Galactose-specific recognition system of mammalian liver: receptor distribution on the hepatocyte cell surface.
An isolated perfused liver system was used to study the distribution of asialoglycoprotein (ASGP) binding sites on rat hepatocyte cell surfaces. The number of surface receptors was quantitated by monitoring clearance of 125I-labeled ligands from the perfusate medium under two conditions that blocked their internalization: low temperature (less than 5 degrees C) or brief formaldehyde fixation. The cell surface distribution of binding sites was visualized in the electron microscope with either asialoorosomucoid covalently coupled to horseradish peroxidase (ASOR-HRP) or lactosaminated ferritin (Lac-Fer), both of which were bound with similar kinetics and to similar extents as ASOR itself. At low temperature or after prefixation, ASGP binding sites were present over much of the sinusoidal cell surface, but were concentrated most heavily over coated pits. Quantitation of ligand distribution at 4 degrees C with Lac-Fer gave an approximately 70-fold greater density of ferritin particles over coated membrane than over uncoated regions. We obtained no evidence for gradual movement of ASGP receptors into or out of coated pits within the time-course of our experiments. Finally, the number and distribution of cell surface binding sites was unaffected by previous exposure to ASOR or by inhibition of endocytic vesicle-lysosome fusion and ASOR degradation at 16 degrees C.
D000606:Amino Sugars / Q000378:metabolism; D000818:Animals; D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins; D002462:Cell Membrane / Q000032:analysis / Q000648:ultrastructure; D004705:Endocytosis; D005293:Ferritins / Q000378:metabolism; D006735:Horseradish Peroxidase; D008099:Liver / Q000032:analysis* / Q000648:ultrastructure; D009961:Orosomucoid / Q000031:analogs & derivatives / Q000378:metabolism; D051381:Rats; D011956:Receptors, Cell Surface / Q000032:analysis*; D013696:Temperature
D000606:Amino Sugars; D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins; D009961:Orosomucoid; D011956:Receptors, Cell Surface; C017030:asialoorosomucoid; C032233:lactosamine; D005293:Ferritins; D006735:Horseradish Peroxidase