title: Inhibition of the soluble epoxide hydrolase by tyrosine nitration.
abstract: Inhibition of the soluble epoxide hydrolase (sEH) has beneficial effects on vascular inflammation and hypertension indicating that the enzyme may be a promising target for drug development. As the enzymatic core of the hydrolase domain of the human sEH contains two tyrosine residues (Tyr(383) and Tyr(466)) that are theoretically crucial for enzymatic activity, we addressed the hypothesis that the activity of the sEH may be affected by nitrosative stress. Epoxide hydrolase activity was detected in human and murine endothelial cells as well in HEK293 cells and could be inhibited by either authentic peroxynitrite (ONOO(-)) or the ONOO(-) generator 3-morpholino-sydnonimine (SIN-1). Protection of the enzymatic core with 1-adamantyl-3-cyclohexylurea in vitro decreased sensitivity to SIN-1. Both ONOO(-) and SIN-1 elicited the tyrosine nitration of the sEH protein and mass spectrometry analysis of tryptic fragments revealed nitration on several tyrosine residues including Tyr(383) and Tyr(466). Mutation of the latter residues to phenylalanine was sufficient to abrogate epoxide hydrolase activity. In vivo, streptozotocin-induced diabetes resulted in the tyrosine nitration of the sEH in murine lungs and a significant decrease in its activity. Taken together, these data indicate that the activity of the sEH can be regulated by the tyrosine nitration of the protein. Moreover, nitrosative stress would be expected to potentiate the physiological actions of arachidonic acid epoxides by preventing their metabolism to the corresponding diols.
authors.affiliation: Institute for Vascular Signalling, Johann Wolfgang Goethe University, D-60590 Frankfurt am Main, Germany.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Barbosa-Sicard
authors.affiliation: Institute for Vascular Signalling, Johann Wolfgang Goethe University, D-60590 Frankfurt am Main, Germany.
authors.forename: Timo
authors.initials: T
authors.lastname: Frömel
authors.affiliation: Institute for Vascular Signalling, Johann Wolfgang Goethe University, D-60590 Frankfurt am Main, Germany.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Keserü
authors.affiliation: Institut für Kardiovaskuläre Physiologie, Johann Wolfgang Goethe University, D-60590 Frankfurt am Main, Germany.
authors.forename: Ralf P
authors.initials: RP
authors.lastname: Brandes
authors.affiliation: Department of Entomology, University of California, Davis, California 95616.
authors.forename: Christophe
authors.initials: C
authors.lastname: Morisseau
authors.affiliation: Department of Entomology, University of California, Davis, California 95616.
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Hammock
authors.affiliation: Department of Cardiac Development and Remodeling, Max-Planck-Institut for Heart and Lung Research, D-61231 Bad Nauheim, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Braun
authors.affiliation: Department of Cardiac Development and Remodeling, Max-Planck-Institut for Heart and Lung Research, D-61231 Bad Nauheim, Germany.
authors.forename: Marcus
authors.initials: M
authors.lastname: Krüger
authors.affiliation: Institute for Vascular Signalling, Johann Wolfgang Goethe University, D-60590 Frankfurt am Main, Germany. Electronic address: fleming@em.uni-frankfurt.de.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Fleming
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000201:enzymology; D004791:Enzyme Inhibitors / Q000378:metabolism; D004851:Epoxide Hydrolases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008820:Mice, Obese; D008981:Molsidomine / Q000031:analogs & derivatives / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D018384:Oxidative Stress; D030421:Peroxynitrous Acid / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D030421:Peroxynitrous Acid; D014443:Tyrosine; C002385:linsidomine; D008981:Molsidomine; D004851:Epoxide Hydrolases
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