Loss of the transferrin receptor during the maturation of sheep reticulocytes in vitro. An immunological approach.
Sheep reticulocyte-specific antiserum absorbed with mature sheep red cells has been used to isolate and identify reticulocyte-specific plasma-membrane proteins and to monitor their loss during incubation in vitro. Specific precipitation of labelled plasma-membrane proteins is obtained when detergent-solubilized extracts of 125I-labelled reticulocyte plasma membranes are incubated with this antiserum and Staphyloccus aureus, but not when mature-cell plasma membranes are treated similarly. During maturation of reticulocytes in vitro (up to 4 days at 37 degrees C), there is a marked decrease in the immunoprecipitable material. The anti-reticulocyte-specific antibodies have been identified as anti-(transferrin receptor) antibodies. By using these antibodies as a probe, the transferrin receptor has been shown to have a subunit molecular weight of 93 000. The data are consistent with reported molecular weights of this receptor and with the proposal that the receptor may exist as a dimer, since [125I]iodotyrosyl-peptide maps of the 93 000- and 186 000-mol.wt. components isolated are shown to be identical. Evidence is presented for the transmembrane nature of the receptor and for the presence of different binding sites for transferrin and these antibodies on the receptor.
D000818:Animals; D000954:Antigens, Surface / Q000276:immunology; D002470:Cell Survival; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D005455:Fluorescent Antibody Technique; D008565:Membrane Proteins / Q000097:blood / Q000276:immunology; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D011956:Receptors, Cell Surface / Q000276:immunology / Q000378:metabolism*; D011990:Receptors, Transferrin; D012156:Reticulocytes / Q000166:cytology* / Q000276:immunology; D012756:Sheep; D014168:Transferrin / Q000276:immunology / Q000378:metabolism*
D000954:Antigens, Surface; D008565:Membrane Proteins; D010446:Peptide Fragments; D011956:Receptors, Cell Surface; D011990:Receptors, Transferrin; D014168:Transferrin