Premium
Ratiometric Detection of Glutathione Based on Disulfide Linkage Rupture between a FRET Coumarin Donor and a Rhodamine Acceptor
Author(s) -
Zhang Yibin,
Xia Shuai,
Wan Shulin,
Steenwinkel Tessa E.,
Vohs Tara,
Luck Rudy L.,
Werner Thomas,
Liu Haiying
Publication year - 2021
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.202100108
Subject(s) - förster resonance energy transfer , glutathione , fluorescence , rhodamine , chemistry , photochemistry , acceptor , cysteine , coumarin , hela , biophysics , biochemistry , organic chemistry , in vitro , physics , quantum mechanics , biology , enzyme , condensed matter physics
Abstract Abnormal levels of glutathione, a cellular antioxidant, can lead to a variety of diseases. We have constructed a near‐infrared ratiometric fluorescent probe to detect glutathione concentrations in biological samples. The probe consists of a coumarin donor, which is connected through a disulfide‐tethered linker to a rhodamine acceptor. Under the excitation of the coumarin donor at 405 nm, the probe shows weak visible fluorescence of the coumarin donor at 470 nm and strong near‐infrared fluorescence of the rhodamine acceptor at 652 nm due to efficient Forster resonance energy transfer (FRET) from the donor to the acceptor. Glutathione breaks the disulfide bond through reduction, which results in a dramatic increase in coumarin fluorescence and a corresponding decrease in rhodamine fluorescence. The probe possesses excellent cell permeability, biocompatibility, and good ratiometric fluorescence responses to glutathione and cysteine with a self‐calibration capability. The probe was utilized to ratiometrically visualize glutathione concentration alterations in HeLa cells and Drosophila melanogaster larvae.