N-terminal domain of soluble epoxide hydrolase negatively regulates the VEGF-mediated activation of endothelial nitric oxide synthase

Hsin Han Hou, Bruce D. Hammock, Kou Hui Su, Christophe Morisseau, Yu Ru Kou, Susumu Imaoka, Ami Oguro, Song Kun Shyue, Jin Feng Zhao, Tzong Shyuan Lee

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Aims The mammalian soluble epoxide hydrolase (sEH) has both an epoxide hydrolase and a phosphatase domain. The role of sEH hydrolase activity in the metabolism of epoxyeicosatrienoic acids (EETs) and the activation of endothelial nitric oxide synthase (eNOS) in endothelial cells (ECs) has been well defined. However, far less is known about the role of sEH phosphatase activity in eNOS activation. In the present study, we investigated whether the phosphatase domain of sEH was involved in the eNOS activation in ECs. Methods and Results The level of eNOS phosphorylation in aortas is higher in the sEH knockout (sEH -/-) mice than in wild-type mice. In ECs, pharmacological inhibition of sEH phosphatase or overexpressing sEH with an inactive phosphatase domain enhanced vascular endothelial growth factor (VEGF)-induced NO production and eNOS phosphorylation. In contrast, overexpressing the phosphatase domain of sEH prevented the VEGF-mediated NO production and eNOS phosphorylation at Ser617, Ser635, and Ser1179. Additionally, treatment with VEGF induced a c-Src kinase-dependent increase in transient tyrosine phosphorylation of sEH and the formation of a sEHeNOS complex, which was abolished by treatment with a c-Src kinase inhibitor, PP1, or the c-Src dominant-negative mutant K298M. We also demonstrated that the phosphatase domain of sEH played a key role in VEGF-induced angiogenesis by detecting the tube formation in ECs and neovascularization in Matrigel plugs in mice. Conclusion In addition to epoxide hydrolase activity, phosphatase activity of sEH plays a pivotal role in the regulation of eNOS activity and NO-mediated EC functions.

Original languageEnglish (US)
Pages (from-to)120-129
Number of pages10
JournalCardiovascular Research
Volume93
Issue number1
DOIs
StatePublished - Jan 1 2012

Fingerprint

Epoxide Hydrolases
Nitric Oxide Synthase Type III
Vascular Endothelial Growth Factor A
Phosphoric Monoester Hydrolases
Endothelial Cells
Phosphorylation
Hydrolases
Knockout Mice
Tyrosine
Aorta

Keywords

  • Angiogenesis
  • c-Src kinase
  • Endothelial nitric oxide synthase
  • Phosphatase
  • Soluble epoxide hydrolase

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)
  • Physiology

Cite this

N-terminal domain of soluble epoxide hydrolase negatively regulates the VEGF-mediated activation of endothelial nitric oxide synthase. / Hou, Hsin Han; Hammock, Bruce D.; Su, Kou Hui; Morisseau, Christophe; Kou, Yu Ru; Imaoka, Susumu; Oguro, Ami; Shyue, Song Kun; Zhao, Jin Feng; Lee, Tzong Shyuan.

In: Cardiovascular Research, Vol. 93, No. 1, 01.01.2012, p. 120-129.

Research output: Contribution to journalArticle

Hou, HH, Hammock, BD, Su, KH, Morisseau, C, Kou, YR, Imaoka, S, Oguro, A, Shyue, SK, Zhao, JF & Lee, TS 2012, 'N-terminal domain of soluble epoxide hydrolase negatively regulates the VEGF-mediated activation of endothelial nitric oxide synthase', Cardiovascular Research, vol. 93, no. 1, pp. 120-129. https://doi.org/10.1093/cvr/cvr267
Hou, Hsin Han ; Hammock, Bruce D. ; Su, Kou Hui ; Morisseau, Christophe ; Kou, Yu Ru ; Imaoka, Susumu ; Oguro, Ami ; Shyue, Song Kun ; Zhao, Jin Feng ; Lee, Tzong Shyuan. / N-terminal domain of soluble epoxide hydrolase negatively regulates the VEGF-mediated activation of endothelial nitric oxide synthase. In: Cardiovascular Research. 2012 ; Vol. 93, No. 1. pp. 120-129.
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abstract = "Aims The mammalian soluble epoxide hydrolase (sEH) has both an epoxide hydrolase and a phosphatase domain. The role of sEH hydrolase activity in the metabolism of epoxyeicosatrienoic acids (EETs) and the activation of endothelial nitric oxide synthase (eNOS) in endothelial cells (ECs) has been well defined. However, far less is known about the role of sEH phosphatase activity in eNOS activation. In the present study, we investigated whether the phosphatase domain of sEH was involved in the eNOS activation in ECs. Methods and Results The level of eNOS phosphorylation in aortas is higher in the sEH knockout (sEH -/-) mice than in wild-type mice. In ECs, pharmacological inhibition of sEH phosphatase or overexpressing sEH with an inactive phosphatase domain enhanced vascular endothelial growth factor (VEGF)-induced NO production and eNOS phosphorylation. In contrast, overexpressing the phosphatase domain of sEH prevented the VEGF-mediated NO production and eNOS phosphorylation at Ser617, Ser635, and Ser1179. Additionally, treatment with VEGF induced a c-Src kinase-dependent increase in transient tyrosine phosphorylation of sEH and the formation of a sEHeNOS complex, which was abolished by treatment with a c-Src kinase inhibitor, PP1, or the c-Src dominant-negative mutant K298M. We also demonstrated that the phosphatase domain of sEH played a key role in VEGF-induced angiogenesis by detecting the tube formation in ECs and neovascularization in Matrigel plugs in mice. Conclusion In addition to epoxide hydrolase activity, phosphatase activity of sEH plays a pivotal role in the regulation of eNOS activity and NO-mediated EC functions.",
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T1 - N-terminal domain of soluble epoxide hydrolase negatively regulates the VEGF-mediated activation of endothelial nitric oxide synthase

