Viral envelope proteins fused to multiple distinct fluorescent reporters to probe receptor binding

• Published in: Protein science Vol. 33; no. 4; pp. e4974 - n/a
• Main Authors: Tomris, Ilhan, Woude, Roosmarijn, Paiva Froes Rocha, Rebeca, Torrents de la Peña, Alba, Ward, Andrew B., Vries, Robert P.
• Format: Journal Article
• Language: English
• Published: Hoboken, USA John Wiley & Sons, Inc 01.04.2024; Wiley Subscription Services, Inc
• Subjects: Antibodies; attachment protein; Binding; Biology; Env protein; Fluorescence; GFP; hemagglutinin; Hemagglutinins; Influenza A; influenza a virus; Molecular biology; multivalency; Polysaccharides; Polysaccharides - metabolism; Protein Binding; Proteins; Receptors; receptor‐binding; Recombinant Proteins - metabolism; Recombinants; SARS‐CoV‐2; Severe acute respiratory syndrome coronavirus 2; Spike Glycoprotein, Coronavirus - chemistry; Viral diseases; Viral envelope proteins; Viral Envelope Proteins - chemistry; Viruses; GFP; receptor‐binding; SARS‐CoV‐2; attachment protein; hemagglutinin; influenza a virus; multivalency
• ISSN: 0961-8368; 1469-896X
• 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.