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Identification of the receptor for erythropoietin by cross-linking to Friend virus-infected erythroid cells

Journal Article · · Proc. Natl. Acad. Sci. U.S.A.; (United States)
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Erythropoietin (Epo) is a glycoprotein hormone that regulates erythroid development and interacts with surface receptors on developing erythroid cells. In this laboratory, a cell system with a relatively pure population of erythroid cells that respond to Epo has been developed. Immature erythroid cells are obtained from the spleens of mice infected with the anemia strain of Friend virus. The binding of /sup 125/I-labeled Epo (/sup 125/-Epo) to plasma membranes from these cells was studied in this investigation. /sup 125/I-Epo binding reached equilibrium within 20 min at 37/sup 0/C. Twenty percent of the receptors bound /sup 125/I-Epo with a K/sub d/ of 0.08 x 10/sup -9/ M, while the remaining receptors bound the hormone with a k/sub d/ of 0.6 x 10/sup -9/ M. In this study, a receptor for Epo was identified by cross-linking /sup 125/I-Epo to the receptor in intact cells and plasma membrane preparations using disuccinimidyl suberate. Polyacrylamide gel electrophoresis revealed two labeled bands of 100 and 85 kDa. The 85-kDa band was more heavily labeled (65%) than the 100-kDa band. Both bands were equally decreased when increasing amounts of unlabeled Epo were included in the binding mixture, indicating a specific interaction of /sup 175/I-Epo with the receptor.
Research Organization:
Vanderbilt Univ. School of Medicine, Nashville, TN
OSTI ID:
5437293
Journal Information:
Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 84:12; ISSN PNASA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
BETA DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BLOOD FORMATION
BODY
CARBOHYDRATES
CELL CONSTITUENTS
CELL DIFFERENTIATION
CELL MEMBRANES
CHEMICAL REACTIONS
CROSS-LINKING
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
ERYTHROPOIESIS
ERYTHROPOIETIN
GLUCOPROTEINS
HORMONES
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPES
KINETICS
LABELLED COMPOUNDS
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
MICE
MICROORGANISMS
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
PARASITES
PEPTIDE HORMONES
POLYMERIZATION
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RECEPTORS
RODENTS
SACCHARIDES
SPLEEN
VERTEBRATES
VIRUSES