Proteomic mapping of cytosol-facing outer mitochondrial and ER membranes in living human cells by proximity biotinylation
Name
Hung-2017-Proteomic mapping of cytosol-facing.pdf
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5.32 MB
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Author(s)
Udeshi, Namrata D
Svinkina, Tanya
Hung, Victoria
Lam, Stephanie Shih-Min
Guzman, Gaelen Donnelly
Mootha, Vamsi
Carr, Steven A
Ting, Alice Y
Date Issued
April 2017
Journal
eLife
Publisher
eLife Sciences Publications, Ltd.
Citation
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
Version
Final published version
Abstract
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.
MIT Department
Broad Institute of MIT and Harvard
Massachusetts Institute of Technology. Department of Chemistry
Terms of Use
Creative Commons Attribution 4.0 International License
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DOI of Published Version
https://doi.org/10.7554/eLife.24463
