7PCW

X-RAY STRUCTURE OF THE HALOALKANE DEHALOGENASE HALOTAG7-M175W LABELED WITH A CHLOROALKANE-TETRAMETHYLRHODAMINE FLUOROPHORE SUBSTRATE

Summary for 7PCW

• Entry DOI: 10.2210/pdb7pcw/pdb
• Related: 6ZVU 6ZVV 6ZVW 6ZVX 6ZVY 7PCX
• Descriptor: Haloalkane dehalogenase, [9-[2-carboxy-5-[2-[2-(6-chloranylhexoxy)ethoxy]ethylcarbamoyl]phenyl]-6-(dimethylamino)xanthen-3-ylidene]-dimethyl-azanium, CHLORIDE ION, ... (4 entities in total)
• Functional Keywords: haloalkane dehalogenase, halo, tag, halotag7, self-labeling protein, fluorophore, tetramethylrhodamine, hydrolase
• Biological source: Rhodococcus sp.
• Total number of polymer chains: 4
• Total formula weight: 135814.54

Authors

Tarnawski, M.,Frei, M.,Hiblot, J.,Johnsson, K. (deposition date: 2021-08-04, release date: 2021-12-15, Last modification date: 2024-11-13)

Primary citation

Frei, M.S.,Tarnawski, M.,Roberti, M.J.,Koch, B.,Hiblot, J.,Johnsson, K.
Engineered HaloTag variants for fluorescence lifetime multiplexing.
Nat.Methods, 19:65-70, 2022

PubMed Abstract: Self-labeling protein tags such as HaloTag are powerful tools that can label fusion proteins with synthetic fluorophores for use in fluorescence microscopy. Here we introduce HaloTag variants with either increased or decreased brightness and fluorescence lifetime compared with HaloTag7 when labeled with rhodamines. Combining these HaloTag variants enabled live-cell fluorescence lifetime multiplexing of three cellular targets in one spectral channel using a single fluorophore and the generation of a fluorescence lifetime-based biosensor. Additionally, the brightest HaloTag variant showed up to 40% higher brightness in live-cell imaging applications.

PubMed: 34916672
DOI: 10.1038/s41592-021-01341-x

Experimental method

X-RAY DIFFRACTION (2.3 Å)