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一种绵铃醇电化学受体传感器及其动力学。

A bombykol electrochemical receptor sensor and its kinetics.

机构信息

College of Biotechnology & Food Science, Tianjin University of Commerce, Tianjin 300134, China; Tianjin Key Laboratory of Food Biotechnology, Tianjin 300134, China.

College of Biotechnology & Food Science, Tianjin University of Commerce, Tianjin 300134, China.

出版信息

Bioelectrochemistry. 2019 Aug;128:263-273. doi: 10.1016/j.bioelechem.2019.04.009. Epub 2019 Apr 23.

Abstract

This study aimed to explore the interaction between bombykol and BmOR1 and also provide a paradigm for agroforestry pest control. The electrochemical biosensor signal amplification system was used: nanogold with horseradish peroxidase. An electrochemical bilayer nanogold membrane receptor sensor was developed using the following schemes and processes: twice self-assembly of nanogold and succeeding absorption of Bombyx mori olfactory receptor 1 (BmOR1); sex pheromone-binding protein; spectral scanning and transmission electron microscope to characterize nanogold sol; and atomic force microscope, cyclic voltammetry, and AC impedance methods to characterize individual processes of sensor assembly. The amperometric I-T curve was adopted to measure the response current upon interaction with different concentrations of bombykol (diluted in phosphate-buffered saline) and BmOR1. The results demonstrated the receptor-ligand interaction pattern, which was similar to enzymatic reaction kinetics, with the activation constant Ka of up to 8.57 × 10 mol/L and signal magnification of about 10,000-fold. In this study, the simulation of intracellular receptor signaling cascade by an electrochemical signal amplification system helped in directly measuring BmOR1-bombykol ligand interaction and exploring the kinetics after the self-assembly of BmOR1 on the biosensor. It provided a novel platform for future studies on receptor-ligand interaction.

摘要

本研究旨在探讨 Bombykol 与 BmOR1 的相互作用,并为农林害虫防治提供范例。采用电化学生物传感器信号放大系统:辣根过氧化物酶纳米金。采用以下方案和工艺开发电化学双层纳米金膜受体传感器:两次纳米金自组装,随后吸收家蚕嗅觉受体 1(BmOR1);性信息素结合蛋白;纳米金溶胶的光谱扫描和透射电子显微镜表征;原子力显微镜、循环伏安法和交流阻抗法对传感器组装的各个过程进行表征。采用安培电流 I-T 曲线测量与不同浓度 Bombykol(用磷酸盐缓冲液稀释)和 BmOR1 相互作用时的响应电流。结果表明,受体-配体相互作用模式类似于酶促反应动力学,激活常数 Ka 高达 8.57×10-6mol/L,信号放大倍数约为 10000 倍。在这项研究中,通过电化学信号放大系统模拟细胞内受体信号级联,有助于直接测量 BmOR1-bombykol 配体相互作用,并探索 BmOR1 在生物传感器上自组装后的动力学。它为未来的受体-配体相互作用研究提供了一个新的平台。

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