Faculty of Pharmacy, Osaka Ohtani University, Tondabayashi, Japan.
Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo, Japan.
Methods Mol Biol. 2020;2132:585-595. doi: 10.1007/978-1-0716-0430-4_50.
Rotaviruses are the major etiologic agents of acute gastroenteritis. Viral attachment to the cell surface is crucial to initiate infection. The VP8 domain, the trypsinized cleavage fragment of the outermost spike protein VP4 of rotavirus, has a galectin-like structure required for binding to the cell surface. We used the evanescent-field fluorescence-assisted assay to understand the complex mechanism underlying the virus-glycan/glycoprotein interaction. Besides, we have described virus infection assays, neutralization assay, and pretreatment assay, using cell culture. These approaches using rotavirus particles will provide novel information that has been difficult to obtain from glycan microarray using recombinant VP8.
轮状病毒是急性肠胃炎的主要病原体。病毒与细胞表面的附着对于引发感染至关重要。VP8 结构域是轮状病毒最外层刺突蛋白 VP4 的胰蛋白酶切割片段，具有与细胞表面结合所必需的半乳糖凝集素样结构。我们使用渐逝场荧光辅助测定法来理解病毒-聚糖/糖蛋白相互作用的复杂机制。此外，我们还描述了使用细胞培养进行的病毒感染测定、中和测定和预处理测定。这些使用轮状病毒颗粒的方法将提供新的信息，这些信息很难从使用重组 VP8 的聚糖微阵列中获得。
