GPR40 is a low-affinity epoxyeicosatrienoic acid receptor in vascular cells

Sang Kyu Park, Anja Herrnreiter, Sandra L. Pfister, Kathryn M. Gauthier, Benjamin A. Falck, J R Falck, William B. Campbell

Research output: Contribution to journalArticle

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Abstract

Endothelium-derived epoxyeicosatrienoic acids (EETs) have numerous vascular activities mediated by G protein–coupled receptors. Long-chain free fatty acids and EETs activate GPR40, prompting us to investigate the role of GPR40 in some vascular EET activities. 14,15-EET, 11,12-EET, arachidonic acid, and the GPR40 agonist GW9508 increase intracellular calcium concentrations in human GPR40 – overexpressing HEK293 cells (EC50 0.58 0.08 M, 0.91 0.08 M, 3.9 0.06 M, and 19 0.37 nM, respectively). EETs with cis- and trans-epoxides had similar activities, whereas substitution of a thiirane sulfur for the epoxide oxygen decreased the activities. 8,9-EET, 5,6-EET, and the epoxide hydrolysis products 11,12- and 14,15-dihydroxyeico-satrienoic acids were less active than 11,12-EET. The GPR40 antagonist GW1100 and siRNA-mediated GPR40 silencing blocked the EET- and GW9508-induced calcium increases. EETs are weak GPR120 agonists. GPR40 expression was detected in human and bovine endothelial cells (ECs), smooth muscle cells, and arteries. 11,12-EET concentration-dependently relaxed preconstricted coronary arteries; however, these relaxations were not altered by GW1100. In human ECs, 11,12-EET increased MAP kinase (MAPK)-mediated ERK phosphorylation, phosphorylation and levels of connexin-43 (Cx43), and expression of cyclooxygenase-2 (COX-2), all of which were inhibited by GW1100 and the MAPK inhibitor U0126. Moreover, siRNA-mediated GPR40 silencing decreased 11,12-EET–induced ERK phosphorylation. These results indicated that GPR40 is a low-affinity EET receptor in vascular cells and arteries. We conclude that epoxidation of arachidonic acid to EETs enhances GPR40 agonist activity and that 11,12-EET stimulation of GPR40 increases Cx43 and COX-2 expression in ECs via ERK phosphorylation.

Original languageEnglish (US)
Pages (from-to)10675-10691
Number of pages17
JournalJournal of Biological Chemistry
Volume293
Issue number27
DOIs
StatePublished - Jan 1 2018

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Phosphorylation
Blood Vessels
Epoxy Compounds
Endothelial cells
Acids
Connexin 43
Endothelial Cells
Cyclooxygenase 2
Arachidonic Acid
Small Interfering RNA
Phosphotransferases
Arteries
Calcium
Epoxidation
HEK293 Cells
Sulfur
Nonesterified Fatty Acids
Smooth Muscle Myocytes
Endothelium
Muscle

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Park, S. K., Herrnreiter, A., Pfister, S. L., Gauthier, K. M., Falck, B. A., Falck, J. R., & Campbell, W. B. (2018). GPR40 is a low-affinity epoxyeicosatrienoic acid receptor in vascular cells. Journal of Biological Chemistry, 293(27), 10675-10691. https://doi.org/10.1074/jbc.RA117.001297

GPR40 is a low-affinity epoxyeicosatrienoic acid receptor in vascular cells. / Park, Sang Kyu; Herrnreiter, Anja; Pfister, Sandra L.; Gauthier, Kathryn M.; Falck, Benjamin A.; Falck, J R; Campbell, William B.

In: Journal of Biological Chemistry, Vol. 293, No. 27, 01.01.2018, p. 10675-10691.

