AMP-dependent kinase inhibits oxidative stress-induced caveolin-1 phosphorylation and endocytosis by suppressing the dissociation between c-Abl and Prdx1 proteins in endothelial cells

Kimio Takeuchi, Yuki Morizane, Cynthia Kamami-Levy, Jun Suzuki, Maki Kayama, Wenyi Cai, Joan W. Miller, Demetrios G. Vavvas

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Caveolin-1 is the primary structural component of endothelial caveolae that is essential for transcellular trafficking of albumin and is also a critical scaffolding protein that regulates the activity of signaling molecules in caveolae. Phosphorylation of caveolin-1 plays a fundamental role in the mechanism of oxidant-induced vascular hyper permeability. However, the regulatory mechanism of caveolin-1 phosphorylation remains unclear. Here we identify a previously unexpected role for AMPKin inhibition of caveolin-1 phosphorylation under oxidative stress. A pharmacological activator of AMPK, 5-amino-4-imidazole carboxamide riboside (AICAR), inhibited oxidative stress-induced phosphorylation of both caveolin-1 and c-Abl, which is the major kinase of caveolin-1, and endocytosis of albumin in human umbilical vein endothelial cell. These effects were abolished by treatment with two specific inhibitors of AICAR, dipyridamole, and 5-iodotubericidin. Consistently, knockdown of the catalytic AMPKα subunit by siRNA abolished the inhibitory effect of AICAR on oxidant-induced phosphorylation of both caveolin-1 and c-Abl. Pretreatment with specific c-Abl inhibitor, imatinib mesylate, and knock down of c-Abl significantly decreased the caveolin-1 phosphorylation after H2O2 exposure and abolished the inhibitory effect of AICAR on the caveolin-1 phosphorylation. Interestingly, knockdown of Prdx-1, an antioxidant enzyme associated with c-Abl, increased phosphorylation of both caveolin-1 and c-Abl and abolished the inhibitory effect of AICAR on the caveolin-1 phosphorylation. Furthermore, co-immunoprecipitation experiment showed that AICAR suppressed the oxidant-induced dissociation between c-Abl and Prdx1. Overall, our results suggest that activation of AMPK inhibits oxidative stress-induced caveolin-1 phosphorylation and endocytosis, and this effect is mediated in part by stabilizing the interaction between c-Abl and Prdx-1.

Original languageEnglish
Pages (from-to)20581-20591
Number of pages11
JournalJournal of Biological Chemistry
Volume288
Issue number28
DOIs
Publication statusPublished - Jul 12 2013

Fingerprint

Proto-Oncogene Proteins c-abl
Caveolin 1
Adenylate Kinase
Phosphorylation
Oxidative stress
Endothelial cells
Adenosine Monophosphate
Endocytosis
Oxidative Stress
Phosphotransferases
Endothelial Cells
Proteins
AMP-Activated Protein Kinases
Oxidants
Caveolae
Albumins
Dipyridamole
Human Umbilical Vein Endothelial Cells
Capillary Permeability

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

AMP-dependent kinase inhibits oxidative stress-induced caveolin-1 phosphorylation and endocytosis by suppressing the dissociation between c-Abl and Prdx1 proteins in endothelial cells. / Takeuchi, Kimio; Morizane, Yuki; Kamami-Levy, Cynthia; Suzuki, Jun; Kayama, Maki; Cai, Wenyi; Miller, Joan W.; Vavvas, Demetrios G.

In: Journal of Biological Chemistry, Vol. 288, No. 28, 12.07.2013, p. 20581-20591.

Research output: Contribution to journalArticle

Takeuchi, Kimio ; Morizane, Yuki ; Kamami-Levy, Cynthia ; Suzuki, Jun ; Kayama, Maki ; Cai, Wenyi ; Miller, Joan W. ; Vavvas, Demetrios G. / AMP-dependent kinase inhibits oxidative stress-induced caveolin-1 phosphorylation and endocytosis by suppressing the dissociation between c-Abl and Prdx1 proteins in endothelial cells. In: Journal of Biological Chemistry. 2013 ; Vol. 288, No. 28. pp. 20581-20591.
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