7ZIZ

X-ray structure of the dead variant haloalkane dehalogenase HaloTag7-D106A bound to a pentanol tetramethylrhodamine ligand (TMR-Hy5)

7ZIZ の概要

• エントリーDOI: 10.2210/pdb7ziz/pdb
• 関連するPDBエントリー: 7ZIV 7ZIW 7ZIX 7ZIY 7ZJ0
• 分子名称: Haloalkane dehalogenase, [9-[2-carboxy-5-[2-[2-(5-oxidanylpentoxy)ethoxy]ethylcarbamoyl]phenyl]-6-(dimethylamino)xanthen-3-ylidene]-dimethyl-azanium, CHLORIDE ION, ... (5 entities in total)
• 機能のキーワード: haloalkane dehalogenase, halotag, halotag7, self-labeling protein, fluorophore, tetramethylrhodamine, hydrolase
• 由来する生物種: Rhodococcus sp.
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 33914.23

構造登録者

Tarnawski, M.,Kompa, J.,Johnsson, K.,Hiblot, J. (登録日: 2022-04-08, 公開日: 2023-02-22, 最終更新日: 2024-02-07)

主引用文献

Kompa, J.,Bruins, J.,Glogger, M.,Wilhelm, J.,Frei, M.S.,Tarnawski, M.,D'Este, E.,Heilemann, M.,Hiblot, J.,Johnsson, K.
Exchangeable HaloTag Ligands for Super-Resolution Fluorescence Microscopy.
J.Am.Chem.Soc., 145:3075-3083, 2023

PubMed Abstract: The specific and covalent labeling of the protein HaloTag with fluorescent probes in living cells makes it a powerful tool for bioimaging. However, the irreversible attachment of the probe to HaloTag precludes imaging applications that require transient binding of the probe and comes with the risk of irreversible photobleaching. Here, we introduce exchangeable ligands for fluorescence labeling of HaloTag (xHTLs) that reversibly bind to HaloTag and that can be coupled to rhodamines of different colors. In stimulated emission depletion (STED) microscopy, probe exchange of xHTLs allows imaging with reduced photobleaching as compared to covalent HaloTag labeling. Transient binding of fluorogenic xHTLs to HaloTag fusion proteins enables points accumulation for imaging in nanoscale topography (PAINT) and MINFLUX microscopy. We furthermore introduce pairs of xHTLs and HaloTag mutants for dual-color PAINT and STED microscopy. xHTLs thus open up new possibilities in imaging across microscopy platforms for a widely used labeling approach.

PubMed: 36716211
DOI: 10.1021/jacs.2c11969

実験手法

X-RAY DIFFRACTION (1.5 Å)