Department of Organic Chemistry, Faculty of Science, Palacký University, 17. listopadu 12, 771 46 Olomouc, Czech Republic.
Department of Experimental Biology, Faculty of Science, Palacký University, Šlechtitelů 27, 783 71 Olomouc, Czech Republic.
Bioorg Chem. 2022 Dec;129:106151. doi: 10.1016/j.bioorg.2022.106151. Epub 2022 Sep 28.
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.
一种含有香豆素、荧光素和若丹明 B 的多 FRET 三荧光探针，带有两个酶切连接子，已被合成并优化，用于同时检测和相对定量两种蛋白酶 - 半胱天冬酶-8 和半胱天冬酶-9。该探针设计为比率型单激发三发射系统，在模型混合物以及细胞裂解物中，单个连接子的酶切会导致荧光强度发生特异性变化。半胱天冬酶-8 和半胱天冬酶-9 的激活分别负责外源性或内源性凋亡途径的启动，该探针被提出作为一种单一的化学工具，可以帮助破译各种刺激诱导的细胞死亡的机制。该探针的主要优点是其制备方法简单，使用常规有机合成，易于测量和评估结果。
