Transferrin and iron uptake by rat reticulocytes.
The uptake of transferrin labeled with 3H and 59Fe by rat reticulocytes was studied to clarify the characteristics of the uptake process and intracellular transport. Rat reticulocytes took up transferrin in a saturable, time- and temperature-dependent manner. Scatchard analysis of the binding parameters indicated that transferrin molecules were bound to cell-surface receptors with high affinity. Monodansyl- cadaverine, a potent inhibitor of transglutaminase, reduced the amount of internalized transferrin but has no effect on the total amount of cell-associated transferrin, suggesting that transferrin is taken up by rat reticulocytes via receptor-mediated endocytosis. About 50% of the internalized 3H label was released from the cells after reincubation for 1 h in fresh medium. In contrast, no release of 59Fe label was observed. By immunoprecipitation and subsequent SDS-PAGE the released 3H-labeled product was identified as apotransferrin. Lysosomotropic reagents and a proton ionophore reduced the uptake of 59Fe. These results indicated that iron was removed from transferrin at an intracellular site in an acidic environment. The released iron was found not to associate with any intermediate ligands before it was utilized for heme synthesis in mitochondria.
Kohno
H
H
Tokunaga
R
R
D000217:Acyltransferases / Q000037:antagonists & inhibitors; D000818:Animals; D001059:Apoproteins; D001692:Biological Transport; D002103:Cadaverine / Q000031:analogs & derivatives / Q000494:pharmacology; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D004705:Endocytosis; D005089:Exocytosis; D006454:Hemoglobins / Q000378:metabolism; D007501:Iron / Q000378:metabolism*; D007700:Kinetics; D008247:Lysosomes / Q000187:drug effects; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D011956:Receptors, Cell Surface / Q000378:metabolism; D011990:Receptors, Transferrin; D012156:Reticulocytes / Q000378:metabolism*; D014168:Transferrin / Q000378:metabolism*; D011503:Transglutaminases
D001059:Apoproteins; D006454:Hemoglobins; D011956:Receptors, Cell Surface; D011990:Receptors, Transferrin; D014168:Transferrin; C028284:apotransferrin; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D007501:Iron; D000217:Acyltransferases; D011503:Transglutaminases; C008542:monodansylcadaverine; D002103:Cadaverine