Extracellular potassium dependence of the Na+-K+-ATPase in cardiac myocytes: isoform specificity and effect of phospholemman

Abstract

Cardiac Na+-K+-ATPase (NKA) regulates intracellular Na+, which in turn affects intracellular Ca2+and contractility via the Na+/Ca2+exchanger. Extracellular K+concentration ([K+]) is a central regulator of NKA activity. Phospholemman (PLM) has recently been recognized as a critical regulator of NKA in the heart. PLM reduces the intracellular Na+affinity of NKA, an effect relieved by PLM phosphorylation. Here we tested whether the NKA α1- vs. α2- isoforms have different external K+sensitivity and whether PLM and PKA activation affects the NKA affinity for K+in mouse cardiac myocytes. We measured the external [K+] dependence of the pump current generated by the ouabain-resistant NKA isoform in myocytes from wild-type (WT) mice (i.e., current due to NKA-α1) and mice in which the NKA isoforms have swapped ouabain affinities (α1is ouabain sensitive and α2is ouabain resistant) to assess current due to NKA-α2. We found that NKA-α1has a higher affinity for external K+than NKA-α2[half-maximal pump activation ( K0.5) = 1.5 ± 0.1 vs. 2.9 ± 0.3 mM]. The apparent external K+affinity of NKA was significantly lower in myocytes from WT vs. PLM-knockout mice ( K0.5= 2.0 ± 0.2 vs. 1.05 ± 0.08 mM). However, PKA activation by isoproterenol (1 μM) did not alter the K0.5of NKA for external K+in WT myocytes. We conclude that 1) NKA-α1has higher affinity for K+than NKA-α2in cardiac myocytes, 2) PLM decreases the apparent external K+affinity of NKA, and 3) phosphorylation of PLM at the cytosolic domain does not alter apparent extracellular K+affinity of NKA.