Binding and endocytosis of glycoproteins and neoglycoproteins by isolated rabbit hepatocytes.
Rabbit hepatocytes were isolated by a collagenase perfusion technique, and used to study the binding and endocytosis of the glycoprotein, asialo-orosomucoid, and the neoglycoprotein, Gal39-bovine serum albumin. Both of these proteins contain exposed galactosyl residues, and were avidly bound by the lectin on the hepatic parenchymal cell surface. Steady state and kinetic experiments performed at 2 degrees C and at 37 degrees C revealed the presence of two apparent classes of binding sites totalling 4.7 X 10(5) sites/cell at 2 degrees C, and 6.3 X 10(5) sites/cell at 37 degrees C. At 37 degrees C, both classes of sites participated in internalization of bound ligand. The cells were capable of internalizing about 60 000 molecules/min per cell. The process appeared to be first-order, with a rate constant k = 0.098 min-1 and t1/2 = 7.1 +/- 0.6 min. Binding could be inhibited by galactose-containing compounds, EGTA, and by anti-(hepatic lectin) immunoglobulin G. The inhibition by antibody appeared to be reversible upon removal of antibody-containing medium.
D000818:Animals; D000906:Antibodies / Q000276:immunology; D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins; D004705:Endocytosis; D005690:Galactose; D006023:Glycoproteins / Q000378:metabolism*; D066298:In Vitro Techniques; D007700:Kinetics; D008024:Ligands; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D008297:Male; D008954:Models, Biological; D009961:Orosomucoid / Q000031:analogs & derivatives* / Q000378:metabolism; D011485:Protein Binding; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000276:immunology / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000378:metabolism*; D013696:Temperature
D000906:Antibodies; D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins; D006023:Glycoproteins; D008024:Ligands; D009961:Orosomucoid; D011956:Receptors, Cell Surface; C017030:asialoorosomucoid; C039136:bovine serum albumin-galactose (39); D012710:Serum Albumin, Bovine; D005690:Galactose