Department of Microbiology, College of Life Sciences, Nankai University, Tianjin 300071, China.
State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, China.
Toxins (Basel). 2018 Dec 18;10(12):546. doi: 10.3390/toxins10120546.
Vegetative insecticidal proteins (Vips), which are secreted by some strains during vegetative growth, exhibit high virulence to many pests. Vip3A proteins have been used commercially both in some bio-insecticides and in transgenic crops; however, compared with insecticidal crystal proteins, the mechanism of action of Vip3A is still unclear. In this work, we indicated that the fibroblast growth factor receptor-like protein (Sf-FGFR) from the membrane of Sf9 cells could bind to Vip3Aa. The interaction between Vip3Aa and Sf-FGFR was confirmed by pull-down assays and dot blotting experiment in vitro. The binding affinity between Vip3Aa and extracellular regions of Sf-FGFR (GST-FGFR-N) was determined by microscale thermophoresis assay (MST). Moreover, Vip3Aa-Flag could be co-immunoprecipitated with Sf-FGFR-V5 ex vivo. Furthermore, knockdown of gene in Sf9 cells resulted in reducing the mortality of those cells to Vip3Aa. In summary, our data indicated that Sf-FGFR is a novel receptor for Vip3Aa.
植物源杀虫蛋白(Vips)在营养生长阶段由一些菌株分泌,对许多害虫具有高毒性。Vip3A 蛋白已被用于一些生物杀虫剂和转基因作物中;然而,与杀虫晶体蛋白相比,Vip3A 的作用机制仍不清楚。在这项工作中,我们表明 Sf9 细胞膜上的成纤维细胞生长因子受体样蛋白(Sf-FGFR)可以与 Vip3Aa 结合。通过体外下拉实验和斑点印迹实验证实了 Vip3Aa 与 Sf-FGFR 之间的相互作用。通过微量热泳动分析(MST)测定了 Vip3Aa 与 Sf-FGFR 胞外区(GST-FGFR-N)之间的结合亲和力。此外,Vip3Aa-Flag 可以与 Sf-FGFR-V5 在细胞外共沉淀。此外,Sf9 细胞中基因的敲低导致细胞对 Vip3Aa 的死亡率降低。总之,我们的数据表明 Sf-FGFR 是 Vip3Aa 的一个新型受体。