Development of fluorescent dual-FRET probe for simultaneous detection of caspase-8 and caspase-9 activities and their relative quantification
Jazyk angličtina Země Spojené státy americké Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
36220004
DOI
10.1016/j.bioorg.2022.106151
PII: S0045-2068(22)00557-0
Knihovny.cz E-zdroje
- Klíčová slova
- Caspases, FRET, Fluorescence, Fluorescent probe, Mathematical model, Proteases,
- MeSH
- aktivace enzymů MeSH
- apoptóza MeSH
- fluorescenční barviva * chemická syntéza chemie MeSH
- kaspasa 8 * analýza metabolismus MeSH
- kaspasa 9 * analýza metabolismus MeSH
- rezonanční přenos fluorescenční energie * metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- fluorescenční barviva * MeSH
- kaspasa 8 * MeSH
- kaspasa 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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