Displacement Induced Off-On Fluorescent Biosensor Targeting IDO1 Activity in Live Cells.
Anal Chem
; 91(23): 14943-14950, 2019 12 03.
Article
em En
| MEDLINE
| ID: mdl-31714063
ABSTRACT
We show how the macrocyclic host cucurbit[8]uril (CB[8]) and a fluorescent dye form a biosensing ensemble while its cavity simultaneously traps tryptophan, the upstream substrate of IDO1 enzymes, therefore providing a label-free method to monitor the activity of IDO1 in real time. Incubation of malignant HeLa and HepG2 cells overexpressing IDO1 with the associative biosensor resulted in its spontaneous uptake and a fluorescence switch-on response in situ, which can be traced to the displacement of tryptophan from CB[8] upon IDO1-catalyzed oxidation. The results, for the first time, establish a supramolecular sensing concept for the detection of intracellular enzymatic activity in live cells, thus allowing direct cell-based analysis and inhibitor screening compatible with commercial instruments including microplate reader, fluorescent microscopy, and flow cytometry.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Técnicas Biossensoriais
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Indolamina-Pirrol 2,3,-Dioxigenase
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Corantes Fluorescentes
Limite:
Humans
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article