An electrochemical device for the assay of the interaction between a dioxin receptor and its various ligands
An electrochemical device for the assay of the interaction between a dioxin receptor and its various ligands
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DOI：
10.1016/j.bmcl.2003.10.002
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发表时间：
2004-01-05
影响因子：
2.7
通讯作者：
Katayama, Y
中科院分区：
文献类型：
--
作者：
Murata, M;Gonda, H;Katayama, Y
芳烃受体（Aryl hydrocarbon receptor, AhR）是一种介导多种化学物质毒性和生物效应的配体激活转录因子。虽然关于平面芳香烃AhR配体的大量信息是可用的，但关于可以结合和激活AhR的化学结构的实际光谱的信息是不足的。为了确定化学物质与人体AhR （hAhR）的结合亲和力，我们构建了一个在电极表面携带hAhR配体结合域的电化学体系。利用T7表达系统在大肠杆菌中表达重组hAhR配体结合域，将其固定在金电极上。这种基于配体-受体相互作用的生物传感器的特异性与其他体外筛选方法相当。受体修饰电极可以快速检测配体与hAhR的结合。电化学测量可在5分钟内完成。该电化学筛选系统快速，成本低，适用于高通量应用，而不会牺牲灵敏度或选择性。(C) 2003 Elsevier Ltd.版权所有。
Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that mediates the toxic and biological effects of a variety of chemicals. Although a significant amount of information is available with respect to the planar aromatic hydrocarbon AhR ligands, information on the actual spectrum of chemical structures that can bind to and activate the AhR is insufficient. In order to determine the binding affinities of chemicals to the human AhR (hAhR), we constructed an electrochemical system which carries the hAhR ligand-binding domain on the electrode surface. The recombinant hAhR ligand-binding domain that was expressed in Escherichia coli using a T7 expression system was immobilized on a gold electrode. The specificity of this biosensor based on a ligand-receptor interaction was comparable to other in vitro screening methods. The receptor-modified electrode can rapidly detect the binding of ligands to hAhR. The electrochemical measurement can be carried out within just 5 min. This electrochemical screening system is rapid, low in cost, and adaptable to high-throughput applications without sacrificing either sensitivity or selectivity. (C) 2003 Elsevier Ltd. All rights reserved.