Convenient and Continuous Fluorometric Assay Method for Acetylcholinesterase and Inhibitor Screening Based on the Aggregation-Induced Emission
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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.
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