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

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Mishra, Pratyush Kumar; 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
Royal Society of Chemistry
Chemical Science, Vol.13 No.4, pp.955-966
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 bluelight-emitting diode light. Using an AzNP-conjugated HaloTag ligand (VL1), blue light-induced photocrosslinked 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.
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College of Natural Sciences (자연과학대학)Dept. of Chemistry (화학부)Journal Papers (저널논문_화학부)
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