Deducing the N- and O-glycosylation profile of the spike protein of novel coronavirus SARS-CoV-2

Asif Shajahan; Nitin T. Supekar; Anne S. Gleinich; Parastoo Azadi

The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.01.020966 doi: bioRxiv preprint Figure 1 : The SARS-Cov-2 spike proteins recombinatly expressed on HEK293 supernatant were fractionated through SDA-PAGE, subsequently digested by proteases anc analyzed by nLC-NSI-MS/MS. The expression of SARS-Cov-2 spike protein subunits S1 and S2 on HEK 293 culture supernatant showed higher molecular weight upon SDS-PAGE than expected, because of glycosylation. Thus, the gel bands corresponding to the molecular weight of 200 kDa to 100 kDa for S1 and 150 kDa to 80 kDa for S2 were cut, proteins were lysed after reduction-alkylation and analyzed by LC-MS/MS (created with biorender.com). The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.01.020966 doi: bioRxiv preprint Figure 2 : Glycosylation profile on coronavirus SARS-CoV-2 characterized by high-resolution LC-MS/MS. About 17 N-glycosylation sites were found occupied out of 22 potential sites along with two O-glycosylation sites bearing core-1 type O-glycans. Some N-glycosylation sites were partially glycosylated. author/funder. All rights reserved. No reuse allowed without permission.