Regulation of DJ-1 by Glutaredoxin 1 in Vivo: Implications for Parkinson's Disease.
Regulation of DJ-1 by Glutaredoxin 1 in Vivo: Implications for Parkinson's Disease.
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DOI：
10.1021/acs.biochem.5b01132
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发表时间：
2016-08-16
期刊：
影响因子：
2.9
通讯作者：
Mieyal JJ
中科院分区：
文献类型：
--
作者：
Johnson WM;Golczak M;Choe K;Curran PL;Miller OG;Yao C;Wang W;Lin J;Milkovic NM;Ray A;Ravindranath V;Zhu X;Wilson MA;Wilson-Delfosse AL;Chen SG;Mieyal JJ
帕金森病(PD)是世界上第二常见的神经退行性疾病，由黑质多巴胺能神经元变性引起。Park7(DJ-1)基因突变导致早发性常染色体隐性帕金森病，DJ-1的氧化修饰在体外调节DJ-1的保护活性。谷胱甘肽基化是一种普遍存在的蛋白质氧化还原修饰，由谷胱甘肽部分二硫键加成到反应性半胱氨酸-SH上而产生；特定蛋白质的谷胱甘肽基化与细胞活力的调节有关。谷氧还蛋白1(Grx1)是细胞内主要的谷胱甘肽合成酶，已被报道对线虫的多巴胺能神经元起到保护作用，然而Grx1在体内的许多下游功能靶点尚不清楚。以前，在模型神经元(SH-SY5Y和Neuro-2A系)的细胞培养中，DJ-1蛋白含量随着Grx1蛋白含量的减少而降低。在本研究中，我们旨在研究Grx1在体内对DJ-1的调节作用，并在体外表征其谷胱甘肽的作用。在这里，我们在Grx−/−小鼠身上提供了Grx1在体内调节DJ-1蛋白水平的证据。此外，在模型神经细胞(SH-SY5Y)中，我们观察到DJ-1蛋白含量在已知谷胱甘肽基化药物处理后降低；而在分离的DJ-1中，我们发现了两个不同的谷胱甘肽基化位点。最后，我们发现DJ-1在多巴胺能神经元中的过度表达可以部分补偿线虫体内帕金森病模型中Grx1同源基因的丢失。因此，我们的结果揭示了DJ-1的一种新的氧化还原修饰，并提示了一种新的体内DJ-1含量的调节机制。
Parkinson’s disease (PD) is the second most common neurodegenerative disease worldwide, caused by the degeneration of the dopaminergic neurons in the substantia nigra. Mutations in PARK7 (DJ-1) result in early onset autosomal recessive PD, and oxidative modification of DJ-1 has been reported to regulate the protective activity of DJ-1 in vitro. Glutathionylation is a prevalent redox modification of proteins resulting from the disulfide adduction of the glutathione moiety to a reactive cysteine-SH; and glutathionylation of specific proteins has been implicated in regulation of cell viability. Glutaredoxin 1 (Grx1) is the principal deglutathionylating enzyme within cells, and it has been reported to mediate protection of dopaminergic neurons in C. elegans, however many of the functional downstream targets of Grx1 in vivo remain unknown. Previously, DJ-1 protein content was shown to decrease concomitantly with diminution of Grx1 protein content in cell culture of model neurons (SH-SY5Y and Neuro-2A lines). In the current study we aimed to investigate the regulation of DJ-1 by Grx1 in vivo and characterize its glutathionylation in vitro. Here, with Grx−/− mice we provide evidence that Grx1 regulates protein levels of DJ-1 in vivo. Furthermore, with model neuronal cells (SH-SY5Y) we observed decreased DJ-1 protein content in response to treatment with known glutathionylating agents; and with isolated DJ-1 we identified two distinct sites of glutathionylation. Finally, we found that overexpression of DJ-1 in the dopaminergic neurons partly compensates for the loss of the Grx1 homolog in a C. elegans in vivo model of PD. Therefore; our results reveal a novel redox modification of DJ-1 and suggest a novel regulatory mechanism for DJ-1 content in vivo.