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Publication Abstract from PubMed

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.

A chemical tool for blue light-inducible proximity photo-crosslinking in live cells.,Mishra PK, Kang MG, Lee H, Kim S, Choi S, Sharma N, Park CM, Ko J, Lee C, Seo JK, Rhee HW Chem Sci. 2021 Dec 10;13(4):955-966. doi: 10.1039/d1sc04871f. eCollection 2022, Jan 26. PMID:35211260^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.