Antisera specific for the .alpha.1, .alpha.2, .alpha.3, and .beta. subunits of the sodium-potassium ATPase: differential expression of .alpha. and .beta. subunits in rat tissue membranes

Abstract

We have developed a panel of antibodies specific for the alpha 1, alpha 2, alpha 3, and beta subunits of the rat Na,K-ATPase. TrpE-alpha subunit isoform fusion proteins were used to generate three antisera, each of which reacted specifically with a distinct alpha subunit isotype. Western blot analysis of rat tissue microsomes revealed that alpha 1 subunits were expressed in all tissues while alpha 2 subunits were expressed in brain, heart, and lung. The alpha 3 subunit, a protein whose existence had been inferred from cDNA cloning, was expressed primarily in brain and copurified with ouabain-inhibitable Na,K-ATPase activity. An antiserum specific for the rat Na,K-ATPase beta subunit was generated from a TrpE-beta subunit fusion protein. Western blot analysis showed that beta subunits were present in kidney, brain, and heart. However, no beta subunits were detected in liver, lung, spleen, thymus, or lactating mammary gland. The distinct tissue distributions of alpha and beta subunits suggest that different members of the Na,K-ATPase family may have specialized functions.

α3Na+/K+-ATPase Deficiency Causes Brain Ventricle Dilation and Abrupt Embryonic Motility in Zebrafish

“…The Atp1a3a transcript was also abundant in the spinal cord and, to a lesser extent, in the heart. The latter observation is consistent with previous studies demonstrating ATP1␣ 3 expression in the neonatal, but not adult, rat heart (40,41).…”

Decreased hepatocyte membrane potential differences and GABAa-β3 expression in human hepatocellular carcinoma

“…Although there is general agreement on the presence of the ␣1 subunit, the status of the ␤1 subunit is less clear. 13,35,36 We were able to detect both ␣1 and ␤1 expression via RT-PCR. However, the pattern of each subunit expression was similar (and certainly not decreased, which would be required to explain HCC depolarization relative to adjacent nontumor tissues).…”

Subunit Composition and Role of Na+,K+-ATPases in Ventricular Myocytes

“…The antibodies used were a monoclonal anti-Na,K-ATPase α1 subunit antibody (mAb) (05-369; Upstate Biotechnology) [12], rabbit anti-Na,K-ATPase α2 Ab (06-168; Upstate) [13], anti-Na,K-ATPase α3 mAb (MA3-915; Affnity BioReagents) [14], rabbit antiNa,K-ATPase α3 Ab (06-172; Upstate) [13], anti-Na,KATPase β1 mAb (05-382; Upstate) [15], rabbit antiNa,K-ATPase β1 Ab (06-170; Upstate) [13], anti-Na,KATPase β2 mAb (610914; BD Transduction Laboratories) [16], rabbit anti-Na,K-ATPase β2 Ab (06-171; Upstate) [17], rabbit anti-Na,K-ATPase β3 Ab (06-817; Upstate) [7], rabbit anti-SERCA pump mAb (MA3-910; Affinity BioReagents) [18], and anti-N-cadherin mAb (610920; BD) [19].…”