The expression of soluble functional alpha 7-nicotinic acetylcholine receptors in E. coli and its high-affinity binding to neonicotinoid pesticides

文献类型： 外文期刊

第一作者： Zhou, Qiaoling

作者： Zhou, Qiaoling;Yan, Ru;Chen, Mengli;Xiang, Dandan;Xu, Xiaojun;Zhou, Qiaoling;Yan, Ru;Chen, Mengli;Guo, Yirong;Zhu, Guonian;Xiang, Dandan;Xu, Xiaojun

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关键词： Nicotinic acetylcholine receptor; alpha 7 subunit; Neonicotinoids; E. coli; SPR

期刊名称：PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY （ 影响因子：3.963； 五年影响因子：4.454 ）

ISSN： 0048-3575

年卷期： 2020 年 164 卷

页码：

收录情况： SCI

摘要： Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels mediating fast cholinergic synaptic transmission in nervous system. In insects, nAChRs are the target sites for several naturally occurring and synthetic compounds, including the neonicotinoid insecticides. So far, one of the major strategies to explore the interaction of nAChR and ligands is based on the heterologous expression of nAChR, which is tough, and needs to be explored. In this study, we expressed and purified extracellular domain of rat alpha 7 subunit (R alpha 7-ECD), the binding site of the ligands in E. coli and determined the interactions and kinetic constants of neonicotinoid insecticides with R alpha 7-ECD. The recombinant R alpha 7-ECD is water-soluble and appears to be correctly folded. The interactions of three neonicotinoid pesticides with R alpha 7-ECD were assessed by surface plasmon resonance (SPR) biosensor. The results revealed a fast association and fast disassociation binding mode of R alpha 7-ECD/pesticides complexes with the KD value of clothianidin (6.414E-9 M) > imidacloprid (9.030E-9 M) > acetamiprid (2.874E-6 M), respectively. This study demonstrated that the nAChR expressed from E. coli was functional, and SPR biosensor technology would be a good alternative for characterizing members of nAChR receptor family.

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