Development of fluorescent dual-FRET probe for simultaneous detection of caspase-8 and caspase-9 activities and their relative quantification
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
36220004
DOI
10.1016/j.bioorg.2022.106151
PII: S0045-2068(22)00557-0
Knihovny.cz E-resources
- Keywords
- Caspases, FRET, Fluorescence, Fluorescent probe, Mathematical model, Proteases,
- MeSH
- Enzyme Activation MeSH
- Apoptosis MeSH
- Fluorescent Dyes * chemical synthesis chemistry MeSH
- Caspase 8 * analysis metabolism MeSH
- Caspase 9 * analysis metabolism MeSH
- Fluorescence Resonance Energy Transfer * methods MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Fluorescent Dyes * MeSH
- Caspase 8 * MeSH
- Caspase 9 * MeSH
A multi-FRET three-fluorophore probe containing coumarin, fluorescein and rhodamine B with two enzymatically cleavable linkers has been synthesized and optimized for the simultaneous activity detection and relative quantification of two proteases - caspase-8 and caspase-9. The probe designed as a ratiometric single-excitation triple-emission system shows specific change in fluorescence intensities upon enzymatic cleavage of individual linkers in model mixtures as well as in a cell lysate. The activation of caspase-8 and caspase-9 is responsible for initiation of extrinsic or intrinsic apoptotic pathway, respectively, and the probe was proposed as a single chemical tool which could help to decipher a mechanism of cell death induced by various stimuli. The main advantage of this probe is the simplicity of its preparation using conventional organic synthesis, easy application for measurement and evaluation of the results.
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