Determining the N-terminus of RBD Protein
by EThcD. (A) Predicted
N-terminal peptide as found in M68, M69, and original Ragon Institute
constructs. (B) N-terminal peptide found in M67 and original Mt. Sinai
constructs containing an additional Gln residue. This Gln residue
was modified to a pyroGlu residue, as noted by the yellow box. (C)
N-terminal peptide found in M68, M69, and original Ragon Institute
constructs containing an additional Ser residue. (D) N-terminal peptide
found in M68, M69, and original Ragon Institute constructs containing
additional Ser-Ala-Ser residues. Carbamidomethylated Cys is noted
in blue, and deamidated Asn residues are noted by gray boxes. Observed c/z ions are noted by red flags between
residues, while observed b/y ions are noted by blue
flags. All peptides were found to be O-glycosylated with Hex(1)HexNAc(1)NeuAc(2)
on the first Thr residue (T5 of RBD; T323 of full-length spike) as
noted by the yellow box with glycan pictogram. The complementary ion
pairs resulting from the backbone cleavage of T5/E6 and E6/S7 confirm
the glycosylation localization.