title: Fe3+(2)-transferrin and Fe3+(2)-asialotransferrin deliver iron to hepatocytes by an identical mechanism.
abstract: We have been unable to demonstrate the unequivocal presence of transferrin receptors on rat hepatocytes. The binding and presumed internalisation of 125I-Fe3+(2)-transferrin by freshly isolated hepatocytes was only partially inhibited by up to a 10(4)-fold molar excess of unlabelled ligand and was virtually insensitive to chloroquine. There would appear to be only a weak association between this ligand and some component of the hepatocyte cell surface. These results were not compatible with the commitment of Fe3+(2)-transferrin to either a receptor-mediated endocytotic pathway nor to a rapid recycling pathway through sorting endosomes. Desialylation of the biantennary oligosaccharide side chains of Fe3+(2)-transferrin engendered a low affinity (Kd greater than or equal to 0.25 microM) for the asialoglycoprotein receptor. 125I-Fe3+(2)-asialotransferrin was only superficially internalised by isolated hepatocytes but this was characteristic of ligands for the asialoglycoprotein receptor which have only biantennary (or single triantennary) side chains. This was also incompatible with the delivery of the ligand to sorting endosomes where the release of iron has been presumed to occur. Despite the different properties of the two ligands, the rates of iron uptake from 59Fe3+(2)-transferrin and 59Fe3+(2)-asialotransferrin were identical, suggesting a common mechanism for the translocation of iron across the plasma (or possibly endosomal) membrane such as a transmembrane oxidoreductase. Competition studies with unlabelled ligand or impermeable ferric ammonium citrate gave an IC50 of 1-15 micrograms Fe3+/ml for this process. The absence of transferrin receptor from the surface of the terminally differentiated, quiescent hepatocyte would be compatible with the dual roles suggested for transferrin as an iron transport protein and as a growth factor. The release of iron at the hepatocyte cell surface would effectively uncouple the two functions and render the hepatocyte unresponsive to growth stimulation by transferrin.
authors.affiliation: Department of Surgery, St George's Hospital Medical School, London, England.
authors.forename: R J
authors.initials: RJ
authors.lastname: Sharma
authors.forename: D A
authors.initials: DA
authors.lastname: Grant
mesh_terms: D000818:Animals; D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins; D001667:Binding, Competitive; D002478:Cells, Cultured; D002738:Chloroquine / Q000494:pharmacology; D007501:Iron / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D011971:Receptors, Immunologic / Q000378:metabolism; D014168:Transferrin / Q000031:analogs & derivatives* / Q000378:metabolism*
chemical_list: D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins; D008024:Ligands; D011971:Receptors, Immunologic; D014168:Transferrin; C017031:asialotransferrins; D002738:Chloroquine; D007501:Iron