Role of phospholemman phosphorylation sites in mediating kinase-dependent regulation of the Na+-K+-ATPase

Fei Han, Julie Bossuyt, Jody L. Martin, Sanda Despa, Donald M. Bers

Research output: Contribution to journalArticlepeer-review

29 Scopus citations


Phospholemman (PLM) is a major target for phosphorylation mediated by both PKA (at Ser68) and PKC (at both Ser63 and Ser68) in the heart. In intact cardiac myocytes, PLM associates with and inhibits Na+-K+-ATPase (NKA), mainly by reducing its affinity for internal Na+. The inhibition is relieved upon PLM phosphorylation by PKA or PKC. The aim here was to distinguish the role of the Ser63 and Ser68 PLM phosphorylation sites in mediating kinase-induced modulation of NKA function. We expressed wild-type (WT) PLM and S63A, S68A, and AA (Ser63 and Ser68 to alanine double mutant) PLM mutants in HeLa cells that stably express rat NKA-α1 and we measured the effect of PKA and PKC activation on NKA-mediated intracellular Na+ concentration decline. PLM expression (WT or mutant) significantly decreased the apparent NKA affinity for internal Na+ and had no significant effect on the maximum pump rate (Vmax). PKA activation with forskolin (20 μM) restored NKA Na+ affinity in cells expressing WT but not AA PLM and did not affect Vmax in either case. Similarly, PKC activation with 300 nM phorbol 12,13-dibutyrate increased NKA Na+ affinity in cells expressing WT, S63A, and S68A PLM and had no effect in cells expressing AA PLM. Neither forskolin nor phorbol 12,13-dibutyrate affected NKA function in the absence of PLM. We conclude that PLM phosphorylation at either Ser63 or Ser68 is both necessary and sufficient for completely relieving the PLM-induced NKA inhibition.

Original languageEnglish
Pages (from-to)C1363-C1369
JournalAmerican Journal of Physiology - Cell Physiology
Issue number6
StatePublished - Dec 2010


  • Apparent Na affinity
  • FXYD
  • PKA
  • PKC

ASJC Scopus subject areas

  • Physiology
  • Cell Biology


Dive into the research topics of 'Role of phospholemman phosphorylation sites in mediating kinase-dependent regulation of the Na+-K+-ATPase'. Together they form a unique fingerprint.

Cite this