Central role of calcium-dependent tyrosine kinase PYK2 in endothelial nitric oxide synthase-mediated angiogenic response and vascular function.
BACKGROUND
The involvement of Ca2+-dependent tyrosine kinase PYK2 in the Akt/endothelial NO synthase pathway remains to be determined.


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.


CONCLUSIONS
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.
Matsui
Akihiro
A
Department of Cardiovascular Medicine, Kyoto Prefectural University of Medicine, Kamigyo-ku, Kyoto, 602-8566, Japan.
Okigaki
Mitsuhiko
M
Amano
Katsuya
K
Adachi
Yasushi
Y
Jin
Denan
D
Takai
Shinji
S
Yamashita
Tomoya
T
Kawashima
Seinosuke
S
Kurihara
Tatsuya
T
Miyazaki
Mizuo
M
Tateishi
Kento
K
Matsunaga
Shinsaku
S
Katsume
Asako
A
Honshou
Shoken
S
Takahashi
Tomosaburo
T
Matoba
Satoaki
S
Kusaba
Tetsuro
T
Tatsumi
Tetsuya
T
Matsubara
Hiroaki
H
D000704:Analysis of Variance; D000818:Animals; D002118:Calcium / Q000502:physiology; D004789:Enzyme Activation; D051418:Focal Adhesion Kinase 2 / Q000172:deficiency / Q000502:physiology*; D006321:Heart / Q000502:physiology*; D006614:Hindlimb / Q000098:blood supply; D007511:Ischemia / Q000503:physiopathology; D051379:Mice; D018345:Mice, Knockout; D018919:Neovascularization, Physiologic / Q000502:physiology*; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism*; D051058:Oncogene Protein v-akt / Q000502:physiology*; D010766:Phosphorylation; D015398:Signal Transduction; D042461:Vascular Endothelial Growth Factor A / Q000502:physiology; D014664:Vasodilation
D042461:Vascular Endothelial Growth Factor A; D052250:Nitric Oxide Synthase Type III; D051418:Focal Adhesion Kinase 2; D051058:Oncogene Protein v-akt; D002118:Calcium