Two Receptor Binding Strategy of SARS-CoV-2 Is Mediated by Both the N-Terminal and Receptor-Binding Spike Domain

Michele Monti*, Edoardo Milanetti, Myrthe T Frans, Mattia Miotto, Lorenzo Di Rienzo, Maksim V Baranov, Giorgio Gosti, Arun Kumar Somavarapu, Madhu Nagaraj, Thaddeus W Golbek, Emiel Rossing, Sam J Moons, Thomas J Boltje, Geert van den Bogaart, Tobias Weidner, Daniel E Otzen, Gian Gaetano Tartaglia, Giancarlo Ruocco, Steven J Roeters*

*Corresponding author voor dit werk

OnderzoeksoutputAcademicpeer review

3 Citaten (Scopus)
47 Downloads (Pure)


It is not well understood why severe acute respiratory syndrome (SARS)-CoV-2 spreads much faster than other β-coronaviruses such as SARS-CoV and Middle East respiratory syndrome (MERS)-CoV. In a previous publication, we predicted the binding of the N-terminal domain (NTD) of SARS-CoV-2 spike to sialic acids (SAs). Here, we experimentally validate this interaction and present simulations that reveal a second possible interaction between SAs and the spike protein via a binding site located in the receptor-binding domain (RBD). The predictions from molecular-dynamics simulations and the previously-published 2D-Zernike binding-site recognition approach were validated through flow-induced dispersion analysis (FIDA)─which reveals the capability of the SARS-CoV-2 spike to bind to SA-containing (glyco)lipid vesicles, and flow-cytometry measurements─which show that spike binding is strongly decreased upon inhibition of SA expression on the membranes of angiotensin converting enzyme-2 (ACE2)-expressing HEK cells. Our analyses reveal that the SA binding of the NTD and RBD strongly enhances the infection-inducing ACE2 binding. Altogether, our work provides in silico, in vitro, and cellular evidence that the SARS-CoV-2 virus utilizes a two-receptor (SA and ACE2) strategy. This allows the SARS-CoV-2 spike to use SA moieties on the cell membrane as a binding anchor, which increases the residence time of the virus on the cell surface and aids in the binding of the main receptor, ACE2, via 2D diffusion.

Originele taal-2English
Pagina's (van-tot)451-464
Aantal pagina's14
TijdschriftThe Journal of Physical Chemistry. B: Materials, Surfaces, Interfaces, & Biophysical
Nummer van het tijdschrift2
Vroegere onlinedatum8-jan.-2024
StatusPublished - feb.-2024


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