College of Chemistry and Pharmacy, Northwest A&F University, Yangling 712100, China.
College of Life Sciences, Northwest A&F University, Yangling 712100, China.
Toxins (Basel). 2019 Feb 22;11(2):130. doi: 10.3390/toxins11020130.
Celangulin V (CV) is a compound isolated from Max that has a toxic activity against agricultural insect pests. CV can bind to subunits a, H, and B of the vacuolar ATPase (V-ATPase) in the midgut epithelial cells of insects. However, the mechanism of action of CV is still unclear. In this study, the soluble complex of the V-ATPase A subunit mutant TSCA which avoids the feedback inhibition by the hydrolysate ADP and V-ATPase B subunit were obtained and then purified using affinity chromatography. The H⁺K⁺-ATPase activity of the complex and the inhibitory activity of CV on ATP hydrolysis were determined. The results suggest that CV inhibits the ATP hydrolysis, resulting in an insecticidal effect. Additionally, the homology modeling of the AB complex and molecular docking results indicate that CV can competitively bind to the AB complex at the ATP binding site, which inhibits ATP hydrolysis. These findings suggest that the AB subunits complex is one of the potential targets for CV and is important for understanding the mechanism of interaction between CV and V-ATPase.
Celangulin V (CV) 是从 Max 中分离出来的一种化合物，对农业害虫具有毒性活性。CV 可以与昆虫中肠上皮细胞中的 vacuolar ATPase (V-ATPase) 的亚基 a、H 和 B 结合。然而，CV 的作用机制尚不清楚。在这项研究中，获得了避免水解 ADP 反馈抑制的 V-ATPase A 亚基突变体 TSCA 的可溶性复合物，然后使用亲和层析进行纯化。测定了复合物的 H⁺K⁺-ATPase 活性和 CV 对 ATP 水解的抑制活性。结果表明，CV 抑制 ATP 水解，从而产生杀虫作用。此外，AB 复合物的同源建模和分子对接结果表明，CV 可以在 ATP 结合位点与 AB 复合物竞争性结合，从而抑制 ATP 水解。这些发现表明 AB 亚基复合物是 CV 的潜在靶标之一，对于理解 CV 与 V-ATPase 之间的相互作用机制很重要。
