Cyclization Reaction-Based Turn-on Probe for Covalent Labeling of Target Proteins
Cyclization Reaction-Based Turn-on Probe for Covalent Labeling of Target Proteins
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DOI：
10.1016/j.chembiol.2020.01.006
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发表时间：
2020-03-19
影响因子：
8.6
通讯作者：
Itoh, Toshimasa
中科院分区：
文献类型：
--
作者：
Kojima, Hiroyuki;Fujita, Yuki;Itoh, Toshimasa
Fluorescent molecules have contributed to basic biological research but there are currently only a limited number of probes available for the detection of non-enzymatic proteins. Here, we report turn-on fluorescent probes mediated by conjugate addition and cyclization (TCC probes). These probes react with multiple amino acids and exhibit a 36-fold greater emission intensity after reaction. We analyzed the reactions between TCC probes and nuclear receptors by electrospray ionization mass spectrometry, X-ray crystallography, spectrofluorometry, and fluorescence microscopy. In vitro analysis showed that probes consisting of a protein ligand and TCC could label vitamin D receptor and peroxisome proliferator-activated receptor gamma. Moreover, we demonstrated that not only a ligand unit but also a peptide unit can label the target protein in a complex mixture.
