Differential distribution and regulation of mouse cardiac Na+/K+-ATPase α1 and α2 subunits in T-tubule and surface sarcolemmal membranes

Roger G. Berry, Sanda Despa, William Fuller, Donald M Bers, Michael J. Shattock

Research output: Contribution to journalArticle

71 Scopus citations

Abstract

Objectives: Two Na+/K+-ATPase (NKA) α-subunit isoforms, α1 and α2, are expressed in the adult mouse heart. The subcellular distribution of these isoforms in T-tubule and surface sarcolemmal (SSL) membranes and their regulation by cAMP-dependent protein kinase (PKA) is unclear. Methods: We used formamide-induced detubulation of mouse ventricular myocytes to investigate differential functional distribution and regulation by PKA of α1 and α2 in T-tubule versus SSL membranes by measuring NKA current (Ipump) and NKA-mediated Na+ efflux (- d[Na]i/dt). Results: Ipump is composed of 88% α1-mediated Ipump (Iα1) and 12% α2-mediated Ipump (Iα2). α1 and α2 subunits demonstrate distinct ouabain affinities (105 ± 6 and 0.3 ± 0.1 μmol/L respectively) but similar affinity for intracellular Na+ (K1/2Na+ of 16.6 ± 0.8 and 16.7±2.6 mmol/L respectively). Detubulation reduced (i) Ipump density (1.42 ± 0.1 to 1.20 ± 0.04 pA/pF), (ii) cell capacitance (181 ± 12 to 127±17 pF), and (iii) Iα2 contribution (12 to 6%). Total Ipump density was ∼ 60% higher in T-tubule (1.94 pA/pF, derived) vs. SSL membranes. Although T-tubule membranes represent only 30% of total surface area, they generate ∼ 70% of Iα2 and ∼ 37% of Iα1. Iα1 density was substantially higher than Iα2 in SSL (Iα1:Iα2 = 16:1) but this was markedly reduced in T-tubules (4:1). In addition to differential localisation, isoprenaline (ISO, 1 μmol/L) significantly increased α1-mediated NKA Na+ affinity (from 16.6 ± 0.8 to 13.3 ± 1.4 mmol/L) and caused a small increase in maximal NKA Na+ efflux rate. ISO had no effect on α2-mediated NKA activity. Conclusion: These data suggest that NKA α1 and α2 subunits are differentially localised and regulated by PKA in T-tubule and SSL membranes and may have distinct regulatory roles in cardiac excitation-contraction coupling.

Original languageEnglish (US)
Pages (from-to)92-100
Number of pages9
JournalCardiovascular Research
Volume73
Issue number1
DOIs
StatePublished - Jan 1 2007
Externally publishedYes

Keywords

  • Adrenergic agonist
  • Ion pumps
  • Na/K-pump
  • Protein kinase A
  • Sarcolemmal

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

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