7ZJ0

X-ray structure of the haloalkane dehalogenase HaloTag7 bound to a pentylmethanesulfonamide tetramethylrhodamine ligand (TMR-S5)

Summary for 7ZJ0

• Entry DOI: 10.2210/pdb7zj0/pdb
• Related: 7ZIV 7ZIW 7ZIX 7ZIY 7ZIZ
• Descriptor: Haloalkane dehalogenase, [9-[2-carboxy-5-[2-[2-[5-(methylsulfonylamino)pentoxy]ethoxy]ethylcarbamoyl]phenyl]-6-(dimethylamino)xanthen-3-ylidene]-dimethyl-azanium, GLYCEROL, ... (4 entities in total)
• Functional Keywords: haloalkane dehalogenase, halotag, halotag7, self-labeling protein, fluorophore, tetramethylrhodamine, hydrolase
• Biological source: Rhodococcus sp.
• Total number of polymer chains: 2
• Total formula weight: 68368.16

Authors

Tarnawski, M.,Kompa, J.,Johnsson, K.,Hiblot, J. (deposition date: 2022-04-08, release date: 2023-02-22, Last modification date: 2024-02-07)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.5 Å)