Site-Specific Steric Control of SARS-CoV-2 Spike Glycosylation

Biochemistry. 2021 Jul 13;60(27):2153-2169. doi: 10.1021/acs.biochem.1c00279. Epub 2021 Jul 2.


A central tenet in the design of vaccines is the display of native-like antigens in the elicitation of protective immunity. The abundance of N-linked glycans across the SARS-CoV-2 spike protein is a potential source of heterogeneity among the many different vaccine candidates under investigation. Here, we investigate the glycosylation of recombinant SARS-CoV-2 spike proteins from five different laboratories and compare them against S protein from infectious virus, cultured in Vero cells. We find patterns that are conserved across all samples, and this can be associated with site-specific stalling of glycan maturation that acts as a highly sensitive reporter of protein structure. Molecular dynamics simulations of a fully glycosylated spike support a model of steric restrictions that shape enzymatic processing of the glycans. These results suggest that recombinant spike-based SARS-CoV-2 immunogen glycosylation reproducibly recapitulates signatures of viral glycosylation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • COVID-19 / genetics*
  • COVID-19 / immunology
  • COVID-19 / virology
  • COVID-19 Vaccines / genetics
  • COVID-19 Vaccines / immunology
  • Chlorocebus aethiops
  • Glycosylation
  • Humans
  • Molecular Dynamics Simulation
  • Protein Binding / genetics
  • Protein Conformation*
  • SARS-CoV-2 / genetics
  • SARS-CoV-2 / pathogenicity
  • SARS-CoV-2 / ultrastructure*
  • Spike Glycoprotein, Coronavirus / chemistry
  • Spike Glycoprotein, Coronavirus / genetics
  • Spike Glycoprotein, Coronavirus / immunology
  • Spike Glycoprotein, Coronavirus / ultrastructure*
  • Vero Cells


  • COVID-19 Vaccines
  • Spike Glycoprotein, Coronavirus
  • spike protein, SARS-CoV-2