华南农业大学学报2024，Vol.45Issue(4)：542-549,8.DOI:10.7671/j.issn.1001-411X.202310027
荷包牡丹碱对家蝇GABA受体作用机理探究
Study on the mechanism of action of bicuculline on GABA receptor in Musca domestica
摘要
Abstract
[Objective]In order to explore the inhibitory mechanism of bicuculline on GABA receptor,which was an important target of insecticide.[Method]Based on alternately splicing at exon 3(a or b)and exon 6(c or d),Musca domestica GABA receptor subunit gene MdRdl yields ac,ad,bc and bd spliceosomes.Two-electrode voltage clamp technique was used to study the inhibitory effect of bicuculline on the ion current of four spliceosomes of MdRdl,characterizing whether its sensitivity was related to alternate splicing of the exons.The molecular docking technique was used to analyze the possible binding sites of bicuculline and GABA receptor of M.domestica.The interaction effect between bicuculline and the allosteric inhibitor of GABA receptors,broflanilide,was verified using the co-stimulation method.[Result]The IC50s of bicuculline on the four spliceosomes MdRdlac,MdRdlbc,MdRdlad and MdRdlbd were 1.274,0.948,4.702 and 3.698 mmol/L,on their I274F mutants were 15.46,21.67,42.83 and 52.33 mmol/L,respectively.The IC50s of four mutants increased by about 11,22,8 and 13 times compared to their wildtype spliceosomes,respectively.Bicuculline and broflanilide showed interactions with amino acid residues encoded by the exon 6 of the GABA receptor.[Conclusion]The isoleucine residue at position 274 on the GABA receptor was the key target for the interaction between bicuculline and GABA receptor of M.domestica.This isoleucine residue and the amino acid residues encoded by exon 6 together are potentially to be developed as the synergistic allosteric inhibitors of insect GABA receptors.关键词
荷包牡丹碱/γ-氨基丁酸受体/协同增效/分子对接Key words
Bicuculline/γ-aminobutyric acid receptor/Synergistic interaction/Molecular docking分类
农业科技引用本文复制引用
刘夏娜,江定心..荷包牡丹碱对家蝇GABA受体作用机理探究[J].华南农业大学学报,2024,45(4):542-549,8.基金项目
国家自然科学基金(32372576) （32372576）
广东省基础与应用基础研究基金(2021A1515011277) （2021A1515011277）
