Structural study of N-linked oligosaccharides of human intercellular adhesion molecule-3 (CD50).
The N-linked oligosaccharides were released from purified human intercellular adhesion molecule (ICAM)-3 by hydrazinolysis. Approximately 6 mol of oligosaccharides were released from 1 mol of ICAM-3. The oligosaccharides reduced with NaB[3H]4 were separated into neutral and acidic fractions by paper electrophoresis. Most of the acidic oligosaccharides were converted to neutral ones by digestion with sialidase, indicating that they are sialyl derivatives. The neutral and sialidase-treated acidic oligosaccharides were fractionated by serial lectin column chromatography followed by Bio-Gel P-4 column chromatography. Structural studies of each oligosaccharide by sequential exo- and endo-glycosidase digestion and by methylation analysis revealed that N-linked oligosaccharides of ICAM-3 are mainly of tri- and tetra-antennary complex-type, about 60% of which contain two to three poly N-acetyllactosamine chains terminated with the type 1 structure and those without the type 1 structure per oligosaccharide. In addition, a small amount of the high mannose-type oligosaccharide with six alpha-mannose residues, which could act as a ligand for the dendritic cell-specific ICAM-3 grabbing nonintegrin, was detected.
Funatsu
O
O
Department of Biosignal Research, Tokyo Metropolitan Institute of Gerontology, Itabashi-ku, Japan.
Sato
T
T
Kotovuori
P
P
Gahmberg
C G
CG
Ikekita
M
M
Furukawa
K
K
D015703:Antigens, CD; D000943:Antigens, Differentiation; D002240:Carbohydrate Sequence; D015815:Cell Adhesion Molecules / Q000737:chemistry*; D002853:Chromatography, Liquid; D003208:Concanavalin A / Q000737:chemistry; D004590:Electrophoresis, Paper; D006026:Glycoside Hydrolases / Q000737:chemistry; D006801:Humans; D037102:Lectins / Q000737:chemistry; D007962:Leukocytes / Q000737:chemistry; D008745:Methylation; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification
D015703:Antigens, CD; D000943:Antigens, Differentiation; D015815:Cell Adhesion Molecules; C483616:ICAM3 protein, human; D037102:Lectins; D009844:Oligosaccharides; D003208:Concanavalin A; D006026:Glycoside Hydrolases