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Electrochemical Evaluation of the Mechanism of Acetylcholinesterase Inhibition Based on an Electrodeposited Thin Film

机译:基于电沉积薄膜的乙酰胆碱酯酶抑制机理的电化学评估

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An interface embedded gold nanoparticles in sol-gel thin film was constructed by one-stepelectrochemical deposition. Acetylcholinesterase (AChE) was physically absorbed onto to the interfaceto form a thin enzymatic layer. The proposed thin enzymatic layer, having kinetics similar to that ofthe enzyme in solution, provides an ideal sensing platform to electrochemically evaluate the chemicalmechanism of enzyme inhibition. Lineweaver-Burk plot and surface plasmon resonance confirmed thatthe inhibition of AChE by malathion followed an irreversible mechanism and was a mixed type of2+ 3+competitive and noncompetitive. On the contrary, the degrees of inhibition by Pb and Fe wereindependent of the incubation time and the AChE concentrations, showing the reversibility of theinhibition. Furthermore, UV-vis absorption spectra indicated that the AChE mediated the hydrolysis of3+ 2+ 2+ acetylthiocholine to yield a reducing agent thiocholine that reduced Fe to Fe and Fe presented aneffect of activation. To meet the demand of the biosensor design, we further investigated therelationship between inhibition percentage and both incubation time and inhibitor concentration. Theenzyme sensitivity to solvent effects and reactivation of the biosensor were also evaluated. It isanticipated that a rapid evaluation of the chemical mechanism of AChE inhibition could paves the wayto rationally design biosensors and new compounds, as candidates for the treatment of Alzheimerdisease and pesticides.
机译:通过一步电化学沉积,在溶胶-凝胶薄膜中形成了界面嵌入金纳米颗粒。乙酰胆碱酯酶(AChE)被物理吸收到界面上,形成一个薄的酶层。所提议的薄酶层具有与溶液中的酶相似的动力学,为电化学评估酶抑制的化学机理提供了理想的传感平台。 Lineweaver-Burk图和表面等离子体共振证明,马拉硫磷对AChE的抑制作用是不可逆的,是2+ 3+竞争性和非竞争性的混合类型。相反,铅和铁的抑制程度与孵育时间和AChE浓度无关,显示了抑制作用的可逆性。此外,紫外可见吸收光谱表明,AChE介导3+ 2+ 2+乙酰胆碱的水解,生成还原剂硫胆碱,将Fe还原为Fe,Fe表现出活化作用。为了满足生物传感器设计的需求,我们进一步研究了抑制百分比与孵育时间和抑制剂浓度之间的关系。还评估了酶对溶剂作用的敏感性和生物传感器的活化。可以预见,对AChE抑制作用的化学机理的快速评估可以为合理设计生物传感器和新化合物铺平道路,以作为治疗老年痴呆症和农药的候选药物。

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