The (Na+, K+)ATPase of rat kidney: purification, biosynthesis, and processing.
(Na+, K+)ATPase was purified from rat renal outer medulla by concanavalin A- and wheat germ agglutinin-lectin Sepharose affinity chromatographies. The antibody, which was raised in rabbits, markedly inhibited ATPase activity. The monospecificity of this antibody was assayed by the Ouchterlony double immunodiffusion and Western blotting tests. The endoplasmic reticulum (ER)-rich, and Golgi-rich subfractions were prepared from the rat kidney microsomal fraction by sucrose density gradient centrifugation. On the immunoblot, the molecular weight of the alpha subunit in both fractions was 95 kilodalton (Kd); whereas, that of the beta subunit was 50 Kd in the ER-rich fraction and 54 Kd in the Golgi-rich fraction. When treated with endoglucosidase H, the 50 Kd component was converted to 38 Kd, but the 54 Kd component was endoglucosidase H resistant. These results suggest that the beta subunit (38 Kd) is glycosylated cotranslationally in the ER (50 Kd) then is converted to the mature type subunit (54 Kd) in the Golgi apparatus.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D002458:Cell Fractionation; D002846:Chromatography, Affinity; D004721:Endoplasmic Reticulum / Q000201:enzymology / Q000378:metabolism; D006056:Golgi Apparatus / Q000201:enzymology; D007668:Kidney / Q000201:enzymology*; D007679:Kidney Medulla / Q000166:cytology; D008297:Male; D008861:Microsomes / Q000201:enzymology; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains; D000254:Sodium-Potassium-Exchanging ATPase / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism
D000596:Amino Acids; D000906:Antibodies; D000254:Sodium-Potassium-Exchanging ATPase