2M97

Optimized Ratiometric Calcium Sensors For Functional In Vivo Imaging of Neurons and T-Lymphocytes

2M97 の概要

• エントリーDOI: 10.2210/pdb2m97/pdb
• NMR情報: BMRB: 19285
• 分子名称: Optimized Ratiometric Calcium Sensor (1 entity in total)
• 機能のキーワード: calcium biosensors, metal binding protein
• 由来する生物種: Opsanus tau
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 8066.65

構造登録者

Russo, L.,Becker, S.,Griesinger, C. (登録日: 2013-06-04, 公開日: 2014-01-01, 最終更新日: 2024-05-01)

主引用文献

Thestrup, T.,Litzlbauer, J.,Bartholomaus, I.,Mues, M.,Russo, L.,Dana, H.,Kovalchuk, Y.,Liang, Y.,Kalamakis, G.,Laukat, Y.,Becker, S.,Witte, G.,Geiger, A.,Allen, T.,Rome, L.C.,Chen, T.W.,Kim, D.S.,Garaschuk, O.,Griesinger, C.,Griesbeck, O.
Optimized ratiometric calcium sensors for functional in vivo imaging of neurons and T lymphocytes.
Nat.Methods, 11:175-182, 2014

PubMed Abstract: The quality of genetically encoded calcium indicators (GECIs) has improved dramatically in recent years, but high-performing ratiometric indicators are still rare. Here we describe a series of fluorescence resonance energy transfer (FRET)-based calcium biosensors with a reduced number of calcium binding sites per sensor. These 'Twitch' sensors are based on the C-terminal domain of Opsanus troponin C. Their FRET responses were optimized by a large-scale functional screen in bacterial colonies, refined by a secondary screen in rat hippocampal neuron cultures. We tested the in vivo performance of the most sensitive variants in the brain and lymph nodes of mice. The sensitivity of the Twitch sensors matched that of synthetic calcium dyes and allowed visualization of tonic action potential firing in neurons and high resolution functional tracking of T lymphocytes. Given their ratiometric readout, their brightness, large dynamic range and linear response properties, Twitch sensors represent versatile tools for neuroscience and immunology.

PubMed: 24390440
DOI: 10.1038/nmeth.2773

実験手法

SOLUTION NMR