Dan Katsuaki, Yamase Toshihiro
Center for Integrated Medical Research, Central Research Laboratory, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160, Japan.
Biomed Pharmacother. 2006 May;60(4):169-73. doi: 10.1016/j.biopha.2006.02.005. Epub 2006 Mar 29.
One of the Keggin-type heteropolyoxotungstates (K7[PTi2W10O40]6H2O:PM-19) is a potent inhibitor of the replication of herpes simplex virus (HSV) both standard strain 169 and the thymidine kinase-defective strain YS-4C-1 in vitro and in vivo. HSV envelope protein, gD, is necessary for virus entry into the cells. Some cellular molecules, such as HVEM, were reported to act as cofactors during the viral entry step. We determined whether PM-19 prevents these interactions between HSV-gD and HVEM. These activities were investigated using the Ciphergen and BIACORE system. Using a protein chip, many kinds of gD-specific binding proteins were captured, but these proteins could not be identified. Several proteins in these gD-binding proteins were inhibited its interaction with gD due to the presence of PM-19. Using the BIACORE system, the affinity of PM-19 to gD was low, because PM-19 has no direct inactivation activity against the virion. The specific binding of HVEM to the gD was shown as KD of 1.1e-9. The affinity of PM-19 for HVEM was high (KD:2e-9). To determine the competitive binding, the PM-19 (10 microg/ml) and several concentrations of HVEM solution mixtures were injected over the gD-fixed sensor surface. Each binding signal was stable in the range of approximately 270-300 RU. In the case of the addition of PM-19 to HVEM solution, the binding signals were elevated by PM-19 dose dependently. These results suggest that the bindings of PM-19 to gD are not disturbed by the presence of HVEM. PM-19 prevents the interaction between HVEM and gD.
Keggin型杂多钨酸盐之一（K7[PTi2W10O40]·6H2O：PM - 19）在体外和体内对单纯疱疹病毒（HSV）的标准株169和胸苷激酶缺陷株YS - 4C - 1的复制均具有强效抑制作用。HSV包膜蛋白gD是病毒进入细胞所必需的。据报道，一些细胞分子，如疱疹病毒侵入介导因子（HVEM），在病毒进入步骤中作为辅助因子发挥作用。我们确定了PM - 19是否能阻止HSV - gD与HVEM之间的这些相互作用。使用Ciphergen和BIACORE系统对这些活性进行了研究。利用蛋白质芯片捕获了多种gD特异性结合蛋白，但这些蛋白无法被鉴定。由于PM - 19的存在，这些gD结合蛋白中的几种蛋白与gD的相互作用受到抑制。使用BIACORE系统，PM - 19与gD的亲和力较低，因为PM - 19对病毒粒子没有直接的灭活活性。HVEM与gD的特异性结合表现为解离常数（KD）为1.1×10⁻⁹。PM - 19对HVEM的亲和力较高（KD：2×10⁻⁹）。为了确定竞争性结合，将PM - 19（10微克/毫升）与几种浓度的HVEM溶液混合物注射到固定有gD的传感器表面。每个结合信号在大约270 - 300响应单位（RU）范围内稳定。在向HVEM溶液中添加PM - 19的情况下，结合信号随PM - 19剂量依赖性升高。这些结果表明，PM - 19与gD的结合不受HVEM存在的干扰。PM - 19可阻止HVEM与gD之间的相互作用。
