한빛사논문
- 조회3,395
Chulhwan Kwaka,b,1, Sanghee Shinc,d,1, Jong-Seok Parkb,1, Minkyo Junge, Truong Thi My Nhungf, Myeong-Gyun Kanga,b, Chaiheon Leeb, Tae-Hyuk Kwonb, Sang Ki Parkf,2, Ji Young Mune,2, Jong-Seo Kimc,d,2, and Hyun-Woo Rheea,d,2
aDepartment of Chemistry, Seoul National University, 08826 Seoul, Korea; bDepartment of Chemistry, Ulsan National Institute of Science and Technology, 44919 Ulsan, Korea; cCenter for RNA Research, Institute for Basic Science, Seoul 08826, Korea; dSchool of Biological Sciences, Seoul National University, 08826 Seoul, Korea; eNeural Circuit Research Group, Korea Brain Research Institute, 41062 Daegu, Korea; and fDepartment of Life Sciences, Pohang University of Science and Technology, 37673 Pohang, Korea
1C.K., S.S., and J.-S.P. contributed equally to this work.
2To whom correspondence may be addressed.
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.
mitochondria-associated membrane (MAM), membrane contact site, proximity labeling, membrane protein topology, FKBP8
논문정보
TOP52020년 후보
- Research article
- 2020년 05월 (BRIC 등록일 2020-05-20)
- 국내 연구진
- 생명과학 > 생화학
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