Figure 3
From: Fluorescent glycan fingerprinting of SARS2 spike proteins
Fingerprinting N-glycans released from various SARS2 Spike proteins with ST6Gal1 (red) and FUT8 (green). N-glycans of various recombinant spike proteins released by PNGase F were labeled by ST6Gal1 and FUT8 with (+) or without (−) pretreatment of a neuraminidase (Neu). Labeled samples were separated on a 17% SDS-PAGE gel and imaged with traditional protein imaging (upper panels) or fluorescent imaging (middle and lower panel in different contrasts). Bands in red are complex or hybrid glycans labeled by ST6Gal1 (via the incorporation of Cy5-Neu5Ac). Bands in green are oligo-mannose glycans labeled by FUT8 (via the incorporation of Alexa Fluor 555-conjugated fucose). For FUT8 labeling, MGAT1 and UDP-GlcNAc were also included in the labeling mix to convert the oligo-mannose glycans to the substrates for FUT8. Labeled glycans from ribonuclease B, A2G1S1, A2G2S(6)1 and FA2G2S(6)1 were run as references in (A) and a glycan ladder with 10 labeled standard glycans was run in (B). RS, RBD domain of SARS2 Spike protein expressed in Sf21 cells; RC, RBD domain of SARS2 Spike protein expressed in CHO cells; RH, RBD domain of SARS2 Spike protein expressed in HEK293 cells; SC, whole SARS2 Spike protein expressed in CHO cells; S1H, SARS2 S1 protein expressed in HEK293 cells; SH, SARS2 Spike protein expressed in HEK293 cells.
