Improvement of the Green-Red Förster Resonance Energy Transfer-Based Ca²⁺ Indicator by Using the Green Fluorescent Protein, Gamillus, with a Trans Chromophore as the Donor

Matsuda, Tomoki; Sakai, Shinya; Okazaki, Kei-Ichi et al.

ACS Sensors, 2024, 9(4), 1743-1748

Number of Access:635（2025-11-02 23:05 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/95641

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Title
Improvement of the Green-Red Förster Resonance Energy Transfer-Based Ca²⁺ Indicator by Using the Green Fluorescent Protein, Gamillus, with a Trans Chromophore as the Donor
Creator
NRID
1000050419206 1000050419206
researchmap permalink
tikimaz tikimaz
Scopus Author ID
7404349300 7404349300
CreatorName
Matsuda, Tomoki
CreatorName
Sakai, Shinya
NRID
1000050792529 1000050792529
CreatorName
Okazaki, Kei-Ichi
NRID
1000020311350 1000020311350
ORCID
0000-0003-2650-9895 0000-0003-2650-9895
researchmap permalink
ng1 ng1
Scopus Author ID
7403433377 7403433377
CreatorName
Nagai, Takeharu
Subject
fluorescent indicator
fluorescent protein
Förster resonance energy transfer
genetically encoded Ca2+ indicators
protein engineering
Description
Matsuda T., Sakai S., Okazaki K.I., et al. Improvement of the Green-Red Förster Resonance Energy Transfer-Based Ca²⁺ Indicator by Using the Green Fluorescent Protein, Gamillus, with a Trans Chromophore as the Donor. ACS Sensors 9, 1743 (2024); https://doi.org/10.1021/acssensors.3c02398.
Abstract
To monitor the Ca²⁺ dynamics in cells, various genetically encoded Ca²⁺ indicators (GECIs) based on Förster resonance energy transfer (FRET) between fluorescent proteins are widely used for live imaging. Conventionally, cyan and yellow fluorescent proteins have been often used as FRET pairs. Meanwhile, bathochromically shifted indicators with green and red fluorescent protein pairs have various advantages, such as low toxicity and autofluorescence in cells. However, it remains difficult to develop them with a similar level of dynamic range as cyan and yellow fluorescent protein pairs. To improve this, we used Gamillus, which has a unique trans-configuration chromophore, as a green fluorescent protein. Based on one of the best high-dynamic-range GECIs, Twitch-NR, we developed a GECI with 1.5-times higher dynamic range (253%), Twitch-GmRR, using RRvT as a red fluorescent protein. Twitch-GmRR had high brightness and photostability and was successfully applied for imaging the Ca²⁺ dynamics in live cells. Our results suggest that Gamillus with trans-type chromophores contributes to improving the dynamic range of GECIs. Therefore, selection of the cis-trans isomer of the chromophore may be a fundamental approach to improve the dynamic range of green-red FRET indicators, unlimited by GECIs.
Publisher
American Chemical Society
Source Title
ACS Sensors
Volume (Issue)
9(4)
Page
1743-1748
Date of Issued
2024-04-26
Language
English
Handle URL
https://hdl.handle.net/11094/95641
eISSN
23793694
Relation.isIdenticalTo
DOI (Publisher Version)
https://doi.org/10.1021/acssensors.3c02398
Access Rights
open access
CopyRight
This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
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Version of Record
Category
学術雑誌論文 Journal Article