7WAM

Crystal structure of HaloTag complexed with VL1

Summary for 7WAM

• Entry DOI: 10.2210/pdb7wam/pdb
• Descriptor: Haloalkane dehalogenase, 3-[6-(2-azanylhydrazinyl)-1,3-bis(oxidanylidene)benzo[de]isoquinolin-2-yl]-N-[2-(2-hexoxyethoxy)ethyl]propanamide, CHLORIDE ION, ... (4 entities in total)
• Functional Keywords: halotag, haloalkane dehalogenase, hydrolase
• Biological source: Rhodococcus sp.
• Total number of polymer chains: 1
• Total formula weight: 34237.43

Authors

Pratyush, M.,Kang, M.,Lee, H.,Lee, C.,Rhee, H. (deposition date: 2021-12-14, release date: 2022-02-02, Last modification date: 2023-11-29)

Primary citation

Mishra, P.K.,Kang, M.G.,Lee, H.,Kim, S.,Choi, S.,Sharma, N.,Park, C.M.,Ko, J.,Lee, C.,Seo, J.K.,Rhee, H.W.
A chemical tool for blue light-inducible proximity photo-crosslinking in live cells.
Chem Sci, 13:955-966, 2022

PubMed Abstract: We developed a proximity photo-crosslinking method () with a 4-azido--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 , 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 -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.

PubMed: 35211260
DOI: 10.1039/d1sc04871f

Experimental method

X-RAY DIFFRACTION (1.49 Å)