Abstract—

Mitochondrial dysfunction and ubiquitin-proteasome system (UPS) failure contribute significantly to the development of Parkinson’s disease (PD). The proteasome subunit Rpn13 located on the regulatory (19S) subparticle plays an important role in the delivery of proteins, subjected to degradation, to the proteolytic (20S) part of proteasome. We have previously found several brain mitochondrial proteins specifically bound to Rpn13 (Buneeva et al., Biochemistry (Moscow), Supplement Series B: Biomedical Chemistry, (2020), vol. 14, pp. 297−305). In this study we have investigated the effect of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and the neuroprotector isatin on the mitochondrial subproteome of Rpn13-binding proteins of the mouse brain. Administration of MPTP (30 mg/kg) to animals caused movement disorders typical of PD, while pretreatment with isatin (100 mg/kg, 30 min before MPTP) reduced their severity. At the same time, the injection of MPTP, isatin, or their combination (isatin + MPTP) had a significant impact on the total number and the composition of Rpn13-binding proteins. The injection of MPTP decreased the total number of Rpn13-binding proteins in comparison with control, and the injection of isatin prior to MPTP or without MPTP caused an essential increase in the number of Rpn13-binding proteins, mainly of the functional group of proteins participating in the protein metabolism regulation, gene expression, and cell division and differentiation. Selected biosensor validation confirmed the interaction of the proteasome Rpn13 subunit with some proteins (glyceraldehyde-3-phosphate dehydrogenase, pyruvate kinase, histones H2A and H2B) recognized during proteomic profiling. The results obtained suggest that under the conditions of experimental MPTP-induced parkinsonism the neuroprotective effect of isatin may be aimed at the interaction of mitochondria with the components of UPS.

中文翻译：

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神经毒素 MPTP 和神经保护剂靛红诱导的小鼠脑 Rpn13 结合蛋白线粒体亚蛋白质组的变化

摘要——

线粒体功能障碍和泛素蛋白酶体系统 (UPS) 故障对帕金森病 (PD) 的发展有重大影响。位于调节 (19S) 亚颗粒上的蛋白酶体亚基 Rpn13 在将蛋白质递送至蛋白酶体的蛋白水解 (20S) 部分时发挥重要作用。我们之前已经发现了几种与 Rpn13 特异性结合的脑线粒体蛋白（Buneeva 等人，Biochemistry（莫斯科），Supplement Series B: Biomedical Chemistry, (2020), vol. 14，第 297-305 页）。在这项研究中，我们研究了神经毒素 1-甲基-4-苯基-1,2,3,6-四氢吡啶 (MPTP) 和神经保护剂靛红对小鼠大脑 Rpn13 结合蛋白的线粒体亚蛋白质组的影响。对动物施用 MPTP (30 mg/kg) 会导致典型的 PD 运动障碍，而用靛红 (100 mg/kg，MPTP 前 30 分钟) 预处理可降低其严重程度。同时，注射 MPTP、靛红或它们的组合（靛红 + MPTP）对 Rpn13 结合蛋白的总数和组成有显着影响。与对照相比，注射 MPTP 减少了 Rpn13 结合蛋白的总数，在 MPTP 或不使用 MPTP 之前注射靛红导致 Rpn13 结合蛋白的数量显着增加，主要是参与蛋白质代谢调控、基因表达和细胞分裂分化的蛋白质功能群。选定的生物传感器验证证实了蛋白酶体 Rpn13 亚基与蛋白质组学分析过程中识别的一些蛋白质（3-磷酸甘油醛脱氢酶、丙酮酸激酶、组蛋白 H2A 和 H2B）的相互作用。获得的结果表明，在实验性 MPTP 诱导的帕金森症条件下，靛红的神经保护作用可能是针对线粒体与 UPS 成分的相互作用。选定的生物传感器验证证实了蛋白酶体 Rpn13 亚基与蛋白质组学分析过程中识别的一些蛋白质（3-磷酸甘油醛脱氢酶、丙酮酸激酶、组蛋白 H2A 和 H2B）的相互作用。获得的结果表明，在实验性 MPTP 诱导的帕金森症条件下，靛红的神经保护作用可能是针对线粒体与 UPS 成分的相互作用。选定的生物传感器验证证实了蛋白酶体 Rpn13 亚基与蛋白质组学分析过程中识别的一些蛋白质（3-磷酸甘油醛脱氢酶、丙酮酸激酶、组蛋白 H2A 和 H2B）的相互作用。获得的结果表明，在实验性 MPTP 诱导的帕金森症条件下，靛红的神经保护作用可能是针对线粒体与 UPS 成分的相互作用。