2024-03-29T14:56:58Zhttps://eprints.lib.hokudai.ac.jp/dspace-oai/requestoai:eprints.lib.hokudai.ac.jp:2115/733282023-01-19T03:10:04Zhdl_2115_20048hdl_2115_140Na,K-ATPase in dog red cells. Immunological identification and maturation-associated degradation by the proteolytic system.Inaba, MutsumiMaede, Yoshimitsu649AnimalsChromatography, AffinityCross ReactionsDogsElectrophoresis, Polyacrylamide GelErythrocytes/enzymologyHematopoietic Stem Cells/enzymologyImmunoglobulin G/immunologyMolecular WeightPeptide Hydrolases/metabolismReticulocytes/enzymologySodium-Potassium-Exchanging ATPase/bloodSodium-Potassium-Exchanging ATPase/immunologyThe Na,K-ATPase of red cells from high K+ and low K+ dogs was studied immunologically by using antibodies raised against dog kidney enzyme. Anti-alpha subunit IgGs, which also recognized alpha (+) from brain enzyme, identified the larger subunit of erythrocyte Na,K-ATPase as a homogeneous polypeptide with Mr = 96,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by immunoblotting. In addition, erythrocyte Na,K-ATPase, purified by immunoaffinity chromatography on a monoclonal antibody-coupled column, showed the identity of its polypeptide composition to that of the renal enzyme. Furthermore, it was shown that reticulocyte lysates from high K+ and low K+ dogs substantially degraded 125I-Bolton-Hunter reagent-labeled Na,K-ATPase. This degradation of the enzyme protein was significantly enhanced by the addition of ATP and Mg2+. These results indicate that dog reticulocytes possess some mechanism for protein breakdown involving an ATP-dependent proteolytic system, resulting in the dramatic breakdown of Na,K-ATPase activity during dog reticulocyte maturation into erythrocytes (Maede, Y., and Inaba, M. (1985) J. Biol. Chem. 260, 3337-3343).American Society for Biochemistry and Molecular BiologyJournal Articleapplication/pdfhttp://hdl.handle.net/2115/73328https://eprints.lib.hokudai.ac.jp/dspace/bitstream/2115/73328/1/J.%20Biol.%20Chem.261_34_16099.pdf0021-92581083-351XAA00251083The Journal of biological chemistry2613416099161051986-12-05enginfo:pmid/3023340publisher