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. 2022 Feb 24;2(3):631–645. doi: 10.1021/jacsau.1c00529

Figure 4.

Figure 4

Glycosylation of MUC14TR-T3*S14*T15* by GalNAc-T4 and the GalNAc-T4D459H mutant. Table S5 details the experimental conditions. (A) Scheme of the MUC14TR-T3*S14*T15* glycosylation event catalyzed by GalNAc-T4. The glycosylation sites are displayed in red. (B,D) 1H/15N-HSQC of the MUC14TR-T3*S14*T15* product in the presence of GalNAc-T4 (B) and GalNAc-T4D459H (D). In black, only with MnCl2, and in magenta and blue, after addition of UDP-GalNAc (in excess) at 2 and 24 h, respectively. The arrows indicate CSP that occurs as result of GalNAc additions at S4 (gray arrows) and at T8 (yellow arrows). The * labeling in the amino acid on the spectra indicates glycosylation. (C,E) Time course of relative % of glycosylation at S4 and T8 residues by GalNAc-T4 (C) and GalNAc-T4D459H (E) (see the Experimental Section for details).