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A chemical tool for blue light-inducible proximity photo-crosslinking in live cells

Title
A chemical tool for blue light-inducible proximity photo-crosslinking in live cells
Authors
Mishra, Pratyush MishraKang, Myeong-GyunLee, HakbongKim, SeungjoonChoi, SubinSharma, NirmaliPark, Cheol-MinKo, JaewonLee, ChangwookSeo, Jeong KonRhee, Hyun-Woo
DGIST Authors
Mishra, Pratyush Mishra; Kang, Myeong-Gyun; Lee, Hakbong; Kim, Seungjoon; Choi, Subin; Sharma, Nirmali; Park, Cheol-Min; Ko, Jaewon; Lee, Changwook; Seo, Jeong Kon; Rhee, Hyun-Woo
Issue Date
2022-01
Citation
Chemical Science, 13(4), 955-966
Type
Article
Keywords
HALOTAG7REVEALSSTAUFENLIGASEPROBESPROTEIN-PROTEIN INTERACTIONSHYDROGEN-SULFIDEARYL AZIDETAGBINDING
ISSN
2041-6520
Abstract
We developed a proximity photo-crosslinking method (Spotlight) with a 4-azido-N-ethyl-1,8-naphthalimide (AzNP) moiety that can be converted to reactive aryl nitrene species using ambient blue light-emitting diode light. Using an AzNP-conjugated HaloTag ligand (VL1), blue light-induced photo-crosslinked products of various HaloTag-conjugated proteins of interest were detected in subcellular spaces in live cells. Chemical or heat stress-induced dynamic changes in the proteome were also detected, and photo-crosslinking in the mouse brain tissue was enabled. Using Spotlight, we further identified the host interactome of SARS-CoV-2 nucleocapsid (N) protein, which is essential for viral genome assembly. Mass analysis of the VL1-crosslinked product of N-HaloTag in HEK293T cells showed that RNA-binding proteins in stress granules were exclusively enriched in the cross-linked samples. These results tell that our method can reveal the interactome of protein of interest within a short distance in live cells. © 2022 The Royal Society of Chemistry.
URI
http://hdl.handle.net/20.500.11750/16041
DOI
10.1039/d1sc04871f
Publisher
Royal Society of Chemistry
Related Researcher
  • Author Ko, Jaewon Laboratory of Synapse Formation and Function
  • Research Interests Synapse Formation and Function; Neural Circuits; 뇌질환; animal model
Files:
Collection:
Department of Brain SciencesLaboratory of Synapse Formation and Function1. Journal Articles


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