한빛사논문
- 조회280
Ara Lee 1,2,12, Gihyun Sung 1,2,12, Sanghee Shin 3,4, Song-Yi Lee 5, Jaehwan Sim 1,6, Truong Thi My Nhung 7, Tran Diem Nghi 7, Sang Ki Park 7, Ponnusamy Pon Sathieshkumar 1, Imkyeung Kang 8,9, Ji Young Mun 8, Jong-Seo Kim 3,4,*, Hyun-Woo Rhee 5,*, Kyeng Min Park 10,* & Kimoon Kim 1,2,6,11,*
1Center for Self–assembly and Complexity, Institute for Basic Science (IBS), Pohang, Republic of Korea.
2Division of Advanced Materials Science (AMS), Pohang University of Science and Technology (POSTECH), Pohang, Republic of Korea.
3Center for RNA Research, Institute for Basic Science (IBS), Seoul, Republic of Korea.
4School of Biological Sciences, Seoul National University, Seoul, Republic of Korea.
5Department of Chemistry, Seoul National University, Seoul, Republic of Korea.
6School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology (POSTECH), Pohang, Republic of Korea.
7Department of Life Sciences, Pohang University of Science and Technology (POSTECH), Pohang, Republic of Korea.
8Neural Circuit Research Group, Korea Brain Research Institute, Daegu, Republic of Korea.
9Department of Microbiology, University of Ulsan College of Medicine, Ulsan, Republic of Korea.
10Department of Biochemistry, Daegu Catholic University School of Medicine, Daegu, Republic of Korea.
11Department of Chemistry, Pohang University of Science and Technology (POSTECH), Pohang, Republic of Korea.
12These authors contributed equally: Ara Lee, Gihyun Sung.
*Corresponding authors: correspondence to Jong-Seo Kim, Hyun-Woo Rhee, Kyeng Min Park or Kimoon Kim
Abstract
Identifying proteins at organelle contact sites, such as mitochondria-associated endoplasmic reticulum membranes (MAM), is essential for understanding vital cellular processes, yet challenging due to their dynamic nature. Here we report “OrthoID”, a proteomic method utilizing engineered enzymes, TurboID and APEX2, for the biotinylation (Bt) and adamantylation (Ad) of proteins close to the mitochondria and endoplasmic reticulum (ER), respectively, in conjunction with high-affinity binding pairs, streptavidin-biotin (SA-Bt) and cucurbit[7]uril-adamantane (CB[7]-Ad), for selective orthogonal enrichment of Bt- and Ad-labeled proteins. This approach effectively identifies protein candidates associated with the ER-mitochondria contact, including LRC59, whose roles at the contact site were—to the best of our knowledge—previously unknown, and tracks multiple protein sets undergoing structural and locational changes at MAM during mitophagy. These findings demonstrate that OrthoID could be a powerful proteomics tool for the identification and analysis of spatiotemporal proteins at organelle contact sites and revealing their dynamic behaviors in vital cellular processes.
논문정보
- Research article
- 2024년 02월 (BRIC 등록일 2024-03-04)
- 국내 연구진
- 바이오·의료융합 > 바이오센싱 및 나노바이오물질
관련 보도자료
- Bio통신원
- 2024. 04. 03
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