Transferrin uptake and release by reticulocytes treated with proteolytic enzymes and neuraminidase.
The mechanism of transferrin uptake by reticulocytes was investigated using rabbit transferrin labelled with 125I and 59Fe and rabbit reticulocytes which had been treated with trypsin, Pronase or neuraminidase. Low concentrations of the proteolytic enzymes produced a small increase in transferrin and iron uptake by the cells. However, higher concentrations or incubation of the cells with the enzymes for longer periods caused a marked fall in transferrin and iron uptake. This fall was associated with a reduction in the proportion of cellular transferrin which was bound to a cell membrane component solubilized with the non-ionic detergent, Teric 12A9. The effect of trypsin and Pronase on transferrin release from the cells was investigated in the absence and in the presence of N-ethylmaleimide which inhibits the normal process of transferrin release. It was found that only a small proportion of transferrin which had been taken up by reticulocytes at 37 degrees C but nearly all that taken up 4 degrees C was released when the cells were subsequently incubated with trypsin plus N-ethylmaleimide, despite the fact that about 80% of the 59Fe in the cells was released in both instances. Neuraminidase produced no change in transferrin and iron uptake by the cells. These experiments provide evidence that transferrin uptake by reticulocytes requires interaction with a receptor which is protein in nature and that following uptake at 37 degrees C, most of the transferrin is located at a site unavailable to the action of proteolytic enzymes. The results support the hypothesis that transferrin enters reticulocytes by endocytosis.
D000818:Animals; D001692:Biological Transport; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D003902:Detergents / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D007467:Iodoproteins / Q000097:blood; D007501:Iron / Q000097:blood; D007700:Kinetics; D008854:Microscopy, Electron; D009439:Neuraminidase / Q000494:pharmacology; D011402:Pronase / Q000494:pharmacology; D011817:Rabbits; D012156:Reticulocytes / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D014168:Transferrin / Q000378:metabolism*; D014357:Trypsin / Q000494:pharmacology
D003902:Detergents; D007467:Iodoproteins; D014168:Transferrin; D007501:Iron; D009439:Neuraminidase; D014357:Trypsin; D011402:Pronase; D005033:Ethylmaleimide