The S-nitrosylation of parkin attenuated the ubiquitination of divalent metal transporter 1 in MPP(+)-treated SH-SY5Y cells.
The S-nitrosylation of parkin attenuated the ubiquitination of divalent metal transporter 1 in MPP(+)-treated SH-SY5Y cells.
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Parkin 的 S-亚硝基化减弱了 MPP 处理的 SH-SY5Y 细胞中二价金属转运蛋白 1 的泛素化
DOI：
10.1038/s41598-020-72630-2
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发表时间：
2020-09-23
影响因子：
4.6
通讯作者：
Jiang H
中科院分区：
文献类型：
--
作者：
Zhong Y;Li X;Du X;Bi M;Ma F;Xie J;Jiang H
由二价金属转运蛋白1 （DMT1）水平升高引起的异常铁积累有助于帕金森病（PD）的进行性神经变性。Parkin是DMT1泛素化的E3泛素连接酶。s -亚硝基化parkin （SNO-parkin）常见于PD。然而，s -亚硝基化对帕金森病中DMT1泛素化的E3泛素连接酶活性的影响在很大程度上是未知的。为了阐明s -亚硝基化parkin和DMT1在PD中的作用，我们用parkin转染SH-SY5Y细胞，并用s -亚硝基谷胱甘肽（GSNO）和1-甲基-4-苯基吡啶（MPP+）处理。结果显示，GSNO和MPP+处理后，一氧化氮（NO）和s -亚硝基化parkin的水平升高。一致地，观察到DMT1水平、铁摄取和细胞活力增加。有趣的是，抑制s -亚硝基化parkin降低了DMT1的水平。此外，parkin的s -亚硝基化显著抑制DMT1的泛素化。用单位点突变的parkin质粒（Cys241A, Cys260A, Cys323A）转染HEK293T细胞时，DMT1的泛素化也受到抑制。然而，共转染含有所有三种突变的质粒的细胞，GSNO处理不影响DMT1的泛素化。注射MPTP后2 h、4 h和24 h，大鼠黑质SNO-parkin和DMT1蛋白的表达逐渐升高。这些结果表明，parkin的s -亚硝基化抑制其E3泛素连接酶活性，从而抑制DMT1的泛素化，这有助于PD的铁积累和退行性过程。靶向s -亚硝基化可能为PD提供潜在的治疗策略。
Abnormal iron accumulation caused by elevated levels of divalent metal transporter 1 (DMT1) contributes to progressive neurodegeneration in Parkinson's disease (PD). Parkin is a E3 ubiquitin ligase for the ubiquitination of DMT1. S-nitrosylated parkin (SNO-parkin) is commonly observed in PD. However, the effects of S-nitrosylation on the E3 ubiquitin ligase activity of parkin for the ubiquitination of DMT1 in PD are largely unknown. To elucidate the role of S-nitrosylated parkin and DMT1 in PD, SH-SY5Y cells were transfected with parkin, being treated with S-nitrosoglutathione (GSNO) and 1-methyl-4-phenylpyridinium (MPP+). The results showed increased levels of oxidized nitric oxide (NO) and S-nitrosylated parkin after the treatment of GSNO and MPP+in parkin-transfected cells. Consistently, increased levels of DMT1, iron uptake and cell viability were observed. Interestingly, inhibition of S-nitrosylated parkin reduced the level of DMT1. Further, S-nitrosylation of parkin significantly inhibited the ubiquitination of DMT1. When HEK293T cells were transfected with plasmid of parkin with single site mutation (Cys241A, Cys260A, Cys323A), ubiquitination of DMT1 was also inhibited. However, the cells cotransfected with plasmids containing all three mutations, GSNO treatment did not affect the ubiquitination of DMT1. The expression of SNO-parkin and DMT1 protein in substantia nigra increased significantly gradually after 2 h, 4 h and 24 h with MPTP injection. These results indicate that the S-nitrosylation of parkin inhibits its E3 ubiquitin ligase activity for the ubiquitination of DMT1, which contributes to iron accumulation and degenerative process in PD. Targeted S-nitrosylation could provide a potential therapeutic strategy against PD.
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DOI：
10.1074/jbc.m115.703066
发表时间：
2016-05-06
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Liu K;Li F;Han H;Chen Y;Mao Z;Luo J;Zhao Y;Zheng B;Gu W;Zhao W
通讯作者：
Zhao W
影响因子：
3.3
作者：
Basu, Subhabrata;Mohan, Maradumane L.;Singh, Neena
通讯作者：
Singh, Neena
影响因子：
5.5
作者：
Chakraborty J;Basso V;Ziviani E
通讯作者：
Ziviani E
影响因子：
8.8
作者：
Lee Y;Stevens DA;Kang SU;Jiang H;Lee YI;Ko HS;Scarffe LA;Umanah GE;Kang H;Ham S;Kam TI;Allen K;Brahmachari S;Kim JW;Neifert S;Yun SP;Fiesel FC;Springer W;Dawson VL;Shin JH;Dawson TM
通讯作者：
Dawson TM
体外 ATP 敏感钾通道的激活增强 SK-N-SH 细胞中 DMT1 介导的铁摄取
影响因子：
4.6
作者：
Du X;Xu H;Shi L;Jiang Z;Song N;Jiang H;Xie J
通讯作者：
Xie J