The PINK1-Parkin mitophagy signalling pathway is not functional in peripheral blood mononuclear cells.
The PINK1-Parkin mitophagy signalling pathway is not functional in peripheral blood mononuclear cells.
复制标题
DOI：
10.1371/journal.pone.0259903
复制
发表时间：
2021
期刊：
影响因子：
3.7
通讯作者：
Taanman JW
中科院分区：
文献类型：
--
作者：
Bradshaw AV;Campbell P;Schapira AHV;Morris HR;Taanman JW
PINK 1和PRKN基因突变是早发性家族性帕金森病最常见的原因。这些基因分别编码PINK 1和Parkin蛋白，它们通过线粒体自噬参与功能失调的线粒体的降解。PINK 1-Parkin介导的线粒体自噬的早期步骤是线粒体融合蛋白MFN 1和MFN 2的泛素化。因此，患者样本中MFN 1和MFN-2的泛素化可以作为生物标志物来确定PINK 1和PRKN突变的功能效应，并筛选特发性患者的潜在线粒体自噬缺陷。我们的目的是检测PINK 1-Parkin线粒体自噬机制在外周血单个核细胞（PBMC）中的表达，并评估这些细胞是否可以作为一个平台，通过分析MFN 1和-2泛素化来评估线粒体自噬。通过质子载体CCCP处理通过线粒体去极化诱导线粒体自噬，并通过蛋白质印迹法分析泛素化的MFN蛋白。此外，PINK 1和PRKN mRNA和蛋白质表达水平分别采用逆转录酶定量PCR和蛋白质印迹法进行表征。虽然CCCP处理导致原代成纤维细胞、SH-SY 5 Y神经母细胞瘤细胞和Jurkat白血病细胞中的MFN泛素化，但PBMC处理不诱导MFN泛素化。PRKN mRNA和蛋白在PBMC中以与在Jurkat和成纤维细胞中观察到的水平相当的水平容易地检测到。相反，在对照PBMC中，PINK 1蛋白检测不到，PINK 1 mRNA水平显著低。我们的研究结果表明PINK 1-Parkin线粒体自噬信号通路在PBMC中不起作用。因此，PBMC不是用于帕金森病患者线粒体自噬功能分析的合适生物样品。
Mutations in the PINK1 and PRKN genes are the most common cause of early-onset familial Parkinson disease. These genes code for the PINK1 and Parkin proteins, respectively, which are involved in the degradation of dysfunctional mitochondria through mitophagy. An early step in PINK1 –Parkin mediated mitophagy is the ubiquitination of the mitofusin proteins MFN1 and -2. The ubiquitination of MFN1 and -2 in patient samples may therefore serve as a biomarker to determine the functional effects of PINK1 and PRKN mutations, and to screen idiopathic patients for potential mitophagy defects. We aimed to characterise the expression of the PINK1 –Parkin mitophagy machinery in peripheral blood mononuclear cells (PBMCs) and assess if these cells could serve as a platform to evaluate mitophagy via analysis of MFN1 and -2 ubiquitination. Mitophagy was induced through mitochondrial depolarisation by treatment with the protonophore CCCP and ubiquitinated MFN proteins were analysed by western blotting. In addition, PINK1 and PRKN mRNA and protein expression levels were characterised with reverse transcriptase quantitative PCR and western blotting, respectively. Whilst CCCP treatment led to MFN ubiquitination in primary fibroblasts, SH-SY5Y neuroblastoma cells and Jurkat leukaemic cells, treatment of PBMCs did not induce ubiquitination of MFN. PRKN mRNA and protein was readily detectable in PBMCs at comparable levels to those observed in Jurkat and fibroblast cells. In contrast, PINK1 protein was undetectable and PINK1 mRNA levels were remarkably low in control PBMCs. Our findings suggest that the PINK1 –Parkin mitophagy signalling pathway is not functional in PBMCs. Therefore, PBMCs are not a suitable biosample for analysis of mitophagy function in Parkinson disease patients.
登录
查看更多内容
影响因子：
64.8
作者：
通讯作者：
--
DOI：
10.1083/jcb.201801044
发表时间：
2018-05-07
期刊：
The Journal of cell biology
影响因子：
--
作者：
Lee JJ;Sanchez-Martinez A;Martinez Zarate A;Benincá C;Mayor U;Clague MJ;Whitworth AJ
通讯作者：
Whitworth AJ
影响因子：
2.5
作者：
Sunada, Y;Saito, F;Shimizu, T
通讯作者：
Shimizu, T
DOI：
10.1083/jcb.201402104
发表时间：
2014-04-28
期刊：
The Journal of cell biology
影响因子：
--
作者：
Kane LA;Lazarou M;Fogel AI;Li Y;Yamano K;Sarraf SA;Banerjee S;Youle RJ
通讯作者：
Youle RJ
DOI：
10.1083/jcb.200809125
发表时间：
2008-12-01
期刊：
The Journal of cell biology
影响因子：
--
作者：
Narendra D;Tanaka A;Suen DF;Youle RJ
通讯作者：
Youle RJ