Institute of Pesticide Science, College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, China.
Provincial Key Laboratory for Botanical Pesticide R&D of Shaanxi, Yangling, Shaanxi 712100, China.
Toxins (Basel). 2017 Dec 22;10(1):7. doi: 10.3390/toxins10010007.
Periplocoside P (PSP), a novel compound isolated from Bunge, possesses insecticidal activity against some lepidopterans, such as . In the brush border membrane vesicles of the midgut epithelium are the initial site of action of periplocosides. We conducted two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time of flight/time of flight mass spectrometry analysis to analyze differentially expressed proteins (DEPs) from periplocoside P (PSP)-treated . We successfully isolated seven up-regulated and three down-regulated DEPs that have been previously identified, as well as a novel DEP. The DEPs are implicated in protein degradation, transporter, folding, and synthesis, and in juvenile hormone biosynthesis. DEPs involved in the oxidative phosphorylation energy metabolism pathway are enriched. Through real-time polymerase chain reaction assay, we confirmed that expression is significantly up-regulated expression levels in PSP-treated larvae. Enzymology validation further indicated that PSP can significantly inhibit V-type ATPase activity in a concentration-dependent manner. Given these results, we speculate that in , the V-type ATPase A subunit in the midgut epithelium is the putative target binding site of periplocosides. This finding provides preliminary evidence for the mode of action of periplocosides.
杠柳毒苷 P（PSP）是从杠柳中分离得到的一种新型化合物，对鳞翅目昆虫具有杀虫活性，如小菜蛾。在肠上皮的刷状缘膜囊泡中，杠柳毒苷的作用部位是最初的作用部位。我们通过二维凝胶电泳和基质辅助激光解吸/电离飞行时间/飞行时间质谱分析，分析了杠柳毒苷 P（PSP）处理的小菜蛾中差异表达的蛋白质（DEPs）。我们成功分离出 7 个上调和 3 个下调的 DEP，这些 DEP 以前已经被鉴定出来，还有一个新的 DEP。这些 DEP 参与蛋白质降解、转运、折叠和合成以及保幼激素生物合成。参与氧化磷酸化能量代谢途径的 DEP 被富集。通过实时聚合酶链反应测定，我们证实 PSP 处理的小菜蛾幼虫中表达显著上调。酶学验证进一步表明，PSP 可以显著抑制 V 型 ATP 酶活性，呈浓度依赖性。鉴于这些结果，我们推测在小菜蛾中，肠上皮中的 V 型 ATP 酶 A 亚基是杠柳毒苷的假定靶结合位点。这一发现为杠柳毒苷的作用模式提供了初步证据。
