Buneeva O A, Kopylov A T, Gnedenko O V, Medvedeva M V, Kapitsa I G, Ivanova E A, Ivanov A S, Medvedev A E
Institute of Biomedical Chemistry, Moscow, Russia.
Biological Faculty, Moscow State University, Moscow, Russia.
Biomed Khim. 2021 Jan;67(1):51-65. doi: 10.18097/PBMC20216701051.
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 play 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. (2020) Biochemistry (Moscow), Supplement Series B: Biomedical Chemistry, 14, 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 differentiation. Selected biosensor validation confirmed the interaction of Rpn13 subunit of proteasome with some proteins (glyceraldehyde-3-phosphate dehydrogenase, pyruvate kinase, histones H2A and H2B) revealed while proteomic profiling. The results obtained testify 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.
线粒体功能障碍和泛素-蛋白酶体系统（UPS）功能衰竭在帕金森病（PD）的发展中起着重要作用。位于调节性（19S）亚颗粒上的蛋白酶体亚基Rpn13在将待降解蛋白质递送至蛋白酶体的蛋白水解（20S）部分中发挥重要作用。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，《生物化学（莫斯科）》，增刊B：生物医学化学，2020年，第14卷，297 - 305页）。在本研究中，我们研究了神经毒素1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）和神经保护剂异吲哚酮对小鼠脑Rpn13结合蛋白的线粒体亚蛋白质组的影响。给动物注射MPTP（30 mg/kg）会导致典型的PD运动障碍，而异吲哚酮（100 mg/kg，在MPTP注射前30分钟）预处理可降低其严重程度。同时，注射MPTP、异吲哚酮或它们的组合（异吲哚酮 + MPTP）对Rpn13结合蛋白的总数和组成有显著影响。与对照组相比，注射MPTP减少了Rpn13结合蛋白的总数，而在MPTP注射前或不注射MPTP的情况下注射异吲哚酮会导致Rpn13结合蛋白数量显著增加，主要是参与蛋白质代谢调节、基因表达和分化的功能组蛋白。选定的生物传感器验证证实了蛋白酶体的Rpn13亚基与蛋白质组分析中发现的一些蛋白质（甘油醛-3-磷酸脱氢酶、丙酮酸激酶、组蛋白H2A和H2B）之间的相互作用。所得结果证明，在实验性MPTP诱导的帕金森病条件下，异吲哚酮的神经保护作用可能针对线粒体与UPS成分的相互作用。
