Inhibition of the soluble epoxide hydrolase by tyrosine nitration

Eduardo Barbosa-Sicard, Timo Frömel, Benjamin Keserü, Ralf P. Brandes, Christophe Morisseau, Bruce D. Hammock, Thomas Braun, Marcus Krüger, Ingrid Fleming

Research output: Contribution to journalArticle

18 Citations (Scopus)

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 (Tyr383 and Tyr466) 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-morpholinosydnonimine (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 Tyr383 and Tyr466. 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.

Original languageEnglish (US)
Pages (from-to)28156-28163
Number of pages8
JournalJournal of Biological Chemistry
Volume284
Issue number41
DOIs
StatePublished - Oct 9 2009
Externally publishedYes

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Nitration
Epoxide Hydrolases
Tyrosine
Peroxynitrous Acid
Experimental Diabetes Mellitus
HEK293 Cells
Epoxy Compounds
Endothelial cells
Hydrolases
Streptozocin
Medical problems
Phenylalanine
Arachidonic Acid
Metabolism
Mass spectrometry
Blood Vessels
Mass Spectrometry
Proteins
Endothelial Cells
Hypertension

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Barbosa-Sicard, E., Frömel, T., Keserü, B., Brandes, R. P., Morisseau, C., Hammock, B. D., ... Fleming, I. (2009). Inhibition of the soluble epoxide hydrolase by tyrosine nitration. Journal of Biological Chemistry, 284(41), 28156-28163. https://doi.org/10.1074/jbc.M109.054759

Inhibition of the soluble epoxide hydrolase by tyrosine nitration. / Barbosa-Sicard, Eduardo; Frömel, Timo; Keserü, Benjamin; Brandes, Ralf P.; Morisseau, Christophe; Hammock, Bruce D.; Braun, Thomas; Krüger, Marcus; Fleming, Ingrid.

In: Journal of Biological Chemistry, Vol. 284, No. 41, 09.10.2009, p. 28156-28163.

Research output: Contribution to journalArticle

Barbosa-Sicard, E, Frömel, T, Keserü, B, Brandes, RP, Morisseau, C, Hammock, BD, Braun, T, Krüger, M & Fleming, I 2009, 'Inhibition of the soluble epoxide hydrolase by tyrosine nitration', Journal of Biological Chemistry, vol. 284, no. 41, pp. 28156-28163. https://doi.org/10.1074/jbc.M109.054759
Barbosa-Sicard E, Frömel T, Keserü B, Brandes RP, Morisseau C, Hammock BD et al. Inhibition of the soluble epoxide hydrolase by tyrosine nitration. Journal of Biological Chemistry. 2009 Oct 9;284(41):28156-28163. https://doi.org/10.1074/jbc.M109.054759
Barbosa-Sicard, Eduardo ; Frömel, Timo ; Keserü, Benjamin ; Brandes, Ralf P. ; Morisseau, Christophe ; Hammock, Bruce D. ; Braun, Thomas ; Krüger, Marcus ; Fleming, Ingrid. / Inhibition of the soluble epoxide hydrolase by tyrosine nitration. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 41. pp. 28156-28163.
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