Nanoluciferase complementation-based biosensor reveals the importance of N- linked glycosylation of SARS-CoV-2 Spike for viral entry [post]

Taha Azad, Ragunath Singaravelu, Zaid Taha, Stephen Boulton, Mathieu J.F. Crupi, Nikolas T. Martin, Taylor Jamieson, Joanna Poutou, Mina Ghahremani, Adrian Pelin, Kazem Nouri, Rozanne Arulanandam (+7 others)
2020 unpublished
The ongoing COVID-19 pandemic has highlighted the' immediate need for the development of antiviral therapeutics targeting different stages of the SARS-CoV-2 lifecycle. We developed a bioluminescence-based biosensor to interrogate the interaction between the SARS-CoV-2 viral spike protein and its host entry receptor, angiotensin-converting enzyme 2 (ACE2). The biosensor assay is based on a Nanoluciferase complementation reporter, composed of two subunits, Large BiT and Small BiT, fused to the
more » ... iT, fused to the spike receptor-binding domain (RBD) of the SARS-CoV-2 S protein and ACE2 ectodomain, respectively. Using this biosensor, we uncovered a critical role for glycosylation of asparagine residues within the RBD in mediating successful binding to the cellular ACE2 receptor and subsequent virus infection. Our findings support RBD glycosylation as a therapeutic and vaccine target to blunt SARSCoV- 2 infections.
doi:10.21203/rs.3.rs-58455/v1 fatcat:gq7dus234ra4djn3wftqdak7lu