Endogenous TOM20 Proximity Labeling: A Swiss-Knife for the Study of Mitochondrial Proteins in Human Cells
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-translat...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-05-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/11/9604 |
| _version_ | 1797597283278127104 |
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| author | Sébastien Meurant Lorris Mauclet Marc Dieu Thierry Arnould Sven Eyckerman Patricia Renard |
| author_facet | Sébastien Meurant Lorris Mauclet Marc Dieu Thierry Arnould Sven Eyckerman Patricia Renard |
| author_sort | Sébastien Meurant |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-11T03:04:21Z |
| format | Article |
| id | doaj.art-168c206f98d8442cb4dbe54d688bf915 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T03:04:21Z |
| publishDate | 2023-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-168c206f98d8442cb4dbe54d688bf9152023-11-18T08:01:21ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-05-012411960410.3390/ijms24119604Endogenous TOM20 Proximity Labeling: A Swiss-Knife for the Study of Mitochondrial Proteins in Human CellsSébastien Meurant0Lorris Mauclet1Marc Dieu2Thierry Arnould3Sven Eyckerman4Patricia Renard5URBC, Namur Research Institute for Life Sciences (Narilis), University of Namur (UNamur), 5000 Namur, BelgiumURBC, Namur Research Institute for Life Sciences (Narilis), University of Namur (UNamur), 5000 Namur, BelgiumMass Spectrometry Platform (MaSUN), Namur Research Institute for Life Sciences (Narilis), University of Namur (UNamur), 5000 Namur, BelgiumURBC, Namur Research Institute for Life Sciences (Narilis), University of Namur (UNamur), 5000 Namur, BelgiumVIB-UGent Center for Medical Biotechnology, VIB, 9000 Ghent, BelgiumURBC, Namur Research Institute for Life Sciences (Narilis), University of Namur (UNamur), 5000 Namur, BelgiumBiotin-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.https://www.mdpi.com/1422-0067/24/11/9604mitochondriaco-translational importBioIDprotein identificationmass spectrometry |
| spellingShingle | Sébastien Meurant Lorris Mauclet Marc Dieu Thierry Arnould Sven Eyckerman Patricia Renard Endogenous TOM20 Proximity Labeling: A Swiss-Knife for the Study of Mitochondrial Proteins in Human Cells International Journal of Molecular Sciences mitochondria co-translational import BioID protein identification mass spectrometry |
| title | Endogenous TOM20 Proximity Labeling: A Swiss-Knife for the Study of Mitochondrial Proteins in Human Cells |
| title_full | Endogenous TOM20 Proximity Labeling: A Swiss-Knife for the Study of Mitochondrial Proteins in Human Cells |
| title_fullStr | Endogenous TOM20 Proximity Labeling: A Swiss-Knife for the Study of Mitochondrial Proteins in Human Cells |
| title_full_unstemmed | Endogenous TOM20 Proximity Labeling: A Swiss-Knife for the Study of Mitochondrial Proteins in Human Cells |
| title_short | Endogenous TOM20 Proximity Labeling: A Swiss-Knife for the Study of Mitochondrial Proteins in Human Cells |
| title_sort | endogenous tom20 proximity labeling a swiss knife for the study of mitochondrial proteins in human cells |
| topic | mitochondria co-translational import BioID protein identification mass spectrometry |
| url | https://www.mdpi.com/1422-0067/24/11/9604 |
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