Contact-ID, a tool for profiling organelle contact sites, reveals regulatory proteins of mitochondrial-associated membrane formation
- Authors
- Kwak, Chulhwan; Shin, Sanghee; Park, Jong-Seok; Jung, Minkyo; Nhung, Truong Thi My; Kang, Myeong-Gyun; Lee, Chaiheon; Kwon, Tae-Hyuk; Park, Sang Ki; Mun, Ji Young; Kim, Jong-Seo; Rhee, Hyun-Woo
- Issue Date
- Jun-2020
- Publisher
- NATL ACAD SCIENCES
- Keywords
- mitochondria-associated membrane (MAM); membrane contact site; proximity labeling; membrane protein topology; FKBP8
- Citation
- PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, v.117, no.22, pp.12109 - 12120
- Journal Title
- PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
- Volume
- 117
- Number
- 22
- Start Page
- 12109
- End Page
- 12120
- URI
- http://scholarworks.bwise.kr/kbri/handle/2023.sw.kbri/608
- DOI
- 10.1073/pnas.1916584117
- ISSN
- 0027-8424
- 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.
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