HDX-MS Protocol

HDX MS – sample preparation for the analysis of SARS-CoV-2 S-protein binding to recombinant ACE-2 entry receptor ectodomain (and mAb when available)

Previous cryo EM studies have shown that the SARS-CoV-2 S protein binds with high affinity to the ACE-2 receptor (15nM affinity)1. The structure of the S protein RBD (receptor binding domain) bound to ACE2 has also been determined2. HDX-MS experiments can be carried out with SARS-CoV-2 S (Receptor binding domain), ACE2 and mAbs (when these become available), and this would be complementary to the work proposed by Michal Sharon’s group. HDX could also be used to investigate the effects of glycosylation on binding3.

1. Wrapp, D., Wang, N., Kizzmekia, S. et al. Cryo-EM structure of the 2019-nCoV spike in the prefusion conformation Science (2020). https//doi.org/10.1126/science.abb250
2. Lan, J., Ge, J., Yu, J. et al. Structure of the SARS-CoV-2 spike receptor-binding domain bound to the ACE2 receptor. Nature (2020). https://doi.org/10.1038/s41586-020-2180-5
3. Zhang Y., Zhao W., Mao S. et al. Site-specific N-glycosylation Characterization of Recombinant SARS-CoV-2 Spike Proteins using High-Resolution Mass Spectrometry. bioRxiv preprint (2020) https://doi.org/10.1101/2020.03.28.013276