Agglutination of Human Polyomaviruses by Using a Tetravalent Glycocluster as a Cross-Linker
Agglutination of Human Polyomaviruses by Using a Tetravalent Glycocluster as a Cross-Linker
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DOI：
10.1021/acsomega.0c03269
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发表时间：
2020-09-01
期刊：
影响因子：
4.1
通讯作者：
Suzuki, Tetsuro
中科院分区：
文献类型：
--
作者：
Ogata, Makoto;Onoda, Takashi;Suzuki, Tetsuro
发现两种四价双头唾液酸糖苷能够与默克尔细胞多瘤病毒（MCPyV-LP）的病毒样颗粒结合，所述双头唾液酸糖苷在Neu 5Ac α 2，6 Gal β 1，4GlcNAc单元与乙二醇四乙酸（EGTA）支架之间具有短/长间隔。通过动态光散射（DLS）评估四价配体与MCPyV-LP之间的结合过程和相互作用的时间过程。在向MCPyV-LP添加增加浓度的配体时，形成更大的交联聚集体，直到达到最大尺寸。具有短间隔基的四价配体的结合比具有长间隔基的四价配体的结合更强。在凝集过程中，观察到前一种配体与病毒的结合分两个阶段进行。第一步是自发形成包含交联配体-病毒复合物的小聚集体。在第二步中，通过最初产生的小聚集体之间的合作结合，聚集体逐渐变大。在透射电子显微镜中，观察到所得复合物形成聚集体，其中配体与病毒颗粒紧密堆积。通过简单的膜过滤测定进一步证实了交联的相互作用，其中当与配体复合时，病毒样颗粒保留在膜上。该试验还显示，当与配体复合时，可有效捕获致病性、感染性人多瘤病毒JCPyV的颗粒，表明其可能用作在病毒聚集条件下通过过滤捕获病毒的方法。总的来说，这些结果表明四价糖簇不仅作为凝集MCPyV-LP的配体，而且作为捕获致病性病毒的配体。
Two kinds of tetravalent double-headed sialo-glycosides with short/long spacers between the Neu5Ac alpha 2,6Gal beta 1,4GlcNAc unit and ethylene glycol tetraacetic acid (EGTA) scaffold were found to be capable of binding to virus-like particles of Merkel cell polyomavirus (MCPyV-LP). The binding process and time course of interaction between the tetravalent ligand and MCPyV-LP were assessed by dynamic light scattering (DLS). On the addition of increasing concentrations of ligand to MCPyV-LP, larger cross-linked aggregates formed until a maximum size was reached. The binding was stronger for the tetravalent ligand with a short spacer than for that with a long spacer. The binding of the former ligand to the virus was observed to proceed in two stages during agglutination. The first step was the spontaneous formation of small aggregates comprising the crosslinked ligand-virus complex. In the second step, the aggregates grew successively larger by cooperative binding among the initially produced small aggregates. In transmission electron microscopy, the resulting complex was observed to form aggregates in which the ligands were closely packed with the virus particles. The cross-linked interaction was further confirmed by a simple membrane filtration assay in which the virus-like particles were retained on the membrane when complexed with a ligand. The assay also showed the effective capture of particles of pathogenic, infectious human polyomavirus JCPyV when complexed with a ligand, suggesting its possible application as a method for trapping viruses by filtration under conditions of virus aggregation. Collectively, these results show that the tetravalent glycocluster serves as a ligand not only for agglutinating MCPyV-LP but also for trapping the pathogenic virus.