分子靶标导向的新农药创制是新世纪农药科学研究与发展的前沿。在已完成的国家基金项目研究中，发现杀虫植物杠柳（Periploca sepium）的活性成分杠柳毒素periplocoside P（PSP）的靶蛋白为昆虫中肠刷状缘膜囊泡上V-ATP酶A亚基，但V-ATP酶A亚基与PSP的结合位点并不清楚。基于已构建的V-ATP酶A亚基真核表达系统pBacPAK9/VatpA，本项申请拟运用分子对接、分子动力学和多尺度组合等方法确定PSP与V-ATP酶A亚基结合的3D结合模型，预测PSP结合位点并进行关键氨基酸定点突变，验证和明确PSP靶标结合位点，阐明互作机理；应用分子对接虚拟筛选—蛋白亲和筛选—离体/活体生测进行对靶高活性先导化合物筛选，探究V-ATP酶A亚基作为杀虫剂靶标的成药性，为进一步创制作用于昆虫消化系统的新型杀虫剂提供新的靶标分子模型。

The creation of new pesticides guided by molecular targets is a frontier subject in the research and development of pesticide science. In the finished National Fund Projects, we found that the target protein of insecticidal component periplocoside P (PSP) from Periploca sepium is the vacuolar-type H+-ATPase (V-ATPase) A subunit on the brush border membrane vesicles in insect midgut, but the binding sites of PSP remains unclear. Based on the eukaryotic expression system of V-ATPase A subunit, pBacPAK9/VatpA, this project aims to construct the 3D model of V-ATPase A subunit binding with PSP by molecular docking, molecular dynamics and multi-scale combination. The key amino acid of PSP binding sites predicted will be verified by site-directed mutagenesis for elucidating their molecular interaction mechanism. The druggability of V-ATPase A subunit as a insecticidal target will be investigated by means of virtual screening, protein affinity chromatography screening and in vitro/in vivo bioassay for screening high activity lead compound A new target molecular model will be provide for the creation of new insecticides acting on insect digestive system.