Edmonds Kearstin, Dutch Rebecca
Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, KY, USA.
Methods Mol Biol. 2025;2948:73-83. doi: 10.1007/978-1-0716-4666-3_5.
The methods detailed here measure the membrane fusion activity of viral fusion proteins and are optimized for the respiratory syncytial virus (RSV). RSV utilizes a class I fusion protein (F) to facilitate fusion between a viral and cellular membrane to allow viral entry into host cells. This process is carried out with the assistance of the glycoprotein (G), which attaches to the host-cell surface, allowing F to interact with the target cell. The F protein then undergoes conformational changes, which drive membrane fusion, thus creating a path for viral entry. The fusogenic activity of viral fusion proteins can be quantified by observing and measuring syncytia formation via wide-field microscopy. Syncytia are multinucleated cell masses formed by cell-cell membrane fusion, which in this case are driven by viral fusion proteins expressed on the host-cell surface.
此处详述的方法用于测量病毒融合蛋白的膜融合活性，并且针对呼吸道合胞病毒（RSV）进行了优化。RSV利用I类融合蛋白（F）促进病毒膜与细胞膜之间的融合，以使病毒进入宿主细胞。此过程在糖蛋白（G）的协助下进行，糖蛋白附着于宿主细胞表面，使F能够与靶细胞相互作用。然后F蛋白发生构象变化，驱动膜融合，从而为病毒进入创造一条途径。病毒融合蛋白的融合活性可以通过宽场显微镜观察和测量多核巨细胞形成来定量。多核巨细胞是由细胞-细胞膜融合形成的多核细胞团，在这种情况下是由宿主细胞表面表达的病毒融合蛋白驱动的。
