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Development of Fluorogenic Probes for Rapid High‐Contrast Imaging of Transient Nuclear Localization of Sirtuin 3
Author(s) -
Gao Jingchi,
Hori Yuichiro,
Shimomura Takashi,
Bordy Mathieu,
Hasserodt Jens,
Kikuchi Kazuya
Publication year - 2020
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.201900568
Subject(s) - sirt3 , sirtuin , chemistry , ligand (biochemistry) , live cell imaging , coumarin , fluorescence , biophysics , fluorescence lifetime imaging microscopy , nuclear localization sequence , intracellular , biochemistry , cell , biology , acetylation , receptor , cytoplasm , physics , organic chemistry , quantum mechanics , gene
Protein labeling using fluorogenic probes enables the facile visualization of proteins of interest. Herein, we report new fluorogenic probes consisting of a rationally designed coumarin ligand for the live‐cell fluorogenic labeling of the photoactive yellow protein (PYP)‐tag. On the basis of the photochemical mechanisms of coumarin and the probe–tag interactions, we introduced a hydroxy group into an environment‐sensitive coumarin ligand to modulate its spectroscopic properties and increase the labeling reaction rate. The resulting probe had a higher labeling reaction rate constant and a greater fluorescence OFF–ON ratio than any previously developed PYP‐tag labeling probe. The probe enabled the fluorogenic labeling of intracellular proteins within minutes. Furthermore, we used our probe to investigate the localization of sirtuin 3 (SIRT3), a mitochondrial deacetylase. Although the nuclear localization of SIRT3 has been controversial, this transient nuclear localization was clearly captured by the rapid, high‐contrast imaging enabled by our probe.