Keap1 cysteine 288 as a potential target for diallyl trisulfide-induced Nrf2 activation.
Keap1 cysteine 288 as a potential target for diallyl trisulfide-induced Nrf2 activation.
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DOI：
10.1371/journal.pone.0085984
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Surh YJ
中科院分区：
文献类型：
--
作者：
Kim S;Lee HG;Park SA;Kundu JK;Keum YS;Cha YN;Na HK;Surh YJ
二烯丙基硫化物、二烯丙基二硫化物和三硫化物（DATS）是大蒜油的主要挥发性成分。在本研究中，我们评估了它们诱导抗氧化酶表达的相对效力。在三种有机硫化合物中，DATS对人胃上皮细胞血红素加氧酶-1 （HO-1）和NAD(P)H：醌氧化还原酶-1 （NQO1）的诱导作用最强。此外，灌胃给药DATS可增加C57BL/6小鼠胃中HO-1和NQO1的表达。DATS处理可增加体外培养AGS细胞细胞核和小鼠胃内Nrf2的积累。在瞬时转染Nrf2- sirna的细胞或Nrf2缺失小鼠的胚胎成纤维细胞中，DATS诱导的HO-1和NQO1的表达被消除，这表明Nrf2是DATS细胞保护作用的关键介质。用n -乙酰半胱氨酸或二硫苏糖醇预处理AGS细胞可减弱dats诱导的Nrf2的核定位以及HO-1和NQO1的表达。Kelch-like ECH-associated protein-1 （Keap1）的半胱氨酸-151、-273和-288是Nrf2的胞浆抑制因子，被认为是氧化还原传感器，在Nrf2激活中发挥作用。为了确定DATS是否可以通过巯基修饰使Keap1失活，我们建立了表达野生型Keap1或三种不同突变结构的细胞系，其中Keap1的半胱氨酸-151、-273或-288被逆转录病毒基因转移取代。DATS不能激活Nrf2，也不能诱导HO-1和NQO1仅在Keap1-C288S突变细胞中表达。经DATS处理的重组Keap1的LC-ESI-MS/MS分析显示，含有Cys288的肽片段的分子质量为72.1 Da，相当于单烯丙基单硫化物的分子量。综上所述，这些发现表明DATS可能直接与Keap1的Cys288残基相互作用，这在一定程度上解释了DATS诱导Nrf2激活和上调防御基因表达的能力。
Diallyl sulfide, diallyl disulfide, and daillyl trisulfide (DATS) are major volatile components of garlic oil. In this study, we assessed their relative potency in inducing antioxidant enzyme expression. Among the three organosulfur compounds, DATS was found to be most potent in inducing heme oxygenase-1 (HO-1) and NAD(P)H:quinone oxidoreductase-1 (NQO1) in human gastric epithelial (AGS) cells. Furthermore, DATS administration by gavage increased the expression of HO-1 and NQO1 in C57BL/6 mouse stomach. Treatment with DATS increased the accumulation of nuclear factor-erythroid-2-related factor-2 (Nrf2) in the nucleus of cultured AGS cells and in mouse stomach in vivo. The DATS-induced expression of HO-1 and NQO1 was abrogated in the cells transiently transfected with Nrf2-siRNA or in the embryonic fibroblasts from Nrf2-null mice, indicating that Nrf2 is a key mediator of the cytoprotective effects of DATS. Pretreatment of AGS cells with N-acetylcysteine or dithiothreitol attenuated DATS-induced nuclear localization of Nrf2 and the expression of HO-1 and NQO1. Cysteine-151, -273 and -288 of Kelch-like ECH-associated protein-1 (Keap1), a cytosolic repressor of Nrf2, have been considered to act as a redox sensor and play a role in Nrf2 activation. To determine whether DATS could inactivate Keap1 through thiol modification, we established cell lines constitutively expressing wild type-Keap1 or three different mutant constructs in which cysteine-151, -273, or -288 of Keap1 was replaced with serine by retroviral gene transfer. DATS failed to activate Nrf2, and to induce expression of HO-1 and NQO1 only in Keap1-C288S mutant cells. LC-ESI-MS/MS analysis of recombinant Keap1 treated with DATS revealed that the peptide fragment containing Cys288 gained a molecular mass of 72.1 Da equivalent to the molecular weight of mono-allyl mono-sulfide. Taken together, these findings suggest that DATS may directly interact with the Cys288 residue of Keap1, which partly accounts for its ability to induce Nrf2 activation and upregulate defensive gene expression.
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影响因子：
5.3
作者：
Kobayashi, Makoto;Li, Li;Yamamoto, Masayuki
通讯作者：
Yamamoto, Masayuki
影响因子：
7.4
作者：
Munday, R;Munday, JS;Munday, CM
通讯作者：
Munday, CM
DOI：
10.1073/pnas.0914036107
发表时间：
2010-02-16
影响因子：
11.1
作者：
Ogura, Toshihiko;Tong, Kit I.;Yamamoto, Masayuki
通讯作者：
Yamamoto, Masayuki
影响因子：
254.7
作者：
Parkin, DM;Bray, F;Pisani, P
通讯作者：
Pisani, P
影响因子：
4.8
作者：
Alam, J;Stewart, D;Cook, JL
通讯作者：
Cook, JL