title: Characterization of erythropoietin isolated from rat serum: biochemical comparison of rat and human erythropoietins.
abstract: We isolated erythropoietin (Epo) from anemic-rat serum with 1.3 x 10(6)-fold purification and 38% recovery using immunoaffinity chromatography. The isolated Epo migrated in SDS polyacrylamide gel with a molecular size of 37 kDa. Biological properties of rat Epo were compared with those of human Epo using target cells of primate and murine origins. When murine cells were used as target cells for assaying Epo, rat Epo stimulated proliferation of the cells with a 50% lower potency than did human Epo. The activity of rat Epo on human cells was only 25% of that of human Epo. Studies of Epo binding to the receptor indicated that rat and human Epos were not distinguishable in binding to murine cells; however, rat Epo bound to the receptor on human cells with an affinity much lower than that of human Epo. Rat Epo was digested with N-glycanase. Complete removal of N-linked sugars converted the native Epo to the deglycosylated form with 18 kDa. The in vitro activity of deglycosylated Epo was 2.5-fold higher than that of the native Epo.
authors.affiliation: Department of Food Science and Technology, Faculty of Agriculture, Kyoto University, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Okano
authors.forename: H
authors.initials: H
authors.lastname: Suga
authors.forename: S
authors.initials: S
authors.lastname: Masuda
authors.forename: M
authors.initials: M
authors.lastname: Nagao
authors.forename: H
authors.initials: H
authors.lastname: Narita
authors.forename: K
authors.initials: K
authors.lastname: Ikura
authors.forename: R
authors.initials: R
authors.lastname: Sasaki
mesh_terms: D000595:Amino Acid Sequence; D000740:Anemia / Q000097:blood; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D004921:Erythropoietin / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D051381:Rats; D017467:Receptors, Erythropoietin / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology
chemical_list: D017467:Receptors, Erythropoietin; D011994:Recombinant Proteins; D004921:Erythropoietin