Phospholipase D activation by norepinephrine is mediated by 12(S)-, 15(S)-, and 20-hydroxyeicosatetraenoic acids generated by stimulation of cytosolic phospholipase A2

Tyrosine phosphorylation of phospholipase D 2 in response to norepinephrine

Jean Hugues Parmentier, Mubarack M. Muthalif, Abdelwahab E. Saeed, Kafait Malik

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Abstract

Norepinephrine (NE) stimulates phospholipase D (PLD) through a Ras/MAPK pathway in rabbit vascular smooth muscle cells (VSMC). NE also activates calcium influx and calmodulin (CaM)-dependent protein kinase II-dependent cytosolic phospholipase A2 (cPLA2). Arachidonic acid (AA) released by cPLA2-catalyzed phospholipid hydrolysis is then metabolized into hydroxyeicosatetraenoic acids (HETEs) through lipoxygenase and cytochrome P450 4A (CYP4A) pathways. HETEs, in turn, have been shown to stimulate Ras translocation and to increase MAPK activity in VSMC. This study was conducted to determine the contribution of cPLA2-derived AA and its metabolites (HETEs) to the activation of PLD. NE-induced PLD activation was reduced by two structurally distinct CaM antagonists, W-7 and calmidazolium, and by CaM-dependent protein kinase II inhibition. Blockade of cPLA2 activity or protein depletion with selective cPLA2 antisense oligonucleotides abolished NE-induced PLD activation. The increase in PLD activity elicited by NE was also blocked by inhibitors of lipoxygenases (baicalein) and CYP4A (17-octadecynoic acid), but not of cyclooxygenase (indomethacin). AA and its metabolites (12(S)-, 15(S)-, and 20-HETEs) increased PLD activity. PLD activation by AA and HETEs was reduced by inhibitors of Ras farnesyltransferase (farnesyl protein transferase III and BMS-191563) and MEK (U0126 and PD98059). These data suggest that HETEs are the mediators of cPLA2-dependent PLD activation by NE in VSMC. In addition to cPLA2, PLD was also found to contribute to AA release for prostacyclin production via the phosphatidate phosphohydrolase/diacylglycerol lipase pathway. Finally, a catalytically inactive PLD2 (but not PLD1) mutant inhibited NE-induced PLD activity, and PLD2 was tyrosine-phosphorylated in response to NE by a MAPK-dependent pathway. We conclude that NE stimulates cPLA2-dependent PLD2 through lipoxygenase- and CYP4A-derived HETEs via the Ras/ERK pathway by a mechanism involving tyrosine phosphorylation of PLD2 in rabbit VSMC.

Original languageEnglish (US)
Pages (from-to)15704-15711
Number of pages8
JournalJournal of Biological Chemistry
Volume276
Issue number19
DOIs
StatePublished - May 11 2001

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Cytosolic Phospholipases A2
Phospholipase D
Phosphorylation
Hydroxyeicosatetraenoic Acids
Tyrosine
Norepinephrine
Chemical activation
Arachidonic Acid
Vascular Smooth Muscle
Smooth Muscle Myocytes
Muscle
Cytochrome P-450 Enzyme System
Calcium-Calmodulin-Dependent Protein Kinase Type 2
Lipoxygenase
Metabolites
calmidazolium
Phosphatidate Phosphatase
20-hydroxy-5,8,11,14-eicosatetraenoic acid
Rabbits
ras Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

@article{2f340301a076449694749b14ac33c293,
title = "Phospholipase D activation by norepinephrine is mediated by 12(S)-, 15(S)-, and 20-hydroxyeicosatetraenoic acids generated by stimulation of cytosolic phospholipase A2: Tyrosine phosphorylation of phospholipase D 2 in response to norepinephrine",
abstract = "Norepinephrine (NE) stimulates phospholipase D (PLD) through a Ras/MAPK pathway in rabbit vascular smooth muscle cells (VSMC). NE also activates calcium influx and calmodulin (CaM)-dependent protein kinase II-dependent cytosolic phospholipase A2 (cPLA2). Arachidonic acid (AA) released by cPLA2-catalyzed phospholipid hydrolysis is then metabolized into hydroxyeicosatetraenoic acids (HETEs) through lipoxygenase and cytochrome P450 4A (CYP4A) pathways. HETEs, in turn, have been shown to stimulate Ras translocation and to increase MAPK activity in VSMC. This study was conducted to determine the contribution of cPLA2-derived AA and its metabolites (HETEs) to the activation of PLD. NE-induced PLD activation was reduced by two structurally distinct CaM antagonists, W-7 and calmidazolium, and by CaM-dependent protein kinase II inhibition. Blockade of cPLA2 activity or protein depletion with selective cPLA2 antisense oligonucleotides abolished NE-induced PLD activation. The increase in PLD activity elicited by NE was also blocked by inhibitors of lipoxygenases (baicalein) and CYP4A (17-octadecynoic acid), but not of cyclooxygenase (indomethacin). AA and its metabolites (12(S)-, 15(S)-, and 20-HETEs) increased PLD activity. PLD activation by AA and HETEs was reduced by inhibitors of Ras farnesyltransferase (farnesyl protein transferase III and BMS-191563) and MEK (U0126 and PD98059). These data suggest that HETEs are the mediators of cPLA2-dependent PLD activation by NE in VSMC. In addition to cPLA2, PLD was also found to contribute to AA release for prostacyclin production via the phosphatidate phosphohydrolase/diacylglycerol lipase pathway. Finally, a catalytically inactive PLD2 (but not PLD1) mutant inhibited NE-induced PLD activity, and PLD2 was tyrosine-phosphorylated in response to NE by a MAPK-dependent pathway. We conclude that NE stimulates cPLA2-dependent PLD2 through lipoxygenase- and CYP4A-derived HETEs via the Ras/ERK pathway by a mechanism involving tyrosine phosphorylation of PLD2 in rabbit VSMC.",
author = "Parmentier, {Jean Hugues} and Muthalif, {Mubarack M.} and Saeed, {Abdelwahab E.} and Kafait Malik",
year = "2001",
month = "5",
day = "11",
doi = "10.1074/jbc.M011473200",
language = "English (US)",
volume = "276",
pages = "15704--15711",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "19",

