Identification and analysis of human erythropoietin receptors on a factor-dependent cell line, TF-1
We have recently established a novel cell line, TF-1, from bone marrow cells of a patient with erythroleukemia, that showed an absolute growth dependency on each of three hematopoietic growth factors: erythropoietin (EPO) granulocyte-macrophage colony-stimulating factor (GM-CSF), and interleukin 3 (IL-3). EPO stimulated the proliferation of TF-1 cells even at the physiologic concentration (0.03 U/mL). We performed binding experiments on TF-1 cells using radioiodinated EPO. The binding of radioiodinated EPO to TF-1 was specific, time- and temperature-dependent, and saturable. Scatchard analysis of the saturation binding data suggested the existence of a single class of binding sites (kd = 0.40 nmol/L; number of binding sites = 1,630 per cell). TF-1 cells were usually maintained in RPMI 1640 containing 10% fetal bovine serum and 5 ng/mL GM-CSF. The kd and the number of the EPO receptors were not changed by incubating the cells with IL-3, although culturing the cells in the presence of EPO resulted in down-modulation of EPO receptors. The chemical cross-linking study demonstrated that two molecules with apparent molecular weights of 105 kilodalton (Kd) and 90 Kd were the binding components of EPO. Present data suggest that human EPO receptors are very similar to the previously reported murine EPO receptors.
- Research Organization:
- Univ. of Tokyo (Japan)
- OSTI ID:
- 6327101
- Journal Information:
- Blood; (United States), Vol. 73:2
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ERYTHROPOIETIN
RECEPTORS
GROWTH FACTORS
BIOCHEMICAL REACTION KINETICS
TUMOR CELLS
CELL PROLIFERATION
CROSS-LINKING
IODINE ISOTOPES
LYMPHOKINES
MAN
MOLECULAR WEIGHT
TEMPERATURE DEPENDENCE
TIME DEPENDENCE
TRACER TECHNIQUES
ANIMAL CELLS
ANIMALS
CHEMICAL REACTIONS
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
MAMMALS
MEMBRANE PROTEINS
MITOGENS
ORGANIC COMPOUNDS
POLYMERIZATION
PRIMATES
PROTEINS
REACTION KINETICS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques