The C terminus of DJ-1 determines its homodimerization, MGO detoxification activity and suppression of ferroptosis
The C terminus of DJ-1 determines its homodimerization, MGO detoxification activity and suppression of ferroptosis
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DJ-1 的 C 末端决定其同源二聚化、MGO 解毒活性和铁死亡抑制
DOI：
10.1038/s41401-020-00531-1
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发表时间：
2020-10-06
影响因子：
8.2
通讯作者：
Cao, Ji
中科院分区：
文献类型：
--
作者：
Jiang, Li;Chen, Xiao-bing;Cao, Ji
DJ-1是一种与癌症和常染色体早发性帕金森病相关的多功能蛋白。DJ-1除了具有抗氧化应激活性外，最近的研究表明其具有去糖基化酶活性和抗铁凋亡作用。已经表明DJ-1形成同源二聚体，这决定了其抗氧化应激活性。在这项研究中，我们调查了DJ-1的二聚体结构和它的新报道的活动之间的关系。在具有Flag-taged和Myc-taged DJ-1过表达的HEK 293 T细胞中，我们进行了缺失突变和点突变，缩小了C末端最关键的基序。我们发现DJ-1的C端最后三个氨基酸的缺失突变（DJ-1 ΔC3）破坏了其同源二聚体，其中疏水性L187残基对DJ-1同源二聚体非常重要。此外，与野生型DJ-1（DJ-1 WT）相比，DJ-1 ΔC3突变和点突变L187 E几乎完全丧失了甲基乙二醛（MGO）的解毒和去糖基化能力。我们还发现，在DJ-1−/−小鼠胚胎成纤维细胞中，erastin触发的铁凋亡的抑制作用被ΔC3和L187 E消除，但被V51 C部分减弱。因此，我们的研究结果表明，DJ-1的C末端是至关重要的同源二聚化，去糖化活性，并抑制铁凋亡。
DJ-1 is a multifunctional protein associated with cancers and autosomal early-onset Parkinson disease. Besides the well-documented antioxidative stress activity, recent studies show that DJ-1 has deglycation enzymatic activity and anti-ferroptosis effect. It has been shown that DJ-1 forms the homodimerization, which dictates its antioxidative stress activity. In this study, we investigated the relationship between the dimeric structure of DJ-1 and its newly reported activities. In HEK293T cells with Flag-tagged and Myc-tagged DJ-1 overexpression, we performed deletion mutations and point mutations, narrowed down the most critical motif at the C terminus. We found that the deletion mutation of the last three amino acids at the C terminus of DJ-1 (DJ-1 ΔC3) disrupted its homodimerization with the hydrophobic L187 residue being of great importance for DJ-1 homodimerization. In addition, the ability in methylglyoxal (MGO) detoxification and deglycation was almost abolished in the mutation of DJ-1 ΔC3 and point mutant L187E compared with wild-type DJ-1 (DJ-1 WT). We also showed the suppression of erastin-triggered ferroptosis in DJ-1−/−mouse embryonic fibroblast cells was abolished by ΔC3 and L187E, but partially diminished by V51C. Thus, our results demonstrate that the C terminus of DJ-1 is crucial for its homodimerization, deglycation activity, and suppression of ferroptosis.