Characterization of a membrane-associated glycoprotein complex implicated in cell adhesion to fibronectin.
We have characterized a 140-kDa glycoprotein complex purified by a monoclonal antibody and implicated in cell adhesion to the extracellular molecule fibronectin. Three major polypeptide components were purified by monoclonal antibody JG22E, which had apparent molecular weights of 155,000 (band 1), 135,000 (band 2), and 120,000 (band 3). In two-dimensional gel electrophoresis, each subunit migrated as either a broad band or a series of spots at acidic isoelectric points. After treatment with neuraminidase, the spots became focused around pH 6.2 (band 1), pH 5.6 (band 2), and pH 5.3 (band 3). These three major bands were compared by two-dimensional peptide mapping in a series of pairwise combinations and were found to be distinct proteins. In sucrose gradients, these proteins co-migrated as a complex sedimenting at approximately 8.4 S either before or after affinity purification, whereas separated subunits migrated at 4.7 to 5.8 S. Amino acid analysis revealed no detectable hydroxyproline and a composition characterized by a substantial number of cysteine residues compared to the average protein. Our results suggest that a noncovalent complex of structurally distinct glycoproteins is involved in adhesive interactions of fibronectin with cells.
D000596:Amino Acids / Q000032:analysis; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000302:isolation & purification; D002448:Cell Adhesion; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D003101:Collodion; D004591:Electrophoresis, Polyacrylamide Gel; D005353:Fibronectins / Q000378:metabolism*; D006023:Glycoproteins / Q000302:isolation & purification* / Q000502:physiology; D007120:Immunochemistry; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000502:physiology
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000941:Antigens; D005353:Fibronectins; D006023:Glycoproteins; D046911:Macromolecular Substances; D008565:Membrane Proteins; D003101:Collodion