Comparison of subunits of cardiac, brain, and kidney Na+-K+-ATPase

Abstract

Na+-K+-ATPase is in low abundance in cardiac tissue. Therefore, we utilized antibodies to detect the cardiac Na+-K+-ATPase subunits and to compare their characteristics with those of kidney and brain Na+-K+-ATPase subunits. By using crude preparations of heart membranes as well as purified sarcolemmal membranes from guinea pig hearts, we resolved peptides by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, blotted them onto diazotized paper, and detected Na+-K+-ATPase subunits with antibodies generated against highly purified kidney Na+-K+-ATPase holoenzyme. We tested the hypothesis that the two families of ouabain-binding affinities described in heart are due to two forms of alpha-subunit, analogous to the two forms with different affinities for ouabain described in brain. Although the antibodies did detect two forms of catalytic subunit in brain (alpha and alpha +), only one form of alpha was detected in the heart membranes, with the same electrophoretic mobility as kidney alpha. Cardiac beta-subunit could also be detected with the antikidney antibodies. It had a similar electrophoretic mobility to that described for kidney beta, whereas brain beta had a higher mobility.