Chemical characterization of cell-CAM 105, a cell-adhesion molecule isolated from rat liver membranes.
Cell-CAM 105, a glycoprotein that is involved in recognition and adhesion between isolated rat hepatocytes in vitro, was purified to homogeneity by a combination of immunoaffinity chromatography, gel-exclusion chromatography and ion-exchange chromatography. Electrophoretic, compositional and enzymic analyses were performed and the glycoprotein was shown to consist of two peptide chains, of apparent Mr 110,000 and 105,000 respectively, that are glycosylated to similar extents. Carbohydrate analyses demonstrated the presence of sialic acid, galactose, mannose, fucose and glucosamine, but no galactosamine, indicating that only N-linked oligosaccharides occurred. The total content of carbohydrate amounted to 33%. Peptide mapping indicated that the two peptide chains were structurally very similar. After incubation of cultured hepatocytes with [32P]Pi, phosphorylated cell-CAM 105 could be isolated. Both peptide chains were labelled and phospho-amino-acid analysis demonstrated that serine residues had become phosphorylated. A significant feature of cell-CAM 105 was a susceptibility to autolytic degradation that was difficult to inhibit. The major degradation products had apparent Mr 90,000 and 70,000, respectively. The effect of purified cell-CAM 105 on cell-cell adhesion of re-aggregating hepatocytes was studied. A significant inhibition was observed, indicating that the protein is directly involved in intercellular adhesion of these cells.
D000251:Adenosine Triphosphatases; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D015703:Antigens, CD; D002241:Carbohydrates / Q000032:analysis; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules; D002462:Cell Membrane / Q000032:analysis; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006026:Glycoside Hydrolases; D008099:Liver / Q000032:analysis* / Q000166:cytology; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008562:Membrane Glycoproteins / Q000302:isolation & purification; D009439:Neuraminidase; D010446:Peptide Fragments / Q000032:analysis; D010710:Phosphates / Q000032:analysis; D010766:Phosphorylation; D051381:Rats
D000596:Amino Acids; D015703:Antigens, CD; C069847:CD66 antigens; D002241:Carbohydrates; D015815:Cell Adhesion Molecules; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D010710:Phosphates; D006026:Glycoside Hydrolases; D009439:Neuraminidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D000251:Adenosine Triphosphatases