Specific binding of erythropoietin to its receptor on responsive mouse erythroleukemia cells.
Erythropoietin (Epo) is a glycoprotein factor that specifically regulates the proliferation and differentiation of erythroid progenitor cells. Here we describe the isolation of Epo-responsive mouse erythroleukemia cell line SKT6, the characterization of the specific binding of biologically active 125I-labeled human Epo (125I-Epo) to its membrane receptor, and, finally, report information concerning the molecular structure of the receptor. About 75% of erythroid colony-forming precursor cell-like colonies derived from SKT6 cells were hemoglobin-positive after 3- to 4-day exposure to Epo in methylcellulose culture. Radioiodinated Epo bound specifically to SKT6 cells, and Scatchard analysis of the data showed a high affinity for 125I-Epo (Kd = 0.15 nM) but displayed only a small number of specific receptors (approximately equal to 470 per cell). Membrane components that specifically interact with 125I-Epo were identified by covalent crosslinking with disuccinimidyl suberate, and three receptor species with apparent Mr 63,000, 94,000, and 119,000 were found in membrane from SKT6 cells, suggesting the complex structure of the receptor molecules. Specific bindings were also detected in all of the Epo-unresponsive Friend erythroleukemia cells examined, and cross-linking study revealed the presence of only the 63,000 species as a binding site.
D000818:Animals; D002455:Cell Division; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004921:Erythropoietin / Q000378:metabolism*; D007700:Kinetics; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D011956:Receptors, Cell Surface / Q000378:metabolism*; D017467:Receptors, Erythropoietin
D011956:Receptors, Cell Surface; D017467:Receptors, Erythropoietin; D004921:Erythropoietin