A highly selective fluorescent ESIPT probe for the detection of Human carboxylesterase 2 and its biological applications
A highly selective fluorescent ESIPT probe for the detection of Human carboxylesterase 2 and its biological applications
复制标题
一种高选择性荧光ESIPT探针检测人羧酸酯酶2及其生物学应用
DOI：
10.1016/j.bios.2014.10.002
复制
发表时间：
2015-03-15
影响因子：
12.6
通讯作者：
Yang, Ling
中科院分区：
文献类型：
--
作者：
Feng, Lei;Liu, Zhao-Ming;Yang, Ling
以3-羟基黄酮（3-HF）为衍生物，合成了一种新的比率荧光探针，用于人羧酸酯酶2（CE 2）的选择性和灵敏性检测。该探针通过引入4-乙基苯甲酰氧基来调节3-HF的激发态分子内质子转移（ESIPT）发射而设计。在生理条件下，探针1具有良好的稳定性，背景信号很低，但可被CE 2选择性水解，释放出游离的3-HF，使荧光光谱发生显著变化。反应表型分析和化学抑制试验表明，探针1是高度选择性的CE 2超过其他人类水解酶，包括羧酸酯酶1，胆碱酯酶和对氧磷酶。探针1已成功地应用于人体生物样品中CE 2的真实的活性测定，以及以组织制备物为酶源筛选CE 2抑制剂。另外，探针1是细胞膜可渗透的，并且可用于活细胞中内源性CE 2的细胞成像。所有这些特征使得它有可能作为一个有前途的工具，探索个体差异的生物学功能的CE 2，以及快速筛选选择性和有效的抑制剂CE 2的进一步临床使用。(C)2014爱思唯尔有限公司版权所有。
A new ratiometric florescence probe derived from 3-hydroxyflavone (3-HF) has been developed for selective and sensitive detection of human carboxylesterase 2 (CE2). The probe is designed by modulating the excited state intramolecular proton transfer (ESIPT) emission of 3-HF via introducing of 4-ethylbenzoyloxy group. Under physiological conditions, probe 1 displays satisfying stability with very low background signal, but it can be selectively hydrolyzed by CE2 to release free 3-HF which brings remarkable changes in fluorescence spectrum. Both reaction phenotyping and chemical inhibition assays demonstrate that probe 1 is highly selective for CE2 over other human hydrolases including carboxylesterase 1, cholinesterases and paraoxonases. Probe 1 has been applied successfully to measure the real activities of CE2 in human biological samples, as well as to screen CE2 inhibitors by using tissue preparations as the enzymes sources. Additionally, probe 1 is cell membrane permeable and can be used for cellular imaging of endogenous CE2 in living cells. All of these features make it possible to serve as a promising tool for exploring the individual differences in biological function of CE2, as well as for rapid screening of selective and potent inhibitors of CE2 for further clinical use. (C) 2014 Elsevier B.V. All rights reserved.