Figure 5: CD209L is N-glycosylated and N-glycosylation modulates its interaction with SARS-CoV2 Spike Protein.

(A) HCD MS/MS spectrum of m/z 1505.1470, assigned as the [M + 2H]²⁺ of the glycopeptide⁸⁷DAIYQNLTQLK⁹⁷ + HexNAc₂Hex₈, corresponding to CD209L site N92 bearing a Man₈ glycan. Stepped-collision energy (15/25/35% NCE) was used. The fragment ions do not retain the glycan moiety. p = peptide, N = N-acetylhexosamine (HexNAc), H = hexose (Hex). (B) Dot blot of CD209L/HEK-293 cell lysate under control conditions (no treatment), or treated with PNGase F or Endo H. Dot blot of HEK-293 cell lysate was used as a negative control. Following treatment of lysate, the spike protein receptor-binding domain (S-RBD-HIS) was incubated with the immobilized lysate, followed by detection with anti-HIS antibody. (C) Quantification of blots is shown. AU, arbitrary unit. Image J software was used to quantify the dot blots