URBC, Namur Research Institute for Life Sciences (Narilis), University of Namur (UNamur), 5000 Namur, Belgium.
Mass Spectrometry Platform (MaSUN), Namur Research Institute for Life Sciences (Narilis), University of Namur (UNamur), 5000 Namur, Belgium.
Int J Mol Sci. 2023 May 31;24(11):9604. doi: 10.3390/ijms24119604.
Biotin-based proximity labeling approaches, such as BioID, have demonstrated their use for the study of mitochondria proteomes in living cells. The use of genetically engineered BioID cell lines enables the detailed characterization of poorly characterized processes such as mitochondrial co-translational import. In this process, translation is coupled to the translocation of the mitochondrial proteins, alleviating the energy cost typically associated with the post-translational import relying on chaperone systems. However, the mechanisms are still unclear with only few actors identified but none that have been described in mammals yet. We thus profiled the TOM20 proxisome using BioID, assuming that some of the identified proteins could be molecular actors of the co-translational import in human cells. The obtained results showed a high enrichment of RNA binding proteins close to the TOM complex. However, for the few selected candidates, we could not demonstrate a role in the mitochondrial co-translational import process. Nonetheless, we were able to demonstrate additional uses of our BioID cell line. Indeed, the experimental approach used in this study is thus proposed for the identification of mitochondrial co-translational import effectors and for the monitoring of protein entry inside mitochondria with a potential application in the prediction of mitochondrial protein half-life.
基于生物素的邻近标记方法，如 BioID，已被证明可用于研究活细胞中的线粒体蛋白质组。使用基因工程化的 BioID 细胞系可以详细描述一些特征较差的过程，如线粒体共翻译导入。在这个过程中，翻译与线粒体蛋白的易位相偶联，减轻了通常与依赖伴侣系统的翻译后导入相关的能量成本。然而，目前只有少数几个已经确定的因子，但还没有在哺乳动物中描述过，因此机制仍不清楚。我们使用 BioID 对 TOM20 前导体进行了分析，假设鉴定出的一些蛋白质可能是人类细胞共翻译导入的分子因子。获得的结果显示，靠近 TOM 复合物的 RNA 结合蛋白高度富集。然而，对于少数选定的候选者，我们无法证明它们在线粒体共翻译导入过程中的作用。尽管如此，我们还是能够证明我们的 BioID 细胞系的其他用途。事实上，本研究中使用的实验方法可用于鉴定线粒体共翻译导入效应因子，并用于监测蛋白质进入线粒体的情况，这在预测线粒体蛋白半衰期方面具有潜在的应用。
