6U2M

Crystal structure of a HaloTag-based calcium indicator, HaloCaMP V2, bound to JF635

6U2M の概要

• エントリーDOI: 10.2210/pdb6u2m/pdb
• 分子名称: HaloCaMP V2, (1E,3S)-1-{10-[2-carboxy-5-({2-[2-(hexyloxy)ethoxy]ethyl}carbamoyl)phenyl]-7-(3-fluoroazetidin-1-yl)-5,5-dimethyldibenz o[b,e]silin-3(5H)-ylidene}-3-fluoroazetidin-1-ium, CHLORIDE ION, ... (5 entities in total)
• 機能のキーワード: halotag, calcium, sensor, fluorescent, hydrolase
• 由来する生物種: Rhodococcus sp.
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 113499.40

構造登録者

Deo, C.,Schreiter, E.R. (登録日: 2019-08-20, 公開日: 2020-09-30, 最終更新日: 2023-10-11)

主引用文献

Deo, C.,Abdelfattah, A.S.,Bhargava, H.K.,Berro, A.J.,Falco, N.,Farrants, H.,Moeyaert, B.,Chupanova, M.,Lavis, L.D.,Schreiter, E.R.
The HaloTag as a general scaffold for far-red tunable chemigenetic indicators.
Nat.Chem.Biol., 17:718-723, 2021

PubMed Abstract: Functional imaging using fluorescent indicators has revolutionized biology, but additional sensor scaffolds are needed to access properties such as bright, far-red emission. Here, we introduce a new platform for 'chemigenetic' fluorescent indicators, utilizing the self-labeling HaloTag protein conjugated to environmentally sensitive synthetic fluorophores. We solve a crystal structure of HaloTag bound to a rhodamine dye ligand to guide engineering efforts to modulate the dye environment. We show that fusion of HaloTag with protein sensor domains that undergo conformational changes near the bound dye results in large and rapid changes in fluorescence output. This generalizable approach affords bright, far-red calcium and voltage sensors with highly tunable photophysical and chemical properties, which can reliably detect single action potentials in cultured neurons.

PubMed: 33795886
DOI: 10.1038/s41589-021-00775-w

実験手法

X-RAY DIFFRACTION (2 Å)