Extraction of erythropoietin from normal kidneys. [Rats, dogs]

Journal Article · Fri Sep 01 04:00:00 EDT 1978 · Endocrinology; (United States)

OSTI ID:6624019

Sherwood, J B; Goldwasser, E

Significant amounts of active erythropoietin were extracted from the kidneys of normal rats, cattle, dogs, and rabbits by homogenization of the organs in 0.1 M phosphate buffer. The mean erythropoietin activities of the extracts, as determined by the starved-rat assay, were 0.26 U/g beef kidney, 0.41 U/g dog kidney, and 0.11 U/g rat kidney. The dog kidney extracts had a mean activity of 0.35 U/g, as measured by stimulation of hemoglobin synthesis in cultured bone marrow cells (in vitro assay) and produced a dose-dependent stimulation of /sup 59/Fe incorporation into circulating red cells when assayed in polycythemic mice. Extracts of rabbit kidney cortices had a mean activity of 2.12 U/g, as measured by stimulation of hemoglobin synthesis in cultured bone marrow cells. When the dog kidney homogenate was fractionated on DEAE-cellulose, all of the erythropoietin activity was adsorbed to the exchanger in the presence of 0.01 M acetate buffer, pH 4.5, and was completely eluted by 0.1 M Na/sub 2/HPO/sub 4/-0.5 M NaCl, pH 8. An antibody made aganist human urinary erythropoietin completely inactivated the erythropoietic factor in the dog kidney extract. Serum from a donor dog had no erythropoietin activity when assayed in the starved rat, suggesting that the factor in the extracts is intracellular erythropoietin rather than that contained in plasma trapped in the renal vasculature. The complete inactivation of the erythropoietic factor in these kidney homogenates by antierythropoietin and its behavior on DEAE-cellulose indicate that this factor is structurally similar to native plasma erythropoietin. The extracts are completely active without being incubated in the presence of serum.

• Research Organization: Univ. of Chicago
• OSTI ID: 6624019
• Journal Information: Endocrinology; (United States), Journal Name: Endocrinology; (United States) Vol. 103:3; ISSN ENDOA
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOASSAY
BIOSYNTHESIS
BODY
CARBOXYLIC ACIDS
DAYS LIVING RADIOISOTOPES
DOGS
ERYTHROPOIETIN
EVEN-ODD NUCLEI
EXTRACTION
GLOBIN
HEMOGLOBIN
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
INACTIVATION
INTERMEDIATE MASS NUCLEI
IRON 59
IRON ISOTOPES
ISOTOPES
KIDNEYS
MAMMALS
METABOLISM
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
PIGMENTS
PORPHYRINS
PROTEINS
RADIOISOTOPES
RATS
RODENTS
SEPARATION PROCESSES
SYNTHESIS
VERTEBRATES