AU - Hou, Hsin Han

AU - Hammock, Bruce D.

AU - Su, Kou Hui

AU - Morisseau, Christophe

AU - Kou, Yu Ru

AU - Imaoka, Susumu

AU - Oguro, Ami

AU - Shyue, Song Kun

AU - Zhao, Jin Feng

AU - Lee, Tzong Shyuan

PY - 2012/1/1

Y1 - 2012/1/1

N2 - Aims The mammalian soluble epoxide hydrolase (sEH) has both an epoxide hydrolase and a phosphatase domain. The role of sEH hydrolase activity in the metabolism of epoxyeicosatrienoic acids (EETs) and the activation of endothelial nitric oxide synthase (eNOS) in endothelial cells (ECs) has been well defined. However, far less is known about the role of sEH phosphatase activity in eNOS activation. In the present study, we investigated whether the phosphatase domain of sEH was involved in the eNOS activation in ECs. Methods and Results The level of eNOS phosphorylation in aortas is higher in the sEH knockout (sEH -/-) mice than in wild-type mice. In ECs, pharmacological inhibition of sEH phosphatase or overexpressing sEH with an inactive phosphatase domain enhanced vascular endothelial growth factor (VEGF)-induced NO production and eNOS phosphorylation. In contrast, overexpressing the phosphatase domain of sEH prevented the VEGF-mediated NO production and eNOS phosphorylation at Ser617, Ser635, and Ser1179. Additionally, treatment with VEGF induced a c-Src kinase-dependent increase in transient tyrosine phosphorylation of sEH and the formation of a sEHeNOS complex, which was abolished by treatment with a c-Src kinase inhibitor, PP1, or the c-Src dominant-negative mutant K298M. We also demonstrated that the phosphatase domain of sEH played a key role in VEGF-induced angiogenesis by detecting the tube formation in ECs and neovascularization in Matrigel plugs in mice. Conclusion In addition to epoxide hydrolase activity, phosphatase activity of sEH plays a pivotal role in the regulation of eNOS activity and NO-mediated EC functions.

AB - Aims The mammalian soluble epoxide hydrolase (sEH) has both an epoxide hydrolase and a phosphatase domain. The role of sEH hydrolase activity in the metabolism of epoxyeicosatrienoic acids (EETs) and the activation of endothelial nitric oxide synthase (eNOS) in endothelial cells (ECs) has been well defined. However, far less is known about the role of sEH phosphatase activity in eNOS activation. In the present study, we investigated whether the phosphatase domain of sEH was involved in the eNOS activation in ECs. Methods and Results The level of eNOS phosphorylation in aortas is higher in the sEH knockout (sEH -/-) mice than in wild-type mice. In ECs, pharmacological inhibition of sEH phosphatase or overexpressing sEH with an inactive phosphatase domain enhanced vascular endothelial growth factor (VEGF)-induced NO production and eNOS phosphorylation. In contrast, overexpressing the phosphatase domain of sEH prevented the VEGF-mediated NO production and eNOS phosphorylation at Ser617, Ser635, and Ser1179. Additionally, treatment with VEGF induced a c-Src kinase-dependent increase in transient tyrosine phosphorylation of sEH and the formation of a sEHeNOS complex, which was abolished by treatment with a c-Src kinase inhibitor, PP1, or the c-Src dominant-negative mutant K298M. We also demonstrated that the phosphatase domain of sEH played a key role in VEGF-induced angiogenesis by detecting the tube formation in ECs and neovascularization in Matrigel plugs in mice. Conclusion In addition to epoxide hydrolase activity, phosphatase activity of sEH plays a pivotal role in the regulation of eNOS activity and NO-mediated EC functions.

KW - Angiogenesis

KW - c-Src kinase

KW - Endothelial nitric oxide synthase

KW - Phosphatase

KW - Soluble epoxide hydrolase

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