Structural analysis of O-linked sugar chains in human blood clotting factor IX.
Type and structural analysis of O-linked sugar chains in human blood clotting factor IX was performed by the pyridylamination method developed for O-linked sugar chains [Kuraya, N. & Hase, S. (1992) J. Biochem. 112, 122-126]. O- and N-linked sugar chains were released with hydrazine, and then N-acetylated, followed by pyridylamination. The type of sugar chain was determined by reducing-end analysis of the pyridylaminated (PA-) sugar chains. Sugar chains with PA-GalNAc at the reducing terminal and that with PA-Fuc [Nishimura, H. et al. (1992) J. Biol. Chem. 267, 17520-17525] were obtained besides known sugar chains with PA-Glc from the Xyl-Glc-Ser type and those with PA-GlcNAc from asparagine-linked sugar chains. The sugar chains with PA-GalNAc were identified as mono- and disialyl Gal beta 1-3GalNAc by two-dimensional HPLC mapping. The structure of the sugar chain with PA-Fuc was Neu5Ac alpha 2-6Gal beta 1-4GlcNAc beta 1-3Fuc, as determined by exoglycosidase digestion, methylation analysis, and Smith degradation.
Kuraya
N
N
Department of Chemistry, Osaka University College of Science.
Omichi
K
K
Nishimura
H
H
Iwanaga
S
S
Hase
S
S
D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D005591:Chemical Fractionation; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D005164:Factor IX / Q000737:chemistry*; D006801:Humans; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000097:blood / Q000737:chemistry*
D009844:Oligosaccharides; D005164:Factor IX