Mishra_2022_Chem.Sci_13_955

Reference

Title : A chemical tool for blue light-inducible proximity photo-crosslinking in live cells - Mishra_2022_Chem.Sci_13_955
Author(s) : Mishra PK , Kang MG , Lee H , Kim S , Choi S , Sharma N , Park CM , Ko J , Lee C , Seo JK , Rhee HW
Ref : Chem Sci , 13 :955 , 2022
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.

PubMedSearch : Mishra_2022_Chem.Sci_13_955
PubMedID:
Gene_locus related to this paper: rhoso-halo1

Related information

Inhibitor VL1-8MH    UL2-8MS
Gene_locus rhoso-halo1
Structure 7WAM    7WAN

Citations formats

Mishra PK, Kang MG, Lee H, Kim S, Choi S, Sharma N, Park CM, Ko J, Lee C, Seo JK, Rhee HW (2022)
A chemical tool for blue light-inducible proximity photo-crosslinking in live cells
Chem Sci 13 :955

Mishra PK, Kang MG, Lee H, Kim S, Choi S, Sharma N, Park CM, Ko J, Lee C, Seo JK, Rhee HW (2022)
Chem Sci 13 :955