基于聚集诱导发光的乙酰胆碱酯酶及抑制剂筛选的便捷连续荧光检测法

Convenient and continuous fluorometric assay method for acetylcholinesterase and inhibitor screening based on the aggregation-induced emission.

作者信息

Wang Ming, Gu Xinggui, Zhang Guanxin, Zhang Deqing, Zhu Daoben

机构信息

Beijing National Laboratory for Molecular Sciences, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100080, China.

出版信息

Anal Chem. 2009 Jun 1;81(11):4444-9. doi: 10.1021/ac9002722.

DOI:10.1021/ac9002722

PMID:19374428

Abstract

A new convenient and continuous fluorometric assay method for acetylcholinesterase (AChE) and its inhibitor screening is successfully established with the ensemble of 1 [a TPE (tetraphenylethylene) compound with two sulfonate (-SO(3)(-)) units] and myristoylcholine (an amphiphilic compound as a good substrate of AChE). This new assay method is designed by making use of the aggregation-induced emission (AIE) feature of TPE compounds. Both dynamic light scattering (DLS) and fluorescence confocal microscopic measurements indicated the formation of aggregation complex for the ensemble of 1 and myristoylcholine and further disassembly of the aggregation complex after introducing AChE. The analysis for AChE can be carried out continuously, and AChE with concentration as low as 0.5 U/mL can be assayed. The results also clearly demonstrate the usefulness of this convenient assay method for kinetic study of AChE-catalyzed myristoylcholine hydrolysis and screening inhibitors of AChE. Given its simplicity and easy operation, this method may extend to high-throughput screening of AChE inhibitors and relevant Alzheimer's disease (AD) drug discovery.

摘要

利用1（一种带有两个磺酸根（-SO(3)(-)）单元的四苯乙烯（TPE）化合物）和肉豆蔻酰胆碱（一种两亲性化合物，作为乙酰胆碱酯酶（AChE）的良好底物）的组合，成功建立了一种用于AChE及其抑制剂筛选的便捷连续荧光测定方法。这种新的测定方法利用了TPE化合物的聚集诱导发光（AIE）特性。动态光散射（DLS）和荧光共聚焦显微镜测量均表明，1与肉豆蔻酰胆碱的组合形成了聚集复合物，引入AChE后聚集复合物进一步解体。对AChE的分析可以连续进行，低至0.5 U/mL的AChE浓度也可被测定。结果还清楚地证明了这种便捷测定方法在AChE催化肉豆蔻酰胆碱水解的动力学研究和AChE抑制剂筛选中的实用性。鉴于其简单性和易操作性，该方法可能扩展到AChE抑制剂的高通量筛选及相关阿尔茨海默病（AD）药物的发现。