Contact-ID, a tool for profiling organelle contact sites, reveals regulatory proteins of mitochondrial-associated membrane formation
- 15 May 2020
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 117 (22), 12109-12120
- https://doi.org/10.1073/pnas.1916584117
Abstract
The mitochondria-associated membrane (MAM) has emerged as a cellular signaling hub regulating various cellular processes. However, its molecular components remain unclear owing to lack of reliable methods to purify the intact MAM proteome in a physiological context. Here, we introduce Contact-ID, a split-pair system of BioID with strong activity, for identification of the MAM proteome in live cells. Contact-ID specifically labeled proteins proximal to the contact sites of the endoplasmic reticulum (ER) and mitochondria, and thereby identified 115 MAM-specific proteins. The identified MAM proteins were largely annotated with the outer mitochondrial membrane (OMM) and ER membrane proteins with MAM-related functions: e.g., FKBP8, an OMM protein, facilitated MAM formation and local calcium transport at the MAM. Furthermore, the definitive identification of biotinylation sites revealed membrane topologies of 85 integral membrane proteins. Contact-ID revealed regulatory proteins for MAM formation and could be reliably utilized to profile the proteome at any organelle–membrane contact sites in live cells.Keywords
Funding Information
- National Research Foundation of Korea (NRF-2019R1A2C3008463)
- National Research Foundation of Korea (NRF-2017R1A5A1015366)
- Ministry of Science, ICT and Future Planning (IBS-R008-D1)
- Ministry of Science, ICT and Future Planning (19-BR-01-08)
- National Research Foundation of Korea (2017R1A2B2009031)
- Samsung (SSTF-BA1601-14)
- Samsung (SSTF-BA1401-11)
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