State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University, Guiyang 550025, PR China.
Department of Plant Pathology, College of Agriculture, Guizhou University, Guiyang 550025, PR China.
Pestic Biochem Physiol. 2020 Jun;166:104449. doi: 10.1016/j.pestbp.2019.07.017. Epub 2019 Nov 4.
Tobacco mosaic virus helicase (TMV-Hel) plays important roles in viral multiplication. TMV-Hel is a potential target of anti-TMV agents. Our previous studies expressed and purified TMV-Hel as target protein for cytosinpeptidemycin. In this study, we preform molecular docking to study the binding sites of commercial antiviral agents with TMV-Hel. Then we verify the interactions between the potential anti-TMV agents and TMV-Hel in vitro using Microscale Thermophoresis experiment and study the inhibiting expression of TMV-Hel with the potential anti-TMV agents in vivo using Western-blot (WB) method. The results showed that ribavirin bound to TMV-Hel with a dissociation constant of 1.55 μM by direct interaction with eight binding sites, which was consistent with the docking studies. Ribavirin inhibited the expression of TMV-Hel in Nicotiana benthamiana. Docking studies combined Microscale Thermophoresis and WB experiment provided a new method to screen anti-TMV agents targeting TMV-Hel.
烟草花叶病毒解旋酶（TMV-Hel）在病毒复制中发挥重要作用。TMV-Hel 是抗 TMV 药物的潜在靶点。我们之前的研究将 TMV-Hel 作为胞嘧啶肽霉素的靶蛋白进行了表达和纯化。在这项研究中，我们进行了分子对接，以研究商业抗病毒药物与 TMV-Hel 的结合位点。然后，我们使用微量热泳动实验在体外验证潜在抗 TMV 药物与 TMV-Hel 之间的相互作用，并使用 Western-blot（WB）方法研究潜在抗 TMV 药物在体内对 TMV-Hel 表达的抑制作用。结果表明，利巴韦林通过与八个结合位点的直接相互作用与 TMV-Hel 结合，解离常数为 1.55 μM，这与对接研究结果一致。利巴韦林抑制了 Nicotiana benthamiana 中 TMV-Hel 的表达。对接研究结合微量热泳动和 WB 实验为筛选靶向 TMV-Hel 的抗 TMV 药物提供了一种新方法。
