This study represents the first detailed investigation of the nature of highly sulfated (keratan-sulfate-like) complex-type asparagine-linked glycans having a tetraantennary core structure and shows the effectiveness of fast-atom-bombardment mass spectrometric (FAB-MS) methods incorporating derivatization and mild methanolysis for analyzing such complex types of sulfated glycans. The structure of the N-glycan chains was unambiguously established by a combination of compositional analysis, methylation analysis, mild methanolysis for desulfation, hydrazinolysis/nitrous acid deamination, enzymatic (endo-β-galactosidase and peptide:N-glycosidase F) digestions, and instrumental analyses (1H-NMR spectroscopy and FAB-MS) which revealed the novel repeating sulfated carbohydrate sequences, ±Galβ1→4Galβ1[→4(HSO 3→6)GlcNAcβ1→3(±Galβ1-4)Galβ1] n→ (see Structure 1: p+q+r+s≈14). This sequence is unique in: (a) the skeletal structure is similar to that of keratan sulfate but is completely devoid of 6-O-sulfated Gal residues and (b) the presence of branched Gal residues in the sequence →4GlcNAcβ1→3(Galβ1→4)Galβ1→. (Formula Presented).
- Fast-atom-bombardment mass spectrometry
- Highly sulfated N-linked glycan
- Keratan-sulfate-like N-linked glycan
- Tetraantennary glycan
ASJC Scopus subject areas