Switching Between Ultrafast Pathways Enables a Green-Red Emission Ratiometric Fluorescent-Protein-Based Ca Biosensor

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2021 Jan 20

PMID 33466257

Citations 6

Authors

Longteng Tang

Shuce Zhang

Yufeng Zhao

Nikita D Rozanov

Liangdong Zhu

Jiahui Wu

Robert E Campbell

Chong Fang

Affiliations

Soon will be listed here.

Abstract

Ratiometric indicators with long emission wavelengths are highly preferred in modern bioimaging and life sciences. Herein, we elucidated the working mechanism of a standalone red fluorescent protein (FP)-based Ca biosensor, REX-GECO1, using a series of spectroscopic and computational methods. Upon 480 nm photoexcitation, the Ca-free biosensor chromophore becomes trapped in an excited dark state. Binding with Ca switches the route to ultrafast excited-state proton transfer through a short hydrogen bond to an adjacent Glu80 residue, which is key for the biosensor's functionality. Inspired by the 2D-fluorescence map, REX-GECO1 for Ca imaging in the ionomycin-treated human HeLa cells was achieved for the first time with a red/green emission ratio change (ΔR/R) of ~300%, outperforming many FRET- and single FP-based indicators. These spectroscopy-driven discoveries enable targeted design for the next-generation biosensors with larger dynamic range and longer emission wavelengths.

Citing Articles

Delineating Ultrafast Structural Dynamics of a Green-Red Fluorescent Protein for Calcium Sensing.

Krueger T, Tang L, Fang C Biosensors (Basel). 2023; 13(2).

PMID: 36831983 PMC: 9954042. DOI: 10.3390/bios13020218.

Rational Design for High Bioorthogonal Fluorogenicity of Tetrazine-Encoded Green Fluorescent Proteins.

Tang L, Bednar R, Rozanov N, Hemshorn M, Mehl R, Fang C Nat Sci (Weinh). 2022; 2(4).

PMID: 36440454 PMC: 9699285. DOI: 10.1002/ntls.20220028.

Encoding, transmission, decoding, and specificity of calcium signals in plants.

Allan C, Morris R, Meisrimler C J Exp Bot. 2022; 73(11):3372-3385.

PMID: 35298633 PMC: 9162177. DOI: 10.1093/jxb/erac105.

Application of Fluorescent Proteins for Functional Dissection of the Visual System.

Smylla T, Wagner K, Huber A Int J Mol Sci. 2021; 22(16).

PMID: 34445636 PMC: 8396179. DOI: 10.3390/ijms22168930.

Nanomaterial-Based Dual-Emission Ratiometric Fluorescent Sensors for Biosensing and Cell Imaging.

Zhang Y, Hou D, Wang Z, Cai N, Au C Polymers (Basel). 2021; 13(15).

PMID: 34372142 PMC: 8348892. DOI: 10.3390/polym13152540.

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