Central role of calcium-dependent tyrosine kinase PYK2 in endothelial nitric oxide synthase-mediated angiogenic response and vascular function

Circulation. 2007 Aug 28;116(9):1041-51. doi: 10.1161/CIRCULATIONAHA.106.645416. Epub 2007 Aug 13.

Abstract

Background: The involvement of Ca2+-dependent tyrosine kinase PYK2 in the Akt/endothelial NO synthase pathway remains to be determined.

Methods and results: Blood flow recovery and neovessel formation after hind-limb ischemia were impaired in PYK2-/- mice with reduced mobilization of endothelial progenitors. Vascular endothelial growth factor (VEGF)-mediated cytoplasmic Ca2+ mobilization and Ca2+-independent Akt activation were markedly decreased in the PYK2-deficient aortic endothelial cells, whereas the Ca2+-independent AMP-activated protein kinase/protein kinase-A pathway that phosphorylates endothelial NO synthase was not impaired. Acetylcholine-mediated aortic vasorelaxation and cGMP production were significantly decreased. Vascular endothelial growth factor-dependent migration, tube formation, and actin cytoskeletal reorganization associated with Rac1 activation were inhibited in PYK2-deficient endothelial cells. PI3-kinase is associated with vascular endothelial growth factor-induced PYK2/Src complex, and inhibition of Src blocked Akt activation. The vascular endothelial growth factor-mediated Src association with PLCgamma1 and phosphorylation of 783Tyr-PLCgamma1 also were abolished by PYK2 deficiency.

Conclusion: These findings demonstrate that PYK2 is closely involved in receptor- or ischemia-activated signaling events via Src/PLCgamma1 and Src/PI3-kinase/Akt pathways, leading to endothelial NO synthase phosphorylation, and thus modulates endothelial NO synthase-mediated vasoactive function and angiogenic response.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analysis of Variance
  • Animals
  • Calcium / physiology
  • Enzyme Activation
  • Focal Adhesion Kinase 2 / deficiency
  • Focal Adhesion Kinase 2 / physiology*
  • Heart / physiology*
  • Hindlimb / blood supply
  • Ischemia / physiopathology
  • Mice
  • Mice, Knockout
  • Neovascularization, Physiologic / physiology*
  • Nitric Oxide Synthase Type III / metabolism*
  • Oncogene Protein v-akt / physiology*
  • Phosphorylation
  • Signal Transduction
  • Vascular Endothelial Growth Factor A / physiology
  • Vasodilation

Substances

  • Vascular Endothelial Growth Factor A
  • Nitric Oxide Synthase Type III
  • Focal Adhesion Kinase 2
  • Oncogene Protein v-akt
  • Calcium