title: Functional screening for G protein-coupled receptor targets of 14,15-epoxyeicosatrienoic acid.
abstract: Epoxyeicosatrienoic acids (EETs) are potent vasodilators that play important roles in cardiovascular physiology and disease, yet the molecular mechanisms underlying the biological actions of EETs are not fully understood. Multiple lines of evidence suggest that the actions of EETs are in part mediated via G protein-coupled receptor (GPCR) signaling, but the identity of such a receptor has remained elusive. We sought to identify 14,15-EET-responsive GPCRs. A set of 105 clones were expressed in Xenopus oocyte and screened for their ability to activate cAMP-dependent chloride current. Several receptors responded to micromolar concentrations of 14,15-EET, with the top five being prostaglandin receptor subtypes (PTGER2, PTGER4, PTGFR, PTGDR, PTGER3IV). Overall, our results indicate that multiple low-affinity 14,15-EET GPCRs are capable of increasing cAMP levels following 14,15-EET stimulation, highlighting the potential for cross-talk between prostanoid and other ecosanoid GPCRs. Our data also indicate that none of the 105 GPCRs screened met our criteria for a high-affinity receptor for 14,15-EET.
authors.affiliation: The Knight Cardiovascular Institute, Oregon Health & Science University, Portland, OR, United States.
authors.forename: Xuehong
authors.initials: X
authors.lastname: Liu
authors.affiliation: The Knight Cardiovascular Institute, Oregon Health & Science University, Portland, OR, United States.
authors.forename: Zu-Yuan
authors.initials: ZY
authors.lastname: Qian
authors.affiliation: Departments of Physiology & Pharmacology, Oregon Health & Science University, Portland, OR, United States.
authors.forename: Fuchun
authors.initials: F
authors.lastname: Xie
authors.affiliation: The Knight Cardiovascular Institute, Oregon Health & Science University, Portland, OR, United States.
authors.forename: Wei
authors.initials: W
authors.lastname: Fan
authors.affiliation: The Knight Cardiovascular Institute, Oregon Health & Science University, Portland, OR, United States.
authors.forename: Jonathan W
authors.initials: JW
authors.lastname: Nelson
authors.affiliation: Departments of Physiology & Pharmacology, Oregon Health & Science University, Portland, OR, United States; The Knight Cardiovascular Institute, Oregon Health & Science University, Portland, OR, United States.
authors.forename: Xiangshu
authors.initials: X
authors.lastname: Xiao
authors.affiliation: The Knight Cardiovascular Institute, Oregon Health & Science University, Portland, OR, United States.
authors.forename: Sanjiv
authors.initials: S
authors.lastname: Kaul
authors.affiliation: Departments of Pediatrics, Oregon Health & Science University, Portland, OR, United States; The Knight Cardiovascular Institute, Oregon Health & Science University, Portland, OR, United States. Electronic address: barnesan@ohsu.edu.
authors.forename: Anthony P
authors.initials: AP
authors.lastname: Barnes
authors.affiliation: Departments of Anesthesiology & Perioperative Medicine, Oregon Health & Science University, Portland, OR, United States; Departments of Physiology & Pharmacology, Oregon Health & Science University, Portland, OR, United States; The Knight Cardiovascular Institute, Oregon Health & Science University, Portland, OR, United States. Electronic address: alkayedn@ohsu.edu.
authors.forename: Nabil J
authors.initials: NJ
authors.lastname: Alkayed
mesh_terms: D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid / Q000494:pharmacology; D015126:8,11,14-Eicosatrienoic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D000242:Cyclic AMP / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism; D004353:Drug Evaluation, Preclinical; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D008638:Mesenteric Arteries / Q000187:drug effects / Q000502:physiology; D051379:Mice; D009865:Oocytes / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D011982:Receptors, Prostaglandin / Q000378:metabolism; D058307:Receptors, Prostaglandin E, EP2 Subtype / Q000378:metabolism; D058309:Receptors, Prostaglandin E, EP4 Subtype / Q000378:metabolism; D014661:Vasoconstriction / Q000187:drug effects; D014981:Xenopus; D000071557:beta-Arrestins / Q000378:metabolism
chemical_list: C545644:PTGER2 protein, human; C545650:PTGER4 protein, human; D043562:Receptors, G-Protein-Coupled; D011982:Receptors, Prostaglandin; D058307:Receptors, Prostaglandin E, EP2 Subtype; D058309:Receptors, Prostaglandin E, EP4 Subtype; D000071557:beta-Arrestins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; C046782:14,15-epoxy-5,8,11-eicosatrienoic acid; D000242:Cyclic AMP; D048049:Extracellular Signal-Regulated MAP Kinases; D015126:8,11,14-Eicosatrienoic Acid
keywords: CFTR; EET; Epoxyeicosatrienoic acid; GPCR; Prostaglandin E(2); Vasodilation