8B6T

X-ray structure of the interface optimized haloalkane dehalogenase HaloTag7 fusion to the green fluorescent protein GFP (ChemoG5-TMR) labeled with a chloroalkane tetramethylrhodamine fluorophore substrate

8B6T の概要

• エントリーDOI: 10.2210/pdb8b6t/pdb
• 関連するPDBエントリー: 8B6R 8B6S
• 分子名称: Green fluorescent protein,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)
• 機能のキーワード: haloalkane dehalogenase, halotag, halotag7, self-labeling protein, green fluorescent protein, gfp, fluorophore, tetramethylerhodamine, tmr, hydrolase
• 由来する生物種: Rhodococcus sp.; 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 121699.88

構造登録者

Tarnawski, M.,Hellweg, L.,Hiblot, J. (登録日: 2022-09-27, 公開日: 2023-07-26, 最終更新日: 2024-10-09)

主引用文献

Hellweg, L.,Edenhofer, A.,Barck, L.,Huppertz, M.C.,Frei, M.S.,Tarnawski, M.,Bergner, A.,Koch, B.,Johnsson, K.,Hiblot, J.
A general method for the development of multicolor biosensors with large dynamic ranges.
Nat.Chem.Biol., 19:1147-1157, 2023

PubMed Abstract: Fluorescent biosensors enable the study of cell physiology with spatiotemporal resolution; yet, most biosensors suffer from relatively low dynamic ranges. Here, we introduce a family of designed Förster resonance energy transfer (FRET) pairs with near-quantitative FRET efficiencies based on the reversible interaction of fluorescent proteins with a fluorescently labeled HaloTag. These FRET pairs enabled the straightforward design of biosensors for calcium, ATP and NAD with unprecedented dynamic ranges. The color of each of these biosensors can be readily tuned by changing either the fluorescent protein or the synthetic fluorophore, which enables simultaneous monitoring of free NAD in different subcellular compartments following genotoxic stress. Minimal modifications of these biosensors furthermore allow their readout to be switched to fluorescence intensity, fluorescence lifetime or bioluminescence. These FRET pairs thus establish a new concept for the development of highly sensitive and tunable biosensors.

PubMed: 37291200
DOI: 10.1038/s41589-023-01350-1

実験手法

X-RAY DIFFRACTION (2 Å)