Proteomic mapping of cytosol-facing outer mitochondrial and ER membranes in living human cells by proximity biotinylation
The cytosol-facing membranes of cellular organelles contain proteins that enable signal transduction, regulation of morphology and trafficking, protein import and export, and other specialized processes. Discovery of these proteins by traditional biochemical fractionation can be plagued with contami...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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eLife Sciences Publications, Ltd.
2017
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| Online Access: | http://hdl.handle.net/1721.1/109872 https://orcid.org/0000-0003-3972-2820 https://orcid.org/0000-0002-2687-3470 https://orcid.org/0000-0002-7203-4299 https://orcid.org/0000-0002-8277-5226 |
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| author | Udeshi, Namrata D Svinkina, Tanya Hung, Victoria Lam, Stephanie Shih-Min Guzman, Gaelen Donnelly Mootha, Vamsi Carr, Steven A Ting, Alice Y |
| author2 | Broad Institute of MIT and Harvard |
| author_facet | Broad Institute of MIT and Harvard Udeshi, Namrata D Svinkina, Tanya Hung, Victoria Lam, Stephanie Shih-Min Guzman, Gaelen Donnelly Mootha, Vamsi Carr, Steven A Ting, Alice Y |
| author_sort | Udeshi, Namrata D |
| collection | MIT |
| description | The cytosol-facing membranes of cellular organelles contain proteins that enable signal transduction, regulation of morphology and trafficking, protein import and export, and other specialized processes. Discovery of these proteins by traditional biochemical fractionation can be plagued with contaminants and loss of key components. Using peroxidase-mediated proximity biotinylation, we captured and identified endogenous proteins on the outer mitochondrial membrane (OMM) and endoplasmic reticulum membrane (ERM) of living human fibroblasts. The proteomes of 137 and 634 proteins, respectively, are highly specific and highlight 94 potentially novel mitochondrial or ER proteins. Dataset intersection identified protein candidates potentially localized to mitochondria-ER contact sites. We found that one candidate, the tail-anchored, PDZ-domain-containing OMM protein SYNJ2BP, dramatically increases mitochondrial contacts with rough ER when overexpressed. Immunoprecipitation-mass spectrometry identified ribosome-binding protein 1 (RRBP1) as SYNJ2BP’s ERM binding partner. Our results highlight the power of proximity biotinylation to yield insights into the molecular composition and function of intracellular membranes. |
| first_indexed | 2024-09-23T10:49:05Z |
| format | Article |
| id | mit-1721.1/109872 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T10:49:05Z |
| publishDate | 2017 |
| publisher | eLife Sciences Publications, Ltd. |
| record_format | dspace |
| spelling | mit-1721.1/1098722022-09-30T23:16:19Z Proteomic mapping of cytosol-facing outer mitochondrial and ER membranes in living human cells by proximity biotinylation Udeshi, Namrata D Svinkina, Tanya Hung, Victoria Lam, Stephanie Shih-Min Guzman, Gaelen Donnelly Mootha, Vamsi Carr, Steven A Ting, Alice Y Broad Institute of MIT and Harvard Massachusetts Institute of Technology. Department of Chemistry Hung, Victoria Lam, Stephanie Shih-Min Guzman, Gaelen Donnelly Mootha, Vamsi Carr, Steven A Ting, Alice Y The cytosol-facing membranes of cellular organelles contain proteins that enable signal transduction, regulation of morphology and trafficking, protein import and export, and other specialized processes. Discovery of these proteins by traditional biochemical fractionation can be plagued with contaminants and loss of key components. Using peroxidase-mediated proximity biotinylation, we captured and identified endogenous proteins on the outer mitochondrial membrane (OMM) and endoplasmic reticulum membrane (ERM) of living human fibroblasts. The proteomes of 137 and 634 proteins, respectively, are highly specific and highlight 94 potentially novel mitochondrial or ER proteins. Dataset intersection identified protein candidates potentially localized to mitochondria-ER contact sites. We found that one candidate, the tail-anchored, PDZ-domain-containing OMM protein SYNJ2BP, dramatically increases mitochondrial contacts with rough ER when overexpressed. Immunoprecipitation-mass spectrometry identified ribosome-binding protein 1 (RRBP1) as SYNJ2BP’s ERM binding partner. Our results highlight the power of proximity biotinylation to yield insights into the molecular composition and function of intracellular membranes. United States. National Institutes of Health (R01 CA186568) United States. National Institutes of Health (R01 GM077465) 2017-06-14T20:14:12Z 2017-06-14T20:14:12Z 2017-04 2016-12 Article http://purl.org/eprint/type/JournalArticle 2050-084X http://hdl.handle.net/1721.1/109872 Hung, Victoria; Lam, Stephanie S; Udeshi, Namrata D; Svinkina, Tanya; Guzman, Gaelen; Mootha, Vamsi K; Carr, Steven A andTing, Alice Y. “Proteomic Mapping of Cytosol-Facing Outer Mitochondrial and ER Membranes in Living Human Cells by Proximity Biotinylation.” eLife 6 (April 2017): e24463 © 2017 Hung et al https://orcid.org/0000-0003-3972-2820 https://orcid.org/0000-0002-2687-3470 https://orcid.org/0000-0002-7203-4299 https://orcid.org/0000-0002-8277-5226 en_US http://dx.doi.org/10.7554/eLife.24463 eLife Creative Commons Attribution 4.0 International License http://creativecommons.org/licenses/by/4.0/ application/pdf eLife Sciences Publications, Ltd. eLife |
| spellingShingle | Udeshi, Namrata D Svinkina, Tanya Hung, Victoria Lam, Stephanie Shih-Min Guzman, Gaelen Donnelly Mootha, Vamsi Carr, Steven A Ting, Alice Y Proteomic mapping of cytosol-facing outer mitochondrial and ER membranes in living human cells by proximity biotinylation |
| title | Proteomic mapping of cytosol-facing outer mitochondrial and ER membranes in living human cells by proximity biotinylation |
| title_full | Proteomic mapping of cytosol-facing outer mitochondrial and ER membranes in living human cells by proximity biotinylation |
| title_fullStr | Proteomic mapping of cytosol-facing outer mitochondrial and ER membranes in living human cells by proximity biotinylation |
| title_full_unstemmed | Proteomic mapping of cytosol-facing outer mitochondrial and ER membranes in living human cells by proximity biotinylation |
| title_short | Proteomic mapping of cytosol-facing outer mitochondrial and ER membranes in living human cells by proximity biotinylation |
| title_sort | proteomic mapping of cytosol facing outer mitochondrial and er membranes in living human cells by proximity biotinylation |
| url | http://hdl.handle.net/1721.1/109872 https://orcid.org/0000-0003-3972-2820 https://orcid.org/0000-0002-2687-3470 https://orcid.org/0000-0002-7203-4299 https://orcid.org/0000-0002-8277-5226 |
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