Department of Chemical Biology & Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, The Netherlands.
Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, California, USA.
Protein Sci. 2024 Apr;33(4):e4974. doi: 10.1002/pro.4974.
Enveloped viruses carry one or multiple proteins with receptor-binding functionalities. Functional receptors can be glycans, proteinaceous, or both; therefore, recombinant protein approaches are instrumental in attaining new insights regarding viral envelope protein receptor-binding properties. Visualizing and measuring receptor binding typically entails antibody detection or direct labeling, whereas direct fluorescent fusions are attractive tools in molecular biology. Here, we report a suite of distinct fluorescent fusions, both N- and C-terminal, for influenza A virus hemagglutinins and SARS-CoV-2 spike RBD. The proteins contained three or six fluorescent protein barrels and were applied directly to cells to assess receptor binding properties.
包膜病毒携带一种或多种具有受体结合功能的蛋白质。功能性受体可以是糖基、蛋白质或两者兼有；因此，重组蛋白方法对于深入了解病毒包膜蛋白的受体结合特性非常重要。可视化和测量受体结合通常需要抗体检测或直接标记，而直接荧光融合则是分子生物学中的一种有吸引力的工具。在这里，我们报告了一系列不同的荧光融合物，包括流感 A 病毒血凝素和 SARS-CoV-2 刺突 RBD 的 N 端和 C 端。这些蛋白包含三或六个荧光蛋白桶，并直接应用于细胞，以评估受体结合特性。
