Identification and quantification of the rat hepatocyte asialoglycoprotein receptor.
The asialoglycoprotein receptor from rat liver was purified by solubilization and affinity chromatography on asialoorosomucoid-Sepharose. The preparation yielded four distinct polypeptides of Mr 40,000-120,000. We prepared a monoclonal antibody that both immunoprecipitates solubilized receptor activity and blocks the binding of galactose-terminal glycoproteins to immobilized receptor. The monoclonal antibody and a rabbit antireceptor antiserum immunoprecipitated all four polypeptide species. Peptide analysis by two-dimensional chromatography of the individual 125I-labeled species showed nearly identical patterns, which also suggested that the four polypeptides have a similar primary structure. To identify and quantitate the asialoglycoprotein receptor on the hepatocyte cell surface, intact cells were iodinated with lactoperoxidase, and the solubilized membranes were treated with antireceptor antibody. The Mr 55,000 and Mr 65,000 species were the major species found. Our results suggest that the Mr of the surface receptor is at least 55,000 and that it comprises between 1-2% of the iodinated hepatocyte surface protein.
D000818:Animals; D037263:Asialoglycoprotein Receptor; D002999:Clone Cells / Q000276:immunology; D006023:Glycoproteins; D006822:Hybrid Cells / Q000276:immunology; D007158:Immunologic Techniques; D008099:Liver / Q000032:analysis*; D046911:Macromolecular Substances; D008970:Molecular Weight; D009961:Orosomucoid; D010446:Peptide Fragments / Q000032:analysis; D051381:Rats; D011956:Receptors, Cell Surface / Q000032:analysis* / Q000276:immunology
D037263:Asialoglycoprotein Receptor; D006023:Glycoproteins; D046911:Macromolecular Substances; D009961:Orosomucoid; D010446:Peptide Fragments; D011956:Receptors, Cell Surface