An electrochemical platform for acetylcholinesterase activity assay and inhibitors screening based on Michael addition reaction between thiocholine and catechol-terminated SAMs.
An electrochemical platform for acetylcholinesterase activity assay and inhibitors screening based on Michael addition reaction between thiocholine and catechol-terminated SAMs.
复制标题
DOI：
10.1039/c1an15601b
复制
发表时间：
2011-11
期刊：
影响因子：
--
通讯作者：
Yuan Tian;S. Ye;Xinhao Shi;Li Jing;Cong Liang;Y. Xian
中科院分区：
文献类型：
--
作者：
Yuan Tian;S. Ye;Xinhao Shi;Li Jing;Cong Liang;Y. Xian
基于乙酰胆碱酯酶（AChE）水解产物硫代胆碱（AsCh）与邻苯二酚封端的自组装膜（SAMs）在金电极上的Michael加成反应，建立了一种用于AChE活性测定及其抑制剂筛选的电化学平台.为了进一步理解和证实反应机理，研究了半胱氨酸（CYS）和谷胱甘肽（GSH）两种模型化合物与邻苯二酚封端的自组装膜的Michael加成反应的电化学行为。采用安培法研究了三种乙酰胆碱酯酶抑制剂（他克林、克百威和甲基对硫磷）的酶动力学和抑制作用。在这三种抑制剂中，他克林表现出最强的抑制作用，这是由每个抑制剂对酶活性的影响的动力学研究的结果数据得到加强。
An electrochemical platform for acetylcholinesterase (AChE) activity assay and its inhibitors screening is developed based on the Michael addition reaction of thiocholine, the hydrolysis product of acetylthiocholine (AsCh) in the presence of AChE, with the electrogenerated o-quinone of catechol-terminated SAMs on a gold electrode. For understanding and confirming the mechanism of the reaction, the electrochemical behaviors of Michael addition reaction of two model compounds, cysteine (CYS) and glutathione (GSH), towards the catechol-terminated SAMs have been studied. The enzyme kinetics and the inhibition effects of three types of AChE inhibitors, which are tacrine, carbofuran and parathion-methyl, have been investigated using an amperometric method. Among these three inhibitors, tacrine exhibits the strongest inhibiting effect, which is reinforced by the resulting data of kinetic studies on each inhibitor's influence upon the enzyme activity.