Structural study of the sugar chains of human leukocyte cell adhesion molecules CD11/CD18.
Leu-CAMs (CD11/CD18) consisting of LFA-1, Mac-1, and p150/95 are leukocyte cell surface glycoproteins that are involved in various leukocyte functions. The asparagine-linked sugar chains were released as oligosaccharides from Leu-CAMs by hydrazinolysis. About 12 mol of sugar chains was released from 1 mol of Leu-CAMs. These sugar chains were converted to radioactive oligosaccharides by reduction with sodium borotritide and separated into neutral and acidic fractions by paper electrophoresis. All of the acidic oligosaccharides were converted to neutral ones by digestion with sialidase, indicating that they are sialyl derivatives. The neutral and sialdase-treated acidic oligosaccharides were fractionated by chromatography on lectin columns followed by Bio-Gel P-4 column chromatography. Structural studies of each oligosaccharide by sequential exo- and endoglycosidase digestion and by methylation analysis revealed that Leu-CAMs contain mainly high mannose type and high molecular weight complex type sugar chains. The latter sugar chains were of bi-, tri-, and tetraantennary complex types with the Gal beta 1----4(Fuc alpha 1----3)GlcNAc beta 1----and/or the Gal beta 1----3GlcNAc beta 1----groups together with the Gal beta 1----4GlcNAc group in their outer-chain moieties. In addition to these sugar chains, a small amount of monoantennary complex type and hybrid type sugar chains was found in Leu-CAMs. Furthermore, analysis of the asparagine-linked sugar chains released from the beta-subunit of Leu-CAMs by a series of lectin chromatography showed that subunit-specific glycosylation is not observed between the alpha- and beta-subunits of Leu-CAMs.
Asada
M
M
Department of Biochemistry, University of Tokyo, Japan.
Furukawa
K
K
Kantor
C
C
Gahmberg
C G
CG
Kobata
A
A
D015703:Antigens, CD / Q000737:chemistry*; D000943:Antigens, Differentiation / Q000737:chemistry*; D001216:Asparagine; D018845:CD11 Antigens; D018821:CD18 Antigens; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D004590:Electrophoresis, Paper; D006026:Glycoside Hydrolases; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D007962:Leukocytes / Q000276:immunology; D008745:Methylation; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000302:isolation & purification*; D016028:Receptors, Leukocyte-Adhesion / Q000737:chemistry*
D015703:Antigens, CD; D000943:Antigens, Differentiation; D018845:CD11 Antigens; D018821:CD18 Antigens; D009844:Oligosaccharides; D016028:Receptors, Leukocyte-Adhesion; D001216:Asparagine; D006026:Glycoside Hydrolases