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ERK1/2 mediates insulin stimulation of Na,K-ATPase by phosphorylation of the α-subunit in human skeletal muscle cells

Al-Khalili, Lubna ; Kotova, Olga LU ; Tsuchida, Hiroki ; Ehrén, Ingrid ; Féraille, Eric ; Krook, Anna and Chibalin, Alexander V. (2004) In Journal of Biological Chemistry 279(24). p.25211-25218
Abstract

Insulin stimulates Na+,K+-ATPase activity and induces translocation of Na+,K+-ATPase molecules to the plasma membrane in skeletal muscle. We determined the molecular mechanism by which insulin regulates Na+,K+-ATPase in differentiated primary human skeletal muscle cells (HSMCs). Insulin action on Na +,K+-ATPase was dependent on ERK1/2 in HSMCs. Sequence analysis of Na+,K+-ATPase α-subunits revealed several potential ERK phosphorylation sites. Insulin increased ouabain-sensitive 86Rb+ uptake and [3H]ouabain binding in intact cells. Insulin also increased phosphorylation and plasma membrane content... (More)

Insulin stimulates Na+,K+-ATPase activity and induces translocation of Na+,K+-ATPase molecules to the plasma membrane in skeletal muscle. We determined the molecular mechanism by which insulin regulates Na+,K+-ATPase in differentiated primary human skeletal muscle cells (HSMCs). Insulin action on Na +,K+-ATPase was dependent on ERK1/2 in HSMCs. Sequence analysis of Na+,K+-ATPase α-subunits revealed several potential ERK phosphorylation sites. Insulin increased ouabain-sensitive 86Rb+ uptake and [3H]ouabain binding in intact cells. Insulin also increased phosphorylation and plasma membrane content of the Na+,K+-ATPase α 1- and α2-subunits. Insulin-stimulated Na +,K+-ATPase activation, phosphorylation, and translocation of α-subunits to the plasma membrane were abolished by 20 μM PD98059, which is an inhibitor of MEK1/2, an upstream kinase of ERK1/2. Furthermore, inhibitors of phosphatidylinositol 3-kinase (100 wortmannin) and protein kinase C (10 μM GF109203X) had similar effects. Notably, insulin-stimulated ERK1/2 phosphorylation was abolished by wortmannin and GF109203X in HSMCS. Insulin also stimulated phosphorylation of α 1- and α2-subunits on Thr-Pro amino acid motifs, which form specific ERK substrates. Furthermore, recombinant ERK1 and -2 kinases were able to phosphorylate α-subunit of purified human Na +,K+-ATPase in vitro. In conclusion, insulin stimulates Na+,K+-ATPase activity and translocation to plasma membrane in HSMCs via phosphorylation of the α-subunits by ERK1/2 mitogen-activated protein kinase.

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author
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publishing date
type
Contribution to journal
publication status
published
in
Journal of Biological Chemistry
volume
279
issue
24
pages
8 pages
publisher
American Society for Biochemistry and Molecular Biology
external identifiers
  • scopus:2942584974
  • pmid:15069082
ISSN
0021-9258
DOI
10.1074/jbc.M402152200
language
English
LU publication?
no
id
fbe62b4a-8ba7-4a46-b21a-353c7f1844ff
date added to LUP
2019-06-03 13:18:12
date last changed
2024-01-01 09:05:38
@article{fbe62b4a-8ba7-4a46-b21a-353c7f1844ff,
  abstract     = {{<p>Insulin stimulates Na<sup>+</sup>,K<sup>+</sup>-ATPase activity and induces translocation of Na<sup>+</sup>,K<sup>+</sup>-ATPase molecules to the plasma membrane in skeletal muscle. We determined the molecular mechanism by which insulin regulates Na<sup>+</sup>,K<sup>+</sup>-ATPase in differentiated primary human skeletal muscle cells (HSMCs). Insulin action on Na <sup>+</sup>,K<sup>+</sup>-ATPase was dependent on ERK1/2 in HSMCs. Sequence analysis of Na<sup>+</sup>,K<sup>+</sup>-ATPase α-subunits revealed several potential ERK phosphorylation sites. Insulin increased ouabain-sensitive <sup>86</sup>Rb<sup>+</sup> uptake and [<sup>3</sup>H]ouabain binding in intact cells. Insulin also increased phosphorylation and plasma membrane content of the Na<sup>+</sup>,K<sup>+</sup>-ATPase α <sub>1</sub>- and α<sub>2</sub>-subunits. Insulin-stimulated Na <sup>+</sup>,K<sup>+</sup>-ATPase activation, phosphorylation, and translocation of α-subunits to the plasma membrane were abolished by 20 μM PD98059, which is an inhibitor of MEK1/2, an upstream kinase of ERK1/2. Furthermore, inhibitors of phosphatidylinositol 3-kinase (100 wortmannin) and protein kinase C (10 μM GF109203X) had similar effects. Notably, insulin-stimulated ERK1/2 phosphorylation was abolished by wortmannin and GF109203X in HSMCS. Insulin also stimulated phosphorylation of α <sub>1</sub>- and α<sub>2</sub>-subunits on Thr-Pro amino acid motifs, which form specific ERK substrates. Furthermore, recombinant ERK1 and -2 kinases were able to phosphorylate α-subunit of purified human Na <sup>+</sup>,K<sup>+</sup>-ATPase in vitro. In conclusion, insulin stimulates Na<sup>+</sup>,K<sup>+</sup>-ATPase activity and translocation to plasma membrane in HSMCs via phosphorylation of the α-subunits by ERK1/2 mitogen-activated protein kinase.</p>}},
  author       = {{Al-Khalili, Lubna and Kotova, Olga and Tsuchida, Hiroki and Ehrén, Ingrid and Féraille, Eric and Krook, Anna and Chibalin, Alexander V.}},
  issn         = {{0021-9258}},
  language     = {{eng}},
  month        = {{06}},
  number       = {{24}},
  pages        = {{25211--25218}},
  publisher    = {{American Society for Biochemistry and Molecular Biology}},
  series       = {{Journal of Biological Chemistry}},
  title        = {{ERK1/2 mediates insulin stimulation of Na,K-ATPase by phosphorylation of the α-subunit in human skeletal muscle cells}},
  url          = {{http://dx.doi.org/10.1074/jbc.M402152200}},
  doi          = {{10.1074/jbc.M402152200}},
  volume       = {{279}},
  year         = {{2004}},
}