Na+,K+-ATPase Na+ affinity in rat skeletal muscle fiber types.

Previous studies in expression systems have found different ion activation of the Na(+)/K(+)-ATPase isozymes, which suggest that different muscles have different ion affinities. The rate of ATP hydrolysis was used to quantify Na(+),K(+)-ATPase activity, and the Na(+) affinity of Na(+),K(+)-ATPase was studied in total membranes from rat muscle and purified membranes from muscle with different fiber types. The Na(+) affinity was higher (K(m) lower) in oxidative muscle compared with glycolytic muscle and in purified membranes from oxidative muscle compared with glycolytic muscle. Na(+),K(+)-ATPase isoform analysis implied that heterodimers containing the beta(1) isoform have a higher Na(+) affinity than heterodimers containing the beta(2) isoform. Immunoprecipitation experiments demonstrated that dimers with alpha(1) are responsible for approximately 36% of the total Na,K-ATPase activity. Selective inhibition of the alpha(2) isoform with ouabain suggested that heterodimers containing the alpha(1) isoform have a higher Na(+) affinity than heterodimers containing the alpha(2) isoform. The estimated K(m) values for Na(+) are 4.0, 5.5, 7.5 and 13 mM for alpha(1)beta(1), alpha(2)beta(1), alpha(1)beta(2) and alpha(2)beta(2), respectively. The affinity differences and isoform distributions imply that the degree of activation of Na(+),K(+)-ATPase at physiological Na(+) concentrations differs between muscles (oxidative and glycolytic) and between subcellular membrane domains with different isoform compositions. These differences may have consequences for ion balance across the muscle membrane.