Coumarin-Based Profluorescent and Fluorescent Substrates for Determining Xenobiotic-Metabolizing Enzyme Activities In Vitro
in vivo methods, such as spectrophotometric, fluorometric, mass spectrometric,<br />and radioactivity-based techniques. In fluorescence-based assays, the reaction produces a fluorescent<br />product from a nonfluorescent substrate or vice versa. Fluorescence-based enzyme assays are<br...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-07-01
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| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/21/13/4708 |
| Summary: | in vivo methods, such as spectrophotometric, fluorometric, mass spectrometric,<br />and radioactivity-based techniques. In fluorescence-based assays, the reaction produces a fluorescent<br />product from a nonfluorescent substrate or vice versa. Fluorescence-based enzyme assays are<br />usually highly sensitive and specific, allowing measurements on small specimens of tissues with<br />low enzyme activities. Fluorescence assays are also amenable to miniaturization of the reaction<br />mixtures and can thus be done in high throughput. 7-Hydroxycoumarin and its derivatives are<br />widely used as fluorophores due to their desirable photophysical properties. They possess a large -<br />conjugated system with electron-rich and charge transfer properties. This conjugated structure leads<br />to applications of 7-hydroxycoumarins as fluorescent sensors for biological activities. We describe in<br />this review historical highlights and current use of coumarins and their derivatives in evaluating<br />activities of the major types of xenobiotic-metabolizing enzyme systems. Traditionally, coumarin<br />substrates have been used to measure oxidative activities of cytochrome P450 (CYP) enzymes. For this<br />purpose, profluorescent coumarins are very sensitive, but generally lack selectivity for individual CYP<br />forms. With the aid of molecular modeling, we have recently described several new coumarin-based<br />substrates for measuring activities of CYP and conjugating enzymes with improved selectivity. |
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| ISSN: | 1661-6596 1422-0067 |