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Developing Bright Green Fluorescent Protein (GFP)‐like Fluorogens for Live‐Cell Imaging with Nonpolar Protein−Chromophore Interactions
Author(s) -
Chen Cheng,
Tachibana Sean R.,
Baleeva Nadezhda S.,
Myasnyanko Ivan N.,
Bogdanov Alexey M.,
Gavrikov Alexey S.,
Mishin Alexander S.,
Malyshevskaya Kseniya K.,
Baranov Mikhail S.,
Fang Chong
Publication year - 2021
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.202101250
Subject(s) - chromophore , fluorescence , green fluorescent protein , rational design , chemistry , biophysics , fluorescent protein , live cell imaging , phototoxicity , fluorescence lifetime imaging microscopy , photochemistry , nanotechnology , materials science , cell , optics , biochemistry , biology , physics , in vitro , gene
Fluorescence‐activating proteins (FAPs) that bind a chromophore and activate its fluorescence have gained popularity in bioimaging. The fluorescence‐activating and absorption‐shifting tag (FAST) is a light‐weight FAP that enables fast reversible fluorogen binding, thus advancing multiplex and super‐resolution imaging. However, the rational design of FAST‐specific fluorogens with large fluorescence enhancement (FE) remains challenging. Herein, a new fluorogen directly engineered from green fluorescent protein (GFP) chromophore by a unique double‐donor‐one‐acceptor strategy, which exhibits an over 550‐fold FE upon FAST binding and a high extinction coefficient of approximately 100,000 M −1 cm −1 , is reported. Correlation analysis of the excited state nonradiative decay rates and environmental factors reveal that the large FE is caused by nonpolar protein−fluorogen interactions. Our deep insights into structure‐function relationships could guide the rational design of bright fluorogens for live‐cell imaging with extended spectral properties such as redder emissions.