Research output: Contribution to journalArticle

Park, SK, Herrnreiter, A, Pfister, SL, Gauthier, KM, Falck, BA, Falck, JR & Campbell, WB 2018, 'GPR40 is a low-affinity epoxyeicosatrienoic acid receptor in vascular cells', Journal of Biological Chemistry, vol. 293, no. 27, pp. 10675-10691. https://doi.org/10.1074/jbc.RA117.001297
Park, Sang Kyu ; Herrnreiter, Anja ; Pfister, Sandra L. ; Gauthier, Kathryn M. ; Falck, Benjamin A. ; Falck, J R ; Campbell, William B. / GPR40 is a low-affinity epoxyeicosatrienoic acid receptor in vascular cells. In: Journal of Biological Chemistry. 2018 ; Vol. 293, No. 27. pp. 10675-10691.
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abstract = "Endothelium-derived epoxyeicosatrienoic acids (EETs) have numerous vascular activities mediated by G protein–coupled receptors. Long-chain free fatty acids and EETs activate GPR40, prompting us to investigate the role of GPR40 in some vascular EET activities. 14,15-EET, 11,12-EET, arachidonic acid, and the GPR40 agonist GW9508 increase intracellular calcium concentrations in human GPR40 – overexpressing HEK293 cells (EC50 0.58 0.08 M, 0.91 0.08 M, 3.9 0.06 M, and 19 0.37 nM, respectively). EETs with cis- and trans-epoxides had similar activities, whereas substitution of a thiirane sulfur for the epoxide oxygen decreased the activities. 8,9-EET, 5,6-EET, and the epoxide hydrolysis products 11,12- and 14,15-dihydroxyeico-satrienoic acids were less active than 11,12-EET. The GPR40 antagonist GW1100 and siRNA-mediated GPR40 silencing blocked the EET- and GW9508-induced calcium increases. EETs are weak GPR120 agonists. GPR40 expression was detected in human and bovine endothelial cells (ECs), smooth muscle cells, and arteries. 11,12-EET concentration-dependently relaxed preconstricted coronary arteries; however, these relaxations were not altered by GW1100. In human ECs, 11,12-EET increased MAP kinase (MAPK)-mediated ERK phosphorylation, phosphorylation and levels of connexin-43 (Cx43), and expression of cyclooxygenase-2 (COX-2), all of which were inhibited by GW1100 and the MAPK inhibitor U0126. Moreover, siRNA-mediated GPR40 silencing decreased 11,12-EET–induced ERK phosphorylation. These results indicated that GPR40 is a low-affinity EET receptor in vascular cells and arteries. We conclude that epoxidation of arachidonic acid to EETs enhances GPR40 agonist activity and that 11,12-EET stimulation of GPR40 increases Cx43 and COX-2 expression in ECs via ERK phosphorylation.",
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N2 - Endothelium-derived epoxyeicosatrienoic acids (EETs) have numerous vascular activities mediated by G protein–coupled receptors. Long-chain free fatty acids and EETs activate GPR40, prompting us to investigate the role of GPR40 in some vascular EET activities. 14,15-EET, 11,12-EET, arachidonic acid, and the GPR40 agonist GW9508 increase intracellular calcium concentrations in human GPR40 – overexpressing HEK293 cells (EC50 0.58 0.08 M, 0.91 0.08 M, 3.9 0.06 M, and 19 0.37 nM, respectively). EETs with cis- and trans-epoxides had similar activities, whereas substitution of a thiirane sulfur for the epoxide oxygen decreased the activities. 8,9-EET, 5,6-EET, and the epoxide hydrolysis products 11,12- and 14,15-dihydroxyeico-satrienoic acids were less active than 11,12-EET. The GPR40 antagonist GW1100 and siRNA-mediated GPR40 silencing blocked the EET- and GW9508-induced calcium increases. EETs are weak GPR120 agonists. GPR40 expression was detected in human and bovine endothelial cells (ECs), smooth muscle cells, and arteries. 11,12-EET concentration-dependently relaxed preconstricted coronary arteries; however, these relaxations were not altered by GW1100. In human ECs, 11,12-EET increased MAP kinase (MAPK)-mediated ERK phosphorylation, phosphorylation and levels of connexin-43 (Cx43), and expression of cyclooxygenase-2 (COX-2), all of which were inhibited by GW1100 and the MAPK inhibitor U0126. Moreover, siRNA-mediated GPR40 silencing decreased 11,12-EET–induced ERK phosphorylation. These results indicated that GPR40 is a low-affinity EET receptor in vascular cells and arteries. We conclude that epoxidation of arachidonic acid to EETs enhances GPR40 agonist activity and that 11,12-EET stimulation of GPR40 increases Cx43 and COX-2 expression in ECs via ERK phosphorylation.

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