Inhibition of the soluble epoxide hydrolase by tyrosine nitration.
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.
Barbosa-Sicard
Eduardo
E
Institute for Vascular Signalling, Johann Wolfgang Goethe University, D-60590 Frankfurt am Main, Germany.
Frömel
Timo
T
Institute for Vascular Signalling, Johann Wolfgang Goethe University, D-60590 Frankfurt am Main, Germany.
Keserü
Benjamin
B
Institute for Vascular Signalling, Johann Wolfgang Goethe University, D-60590 Frankfurt am Main, Germany.
Brandes
Ralf P
RP
Institut für Kardiovaskuläre Physiologie, Johann Wolfgang Goethe University, D-60590 Frankfurt am Main, Germany.
Morisseau
Christophe
C
Department of Entomology, University of California, Davis, California 95616.
Hammock
Bruce D
BD
Department of Entomology, University of California, Davis, California 95616.
Braun
Thomas
T
Department of Cardiac Development and Remodeling, Max-Planck-Institut for Heart and Lung Research, D-61231 Bad Nauheim, Germany.
Krüger
Marcus
M
Department of Cardiac Development and Remodeling, Max-Planck-Institut for Heart and Lung Research, D-61231 Bad Nauheim, Germany.
Fleming
Ingrid
I
Institute for Vascular Signalling, Johann Wolfgang Goethe University, D-60590 Frankfurt am Main, Germany. Electronic address: fleming@em.uni-frankfurt.de.
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*
D004791:Enzyme Inhibitors; D030421:Peroxynitrous Acid; D014443:Tyrosine; C002385:linsidomine; D008981:Molsidomine; D004851:Epoxide Hydrolases
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