Biomedical Research Institute, Central 6, and ‡Nanoelectronics Research Centre, Central 4, National Institute of Advanced Industrial Science and Technology , 1-1-1 Higashi, Tsukuba City 305-8566, Ibaraki, Japan.
Anal Chem. 2013 Nov 5;85(21):10455-62. doi: 10.1021/ac4025522. Epub 2013 Oct 15.
The entry of herpes simplex virus into host cells involves a complex series of events that require concerted inputs from multiple HSV glycoproteins. Among these glycoproteins, the gD protein of HSV-1 and HSV-2 plays an important role for host receptor binding and membrane fusion. In the present study, we evaluated the ability of different sulfated saccharides to interfere with gD-host receptor (HVEM) interactions using our recently reported molecular assay (Gopinath, S. C. B.; Hayashi, K.; Kumar, P. K. R. J. Virol. 2012, 86, 6732-6744). Initially, we tested the ability of heparan sulfate to interfere with the HVEM-HSV-1 gD interaction and found that heparan sulfate is able to interfere efficiently, with an apparent EC50 of 2.1 μM. In addition, we tested different synthetic sulfated polysaccharides and natural sulfated polysaccharides from an edible alga, Sargassum horneri , after fractionation into different sizes and sulfate and uronic acid contents. Six polysaccharides isolated from S. horneri were found to efficiently interfere with the HVEM-gD interaction. Three others caused moderate interference, and five caused weak interference. These results were confirmed with plaque assays, and good agreement was found with the results of the SPR assay for the identification of compounds that interfere with HVEM-HSV-1 gD binding. These studies suggest that our molecular assay based on surface plasmon resonance is not only useful for the analysis of viral-host protein interactions but is also applicable for the routine screening of compounds to identify those that interfere with the first step of viral entry, thus facilitating the rapid development of novel antiviral compounds that target HSV.
单纯疱疹病毒进入宿主细胞涉及一系列复杂的事件，需要多种单纯疱疹病毒糖蛋白的协同输入。在这些糖蛋白中，HSV-1 和 HSV-2 的 gD 蛋白在宿主受体结合和膜融合中起重要作用。在本研究中，我们使用最近报道的分子测定法（Gopinath，S.C.B.；Hayashi，K.；Kumar，P.K.R. J. Virol. 2012, 86, 6732-6744）评估了不同硫酸化糖对 gD-宿主受体（HVEM）相互作用的干扰能力。最初，我们测试了硫酸乙酰肝素对 HVEM-HSV-1 gD 相互作用的干扰能力，发现硫酸乙酰肝素能够有效地干扰，表观 EC50 为 2.1 μM。此外，我们测试了不同的合成硫酸化多糖和天然硫酸化多糖，这些多糖来自一种可食用海藻，马尾藻，经过不同大小和硫酸和糖醛酸含量的分离。从马尾藻中分离出的六种多糖被发现能有效地干扰 HVEM-gD 相互作用。另外三种引起中度干扰，五种引起弱干扰。这些结果通过蚀斑试验得到了证实，并且与 SPR 测定法鉴定干扰 HVEM-HSV-1 gD 结合的化合物的结果吻合良好。这些研究表明，我们基于表面等离子体共振的分子测定法不仅可用于分析病毒-宿主蛋白相互作用，还可用于常规筛选化合物，以鉴定那些干扰病毒进入第一步的化合物，从而促进针对单纯疱疹病毒的新型抗病毒化合物的快速开发。
