Fig. 1.

RP-HPLC of PNGase A-released pyridylaminated N-glycans from Caenorhabditis elegans embryos. Chromatograms for the wildtype N2, mutant e2144, and t3208 strains are shown annotated with dextran hydrolysate as external calibrant (in glucose units, g.u.) and with the structures found on the basis of MS, MS/MS, and digestion data; glycans are depicted according to the Standard Nomenclature for Glycans as shown. There are only minor differences in the N-glycans detected in the three strains, for example, some core α1,3-fucosylated Hex_3–5HexNAc₂Fuc_1–3 structures were not detected in the mutants. For clarity, the oligomannosidic structures are shown in the upper panel only, the phosphorylcholine-modified ones in the upper and middle panels, and the various core-modified paucimannosidic ones in the lower panel. See supplemental Fig. S2 for MS of individual fractions of N-glycans from N2 embryos and supplemental Table S1 for a full list. Previous studies have shown that different pyridylaminated Man_6–8GlcNAc₂ and Man_2–3GlcNAc₂Fuc₁ isomers have distinct RP-HPLC retention times and fragmentation patterns (33, 35, 87). PNGase, peptide:N-glycosidase; RP, reversed-phase.