超静音 FRET 传感器可用于活细胞成像和流式细胞术对中性粒细胞内丝氨酸蛋白酶活性进行分层。
Super-silent FRET Sensor Enables Live Cell Imaging and Flow Cytometric Stratification of Intracellular Serine Protease Activity in Neutrophils.
机构信息
EPSRC Proteus IRC, MRC Centre for Inflammation Research, University of Edinburgh, EH16 4TJ, Edinburgh, United Kingdom.
EaStChem, School of Chemistry, University of Edinburgh, King's Buildings, EH9 3JN, Edinburgh, United Kingdom.
出版信息
Sci Rep. 2018 Sep 10;8(1):13490. doi: 10.1038/s41598-018-31391-9.
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.
丝氨酸蛋白酶由中性粒细胞释放,主要作为抗菌蛋白发挥作用,但过度和失衡的丝氨酸蛋白酶活性会导致严重的宿主组织损伤。在此,我们报道了一种基于多分支荧光猝灭剂的新型化学传感器的合成。它是超静默的,在被典型的丝氨酸蛋白酶人中性粒细胞弹性蛋白酶去猝灭之前没有荧光,它可以快速进入人中性粒细胞,并被丝氨酸蛋白酶抑制剂抑制。该传感器允许在人中性粒细胞内对细胞内丝氨酸蛋白酶活性进行活细胞成像,并证明多价支架与 FRET 肽的独特组合代表了一种新颖有效的策略,可以生成超静默的传感器,从而可以可视化细胞内蛋白酶,并可能实现即时护理全血中中性粒细胞的分析。
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