Preprint Article Version 2 Preserved in Portico This version is not peer-reviewed

Endogenous TOM20 Proximity Labelling: A Swiss-Knife for the Study of Mitochondrial Proteins in Human Cells

Version 1 : Received: 14 December 2022 / Approved: 16 December 2022 / Online: 16 December 2022 (10:29:37 CET)
Version 2 : Received: 20 April 2023 / Approved: 21 April 2023 / Online: 21 April 2023 (09:03:58 CEST)

A peer-reviewed article of this Preprint also exists.

Meurant, S.; Mauclet, L.; Dieu, M.; Arnould, T.; Eyckerman, S.; Renard, P. Endogenous TOM20 Proximity Labeling: A Swiss-Knife for the Study of Mitochondrial Proteins in Human Cells. Int. J. Mol. Sci. 2023, 24, 9604. Meurant, S.; Mauclet, L.; Dieu, M.; Arnould, T.; Eyckerman, S.; Renard, P. Endogenous TOM20 Proximity Labeling: A Swiss-Knife for the Study of Mitochondrial Proteins in Human Cells. Int. J. Mol. Sci. 2023, 24, 9604.

Abstract

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 mitochon-drial 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 prox-isome using BioID, assuming that some of identified proteins could be molecular actors of the co-translational import in human cells. The obtained results showed a high enrichment of RNA bind-ing 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 effec-tors and for the monitoring of protein entry inside mitochondria with a potential application in the prediction of mitochondrial protein half-life.

Keywords

Mitochondria; Co-translational import; BioID; Protein identification; Mass spectrometry

Subject

Biology and Life Sciences, Biology and Biotechnology

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