Occurrence and structural analysis of highly sulfated multiantennary N-linked glycan chains derived from a fertilization-associated carbohydrate-rich glycoprotein in unfertilized eggs of Tribolodon hakonensis.
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-beta-galactosidase and peptide:N-glycosidase F) digestions, and instrumental analyses (1H-NMR spectroscopy and FAB-MS) which revealed the novel repeating sulfated carbohydrate sequences, +/- Gal beta 1-->4Gal beta 1[-->(HSO3-->6)GlcNAc beta 1-->3(+/- Gal beta 1-->4)Gal beta 1]n--> (see Structure I; p + q + r + s approximately 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 -->4GlcNAc beta 1-->3(Gal beta 1-->4)Gal beta 1-->. [formula: see text]
Taguchi
T
T
Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Japan.
Iwasaki
M
M
Muto
Y
Y
Kitajima
K
K
Inoue
S
S
Khoo
K H
KH
Morris
H R
HR
Dell
A
A
Inoue
Y
Y
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002850:Chromatography, Gel; D002855:Chromatography, Thin Layer; D003530:Cyprinidae; D005260:Female; D029941:Fish Proteins; D006023:Glycoproteins / Q000737:chemistry*; D006834:Hydrazines / Q000494:pharmacology; D009682:Magnetic Resonance Spectroscopy; D008745:Methylation; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000032:analysis; D009608:Nitrous Acid / Q000494:pharmacology; D009844:Oligosaccharides / Q000032:analysis / Q000737:chemistry; D010063:Ovum / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry*; D017421:Sequence Analysis; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013431:Sulfates / Q000737:chemistry
D029941:Fish Proteins; D006023:Glycoproteins; D006834:Hydrazines; D009005:Monosaccharides; D009844:Oligosaccharides; D011134:Polysaccharides; D013431:Sulfates; C061150:hyosophorin protein, Fish; C029424:hydrazine; D009608:Nitrous Acid