Department of Medical, Oral and Biotechnological Sciences , University G. D'Annunzio Chieti , Chieti-Pescara, Via dei Vestini 31 , 66100 Chieti , Italy.
Proteomics and Metabonomics Unit , IRCCS Fondazione Santa Lucia , Via del Fosso di Fiorano 64 , 00143 Rome , Italy.
J Proteome Res. 2018 Dec 7;17(12):4307-4314. doi: 10.1021/acs.jproteome.8b00422. Epub 2018 Oct 5.
Mitochondria are undeniably the cell powerhouse, directly affecting cell survival and fate. Growing evidence suggest that mitochondrial protein repertoire affects metabolic activity and plays an important role in determining cell proliferation/differentiation or quiescence shift. Consequently, the bioenergetic status of a cell is associated with the quality and abundance of the mitochondrial populations and proteomes. Mitochondrial morphology changes in the development of different cellular functions associated with metabolic switches. It is therefore reasonable to speculate that different cell lines do contain different mitochondrial-associated proteins, and the investigation of these pools may well represent a source for mining missing proteins (MPs). A very effective approach to increase the number of IDs through mass spectrometry consists of reducing the complexity of the biological samples by fractionation. The present study aims at investigating the mitochondrial proteome of five phenotypically different cell lines, possibly expressing some of the MPs, through an enrichment-fractionation approach at the organelle and protein level. We demonstrate a substantial increase in the proteome coverage, which, in turn, increases the likelihood of detecting low abundant proteins, often falling in the category of MPs, and resulting, for the present study, in the identification of METTL12, FAM163A, and RGS13. All MS data have been deposited to the MassIVE data repository ( https://massive.ucsd.edu ) with the data set identifier MSV000082409 and PXD010446.
线粒体无疑是细胞的动力源，直接影响细胞的存活和命运。越来越多的证据表明，线粒体蛋白谱影响代谢活性，并在决定细胞增殖/分化或静止状态转变方面起着重要作用。因此，细胞的生物能量状态与线粒体群体和蛋白质组的质量和丰度有关。线粒体形态的变化与代谢转换相关的不同细胞功能的发展有关。因此，可以合理地推测，不同的细胞系确实含有不同的与线粒体相关的蛋白质，而对这些蛋白质池的研究很可能是挖掘缺失蛋白质（MPs）的一个来源。通过质谱法增加 ID 数量的一种非常有效的方法是通过分级分离来降低生物样品的复杂性。本研究旨在通过细胞器和蛋白质水平的富集分级分离方法，研究五种表型不同的细胞系的线粒体蛋白质组，这些细胞系可能表达一些 MPs。我们证明了蛋白质组覆盖范围的显著增加，这反过来又增加了检测低丰度蛋白质的可能性，这些蛋白质通常属于 MPs 类，因此，在本研究中，鉴定到了 METTL12、FAM163A 和 RGS13。所有 MS 数据都已被存入 MassIVE 数据库（https://massive.ucsd.edu），数据集标识符为 MSV000082409 和 PXD010446。
