Super-silent FRET Sensor Enables Live Cell Imaging and Flow Cytometric Stratification of Intracellular Serine Protease Activity in Neutrophils.
Super-silent FRET Sensor Enables Live Cell Imaging and Flow Cytometric Stratification of Intracellular Serine Protease Activity in Neutrophils.
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DOI：
10.1038/s41598-018-31391-9
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发表时间：
2018-09-10
影响因子：
4.6
通讯作者：
Dhaliwal K
中科院分区：
文献类型：
--
作者：
Craven TH;Avlonitis N;McDonald N;Walton T;Scholefield E;Akram AR;Walsh TS;Haslett C;Bradley M;Dhaliwal K
丝氨酸蛋白酶由中性粒细胞释放，主要作为抗菌蛋白，但过度和不平衡的丝氨酸蛋白酶活性会导致严重的宿主组织损伤。本文报道了一种基于多支荧光猝灭剂的新型化学传感器的合成。它是超沉默的，在被典型丝氨酸蛋白酶人中性粒细胞弹性蛋白酶去淬灭之前没有荧光，迅速进入人中性粒细胞，并被丝氨酸蛋白酶抑制剂抑制。该传感器可以实时成像人类中性粒细胞内的细胞内丝氨酸蛋白酶活性，并证明了多价支架与FRET肽的独特组合代表了一种新的有效策略，可以产生超静音传感器，使细胞内蛋白酶可视化，并可能实现中性粒细胞的全血分析。
Serine proteases are released by neutrophils to act primarily as antimicrobial proteins but excessive and unbalanced serine protease activity results in serious host tissue damage. Here the synthesis of a novel chemical sensor based on a multi-branched fluorescence quencher is reported. It is super-silent, exhibiting no fluorescence until de-quenched by the exemplar serine protease human neutrophil elastase, rapidly enters human neutrophils, and is inhibited by serine protease inhibitors. This sensor allows live imaging of intracellular serine protease activity within human neutrophils and demonstrates that the unique combination of a multivalent scaffold combined with a FRET peptide represents a novel and efficient strategy to generate super-silent sensors that permit the visualisation of intracellular proteases and may enable point of care whole blood profiling of neutrophils.
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