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복잡계자기조립연구단
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OrthoID: profiling dynamic proteomes through time and space using mutually orthogonal chemical tools

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Title
OrthoID: profiling dynamic proteomes through time and space using mutually orthogonal chemical tools
Author(s)
Ara Lee; Gihyun Sung; Sanghee Shin; Lee, Song-Yi; Jaehwan Sim; Nhung, Truong Thi My; Nghi, Tran Diem; Park, Sang Ki; Ponnusamy Pon Sathieshkumar; Kang, Imkyeung; Mun, Ji Young; Jong-Seo Kim; Rhee, Hyun-Woo; Park, Kyeng Min; Kimoon Kim
Publication Date
2024-02
Journal
Nature Communications, v.15, no.1
Publisher
Nature Publishing Group
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. © The Author(s) 2024.
URI
https://pr.ibs.re.kr/handle/8788114/15075
DOI
10.1038/s41467-024-46034-z
Appears in Collections:
Center for Self-assembly and Complexity(복잡계 자기조립 연구단) > 1. Journal Papers (저널논문)
Center for RNA Research(RNA 연구단) > 1. Journal Papers (저널논문)
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