Polymerization of Oxidized DJ-1 via Noncovalent and Covalent Binding: Significance of Disulfide Bond Formation.
Polymerization of Oxidized DJ-1 via Noncovalent and Covalent Binding: Significance of Disulfide Bond Formation.
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氧化 DJ-1 通过非共价和共价键的聚合：二硫键形成的意义。
DOI：
10.1021/acsomega.9b00324
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发表时间：
2019
期刊：
影响因子：
4.1
通讯作者：
Saito Y
中科院分区：
文献类型：
--
作者：
Kobayashi M;Muramatsu K;Haruyama T;Uesugi H;Kikuchi A;Konno H;Noguchi N;Saito Y
DJ-1第106位的反应性半胱氨酸残基（Cys 106）在氧化应激下优先氧化，生成氧化DJ-1（oxDJ-1）。Cys 106氧化为亚磺酸改变了DJ-1的生物学作用，并增加了其细胞保护特性。类似的活化步骤是已知的过氧化物氧还蛋白（Prxs），其中反应性半胱氨酸的氧化亚磺酸诱导聚合的Prxs和改变其酶的特性从过氧化物酶的分子伴侣。在本研究中，oxDJ-1的制备和它的聚合和相关的氨基酸残基进行了研究。我们发现oxDJ-1形成了一个特征性的聚合物与二硫键和非共价和共价结合的二硫化物。谷胱甘肽的生理浓度解决了oxDJ-1的聚合物形式，并检测到其他两个半胱氨酸残基，如Cys 46和53的谷胱甘肽化。突变体分析表明，不仅Cys 106的必要性，而且Cys 46的聚合物形成。细胞实验表明，亲电醌处理诱导含有oxDJ-1的高分子量复合物。用原子力显微镜观察了oxDJ-1的动态聚合反应，发现oxDJ-1具有环状和堆叠结构。总的来说，这些结果清楚地证明了oxDJ-1与二硫键以及非共价和共价结合而不是二硫键形成特征性聚合物，这可能与oxDJ-1的生物学功能有关。
The reactive cysteine residue at position 106 (Cys106) of DJ-1 is preferentially oxidized under oxidative stress, generating oxidized DJ-1 (oxDJ-1). Oxidation of Cys106 to sulfinic acid changes the biologic action of DJ-1 and increases its cytoprotective properties. The similar activation step is known in peroxiredoxins (Prxs), in which oxidation of reactive Cys to sulfinic acid induces polymerization of Prxs and changes its enzyme characteristic from peroxidase to molecular chaperone. In the present study, oxDJ-1 was prepared and its polymerization and related amino acid residues were investigated. We found that oxDJ-1 formed a characteristic polymer with disulfide bonds and with noncovalent and covalent binding other than disulfide. The physiological concentration of glutathione resolved the polymer form of oxDJ-1, and glutathionylation of other two Cys residues, such as Cys 46 and 53, was detected. Mutant analysis indicated the necessity not only of Cys106 but also of Cys46 for the polymer formation. The cellular experiment demonstrated that the electrophilic quinone treatment induced a high-molecular-weight complex containing oxDJ-1. Dynamic polymerization of oxDJ-1 with a ring and a stacked structure was observed by an atomic force microscope. Collectively, these results clearly demonstrated the characteristic polymer formation of oxDJ-1 with a disulfide bond and noncovalent and covalent binding other than disulfide, which might be related to the biologic function of oxDJ-1.