Probing Single Virus Binding Sites on Living Mammalian Cells Using AFM
Probing Single Virus Binding Sites on Living Mammalian Cells Using AFM
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DOI：
10.1007/978-1-4939-8591-3_29
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发表时间：
2018-01-01
期刊：
影响因子：
--
通讯作者：
Alsteens, David
中科院分区：
文献类型：
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作者：
Delguste, Martin;Koehler, Melanie;Alsteens, David
在过去的几年中，基于原子力显微镜（AFM）的方法已经发展成为一种强大的多参数工具，可以探测从单一受体到膜和组织的生物样本。如今，基于力-距离曲线的 AFM（基于 FD 的 AFM）能够以高分辨率对活细胞进行成像，并同时定位和表征特定的配体-受体结合事件。在本章中，我们将介绍基于 FD 的 AFM 如何研究病毒与活哺乳动物细胞的结合，并量化描述单个病毒与受体介导的结合的自由能景观的动力学和热力学参数。使用模型病毒，我们探测了与表达其同源受体的细胞的特异性相互作用，并测量了相互作用的亲和力。此外，我们观察到病毒迅速建立了特定的多价相互作用，并发现按顺序形成的每个键都会加强病毒与细胞的附着。
In the last years, atomic force microscopy (AFM)-based approaches have evolved into a powerful multi-parametric tool that allows biological samples ranging from single receptors to membranes and tissues to be probed. Force-distance curve-based AFM (FD-based AFM) nowadays enables to image living cells at high resolution and simultaneously localize and characterize specific ligand-receptor binding events. In this chapter, we present how FD-based AFM permits to investigate virus binding to living mammalian cells and quantify the kinetic and thermodynamic parameters that describe the free-energy landscape of the single virus-receptor-mediated binding. Using a model virus, we probed the specific interaction with cells expressing its cognate receptor and measured the affinity of the interaction. Furthermore, we observed that the virus rapidly established specific multivalent interactions and found that each bond formed in sequence strengthens the attachment of the virus to the cell.