Functional erythropoietin receptor of the cells with neural characteristics: Comparison with receptor properties of erythroid cells

Seiji Masuda, Masaya Nagao, Kyoya Takahata, Yoshihiro Konishi, F. Gallyas, Takeshi Tabira, Ryuzo Sasaki

Research output: Contribution to journalArticle

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Abstract

Radioiodinated erythropoietin (Epo) was bound specifically to the cells of two non-erythroid clonal lines, PC12 and SN6, which expressed neuronal characteristics. The binding was time-, cell number-, and dose-dependent and was reversible. Although the cloned Epo receptor from PC12 cells (derived from rat adrenal medulla) was identical to that from rat erythroid cells, significant differences in the ligand binding properties between two cell lineages were found; 1) PC12 cells had a single class of binding sites with very low affinity (Kd = 16 nM), whereas erythroid cells had two classes of binding sites with different affinities (Kd = 95 pM for high affinity sites and 1.9 nM for low affinity sites), and 2) cross-linking experiments revealed one cross-linked product of 105 kDa for PC12 cells and two products of 140 and 120 kDa for erythroid cells. Taken together with additional results, the presence of a putative accessory protein(s) that may alter the ligand binding affinity through interaction with Epo receptor is discussed. The binding of Epo to PC12 cells caused a rapid increase in the cytosolic concentration of free calcium. The presence of EGTA had no effect on the Epo binding but completely inhibited the calcium increase, indicating that Epo stimulated the calcium influx from outside of the cells. The addition of Epo to the culture media of PC12 cells elevated the intracellular concentrations of monoamines.

Original languageEnglish
Pages (from-to)11208-11216
Number of pages9
JournalJournal of Biological Chemistry
Volume268
Issue number15
Publication statusPublished - May 25 1993

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Erythropoietin Receptors
Erythroid Cells
PC12 Cells
Erythropoietin
Calcium
Rats
Binding Sites
Ligands
Adrenal Medulla
Egtazic Acid
Accessories
Cell Lineage
Culture Media
Cell Count
Proteins
Experiments

ASJC Scopus subject areas

  • Biochemistry

Cite this

Functional erythropoietin receptor of the cells with neural characteristics : Comparison with receptor properties of erythroid cells. / Masuda, Seiji; Nagao, Masaya; Takahata, Kyoya; Konishi, Yoshihiro; Gallyas, F.; Tabira, Takeshi; Sasaki, Ryuzo.

In: Journal of Biological Chemistry, Vol. 268, No. 15, 25.05.1993, p. 11208-11216.

Research output: Contribution to journalArticle

Masuda, Seiji ; Nagao, Masaya ; Takahata, Kyoya ; Konishi, Yoshihiro ; Gallyas, F. ; Tabira, Takeshi ; Sasaki, Ryuzo. / Functional erythropoietin receptor of the cells with neural characteristics : Comparison with receptor properties of erythroid cells. In: Journal of Biological Chemistry. 1993 ; Vol. 268, No. 15. pp. 11208-11216.
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