Dual-Mechanism Quenched Fluorogenic Probe Provides Selective and Rapid Detection of Cathepsin L Activity*.
Dual-Mechanism Quenched Fluorogenic Probe Provides Selective and Rapid Detection of Cathepsin L Activity*.
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DOI：
10.1002/cmdc.202000823
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发表时间：
2021-04-08
期刊：
影响因子：
3.4
通讯作者：
Cui L
中科院分区：
文献类型：
--
作者：
Schleyer KA;Fetrow B;Zannes Fatland P;Liu J;Chaaban M;Ma B;Cui L
组织蛋白酶L （CTL）是一种半胱氨酸蛋白酶，在许多疾病状态下表现出上调的活性。重叠底物特异性使得CTL活性的选择性检测难以从其相近的同源物CTV和普遍存在的CTB中进行分析。目前的CTL活性探针由于对比度差或需要额外的分析步骤来实现选择性而应用有限。我们开发了一种荧光探针CTLAP，它对CTB和CTV的CTL具有良好的选择性，同时由于双重猝灭机制而表现出低背景荧光。CTLAP在孵育的前10分钟内达到最佳CTL选择性，这表明它可以快速检测CTL，即使存在竞争性组织蛋白酶。一种新型的组织蛋白酶L激活探针（CTLAP）具有一种化学结构，可以同时为其目标提供高选择性，同时淬灭其自身的背景发射，从而在10分钟的培养时间内产生卓越的对比度，并在消耗时产生120倍的荧光。作为一种新型的检测工具，CTLAP揭示了化学结构可以影响这些工具的光物理性质以及选择性。
Cathepsin L (CTL) is a cysteine protease demonstrating upregulated activity in many disease states. Overlapping substrate specificity makes selective detection of CTL activity difficult to parse from its close homologue CTV and the ubiquitous CTB. Current probes of CTL activity have limited applications due to either poor contrast or extra assay steps required to achieve selectivity. We have developed a fluorogenic probe, CTLAP, which displays good selectivity for CTL over CTB and CTV while exhibiting low background fluorescence attributed to dual quenching mechanisms. CTLAP achieves optimum CTL selectivity in the first 10 min of incubation suggesting that it is amenable for rapid detection of CTL, even in the presence of competing cathepsins. A novel cathepsin L activable probe (CTLAP) bears a chemical structure that simultaneously provides high selectivity for its target while quenching its own background emission, resulting in superior contrast within 10 min of incubation time and 120-fold turn-on fluorescence upon consumption. Along with being a novel detection tool, CTLAP reveals that chemical structure can influence photophysical properties as well as the selectivity of such tools.
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