Tissue-specific expression of the Na,K-ATPase beta3 subunit. The presence of beta3 in lung and liver addresses the problem of the missing subunit.
The Na,K-ATPase belongs to a family of P-type ion-translocating ATPases sharing homologous catalytic subunits (alpha) that traverse the membrane several times and contain the binding sites for ATP and cations. In this family, only Na,K- and H,K-ATPases have been shown to have a second subunit, a single-span glycoprotein called beta. Recently a new isoform (beta3) has been identified in mammals. Here we describe structural features and tissue distribution of the beta3 protein, utilizing an antiserum specific for its N terminus. beta3 was the only beta detected in Na,K-ATPase purified from C6 glioma. Treatment with N-glycosidase F confirmed that beta3 is a glycoprotein containing N-linked carbohydrate chains. Molecular masses of the glycosylated protein and core protein were estimated to be 42 and 35 kDa, respectively, which are different from those of the beta1 and beta2 subunits. Detection of beta subunits has historically been difficult in certain tissues. Sensitivity was improved by deglycosylating, and expression was evaluated by obtaining estimates of beta3/alpha ratio. The proportion of beta3 protein in the rat was highest in lung and testis. It was also present in liver and skeletal muscle, whereas kidney, heart, and brain contained it only as a minor component of the Na,K-ATPase. In P7 rat, we found skeletal muscle and lung Na,K-ATPase to be the most enriched in beta3 subunit, whereas expression in liver was very low, illustrating developmentally regulated changes in expression. The substantial expression in lung and adult liver very likely explains long-standing puzzles about an apparent paucity of beta subunit in membranes or in discrete cellular or subcellular structures.
Arystarkhova
E
E
Laboratory of Membrane Biology, Neuroscience Center, Massachusetts General Hospital, Charlestown, Massachusetts 02129, USA.
Sweadner
K J
KJ
D000818:Animals; D000906:Antibodies / Q000235:genetics; D006801:Humans; D008970:Molecular Weight; D017386:Sequence Homology, Amino Acid; D000254:Sodium-Potassium-Exchanging ATPase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D013045:Species Specificity; D014407:Tumor Cells, Cultured
D000906:Antibodies; D000254:Sodium-Potassium-Exchanging ATPase