响因子：
64.5
作者：
Kawahara TL;Michishita E;Adler AS;Damian M;Berber E;Lin M;McCord RA;Ongaigui KC;Boxer LD;Chang HY;Chua KF
通讯作者：
Chua KF