Convenient and continuous fluorometric assay method for acetylcholinesterase and inhibitor screening based on the aggregation-induced emission.
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.
Wang
Ming
M
Beijing National Laboratory for Molecular Sciences, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100080, China.
Gu
Xinggui
X
Zhang
Guanxin
G
Zhang
Deqing
D
Zhu
Daoben
D
D000110:Acetylcholinesterase / Q000032:analysis* / Q000378:metabolism*; D002800:Cholinesterase Inhibitors / Q000032:analysis* / Q000378:metabolism*; D005030:Ethylenes / Q000378:metabolism; D009227:Myristic Acids / Q000378:metabolism; D013050:Spectrometry, Fluorescence / Q000379:methods
D002800:Cholinesterase Inhibitors; D005030:Ethylenes; D009227:Myristic Acids; D000110:Acetylcholinesterase