}

TY - JOUR

T1 - Phospholipase D activation by norepinephrine is mediated by 12(S)-, 15(S)-, and 20-hydroxyeicosatetraenoic acids generated by stimulation of cytosolic phospholipase A2

T2 - Tyrosine phosphorylation of phospholipase D 2 in response to norepinephrine

AU - Parmentier, Jean Hugues

AU - Muthalif, Mubarack M.

AU - Saeed, Abdelwahab E.

AU - Malik, Kafait

PY - 2001/5/11

Y1 - 2001/5/11

N2 - Norepinephrine (NE) stimulates phospholipase D (PLD) through a Ras/MAPK pathway in rabbit vascular smooth muscle cells (VSMC). NE also activates calcium influx and calmodulin (CaM)-dependent protein kinase II-dependent cytosolic phospholipase A2 (cPLA2). Arachidonic acid (AA) released by cPLA2-catalyzed phospholipid hydrolysis is then metabolized into hydroxyeicosatetraenoic acids (HETEs) through lipoxygenase and cytochrome P450 4A (CYP4A) pathways. HETEs, in turn, have been shown to stimulate Ras translocation and to increase MAPK activity in VSMC. This study was conducted to determine the contribution of cPLA2-derived AA and its metabolites (HETEs) to the activation of PLD. NE-induced PLD activation was reduced by two structurally distinct CaM antagonists, W-7 and calmidazolium, and by CaM-dependent protein kinase II inhibition. Blockade of cPLA2 activity or protein depletion with selective cPLA2 antisense oligonucleotides abolished NE-induced PLD activation. The increase in PLD activity elicited by NE was also blocked by inhibitors of lipoxygenases (baicalein) and CYP4A (17-octadecynoic acid), but not of cyclooxygenase (indomethacin). AA and its metabolites (12(S)-, 15(S)-, and 20-HETEs) increased PLD activity. PLD activation by AA and HETEs was reduced by inhibitors of Ras farnesyltransferase (farnesyl protein transferase III and BMS-191563) and MEK (U0126 and PD98059). These data suggest that HETEs are the mediators of cPLA2-dependent PLD activation by NE in VSMC. In addition to cPLA2, PLD was also found to contribute to AA release for prostacyclin production via the phosphatidate phosphohydrolase/diacylglycerol lipase pathway. Finally, a catalytically inactive PLD2 (but not PLD1) mutant inhibited NE-induced PLD activity, and PLD2 was tyrosine-phosphorylated in response to NE by a MAPK-dependent pathway. We conclude that NE stimulates cPLA2-dependent PLD2 through lipoxygenase- and CYP4A-derived HETEs via the Ras/ERK pathway by a mechanism involving tyrosine phosphorylation of PLD2 in rabbit VSMC.

AB - Norepinephrine (NE) stimulates phospholipase D (PLD) through a Ras/MAPK pathway in rabbit vascular smooth muscle cells (VSMC). NE also activates calcium influx and calmodulin (CaM)-dependent protein kinase II-dependent cytosolic phospholipase A2 (cPLA2). Arachidonic acid (AA) released by cPLA2-catalyzed phospholipid hydrolysis is then metabolized into hydroxyeicosatetraenoic acids (HETEs) through lipoxygenase and cytochrome P450 4A (CYP4A) pathways. HETEs, in turn, have been shown to stimulate Ras translocation and to increase MAPK activity in VSMC. This study was conducted to determine the contribution of cPLA2-derived AA and its metabolites (HETEs) to the activation of PLD. NE-induced PLD activation was reduced by two structurally distinct CaM antagonists, W-7 and calmidazolium, and by CaM-dependent protein kinase II inhibition. Blockade of cPLA2 activity or protein depletion with selective cPLA2 antisense oligonucleotides abolished NE-induced PLD activation. The increase in PLD activity elicited by NE was also blocked by inhibitors of lipoxygenases (baicalein) and CYP4A (17-octadecynoic acid), but not of cyclooxygenase (indomethacin). AA and its metabolites (12(S)-, 15(S)-, and 20-HETEs) increased PLD activity. PLD activation by AA and HETEs was reduced by inhibitors of Ras farnesyltransferase (farnesyl protein transferase III and BMS-191563) and MEK (U0126 and PD98059). These data suggest that HETEs are the mediators of cPLA2-dependent PLD activation by NE in VSMC. In addition to cPLA2, PLD was also found to contribute to AA release for prostacyclin production via the phosphatidate phosphohydrolase/diacylglycerol lipase pathway. Finally, a catalytically inactive PLD2 (but not PLD1) mutant inhibited NE-induced PLD activity, and PLD2 was tyrosine-phosphorylated in response to NE by a MAPK-dependent pathway. We conclude that NE stimulates cPLA2-dependent PLD2 through lipoxygenase- and CYP4A-derived HETEs via the Ras/ERK pathway by a mechanism involving tyrosine phosphorylation of PLD2 in rabbit VSMC.

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U2 - 10.1074/jbc.M011473200

DO - 10.1074/jbc.M011473200

M3 - Article

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JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

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