Nrf2-mediated induction of cytoprotective enzymes by 15-deoxy-Delta12,14-prostaglandin J2 is attenuated by alkenal/one oxidoreductase

J Biol Chem. 2006 Sep 8;281(36):26245-52. doi: 10.1074/jbc.M604620200. Epub 2006 Jul 20.

Abstract

NADPH-dependent alkenal/one oxidoreductase (Aor) was discovered to be highly inducible in rat liver following treatment with the cancer chemopreventive agent 3H-1, 2-dithiole-3-thione. Aor was further characterized as an Nrf2-regulated antioxidative enzyme that reduces carbon-carbon double bonds in a variety of alpha, beta-unsaturated aldehydes and ketones. 15-Deoxy-Delta12,14-prostaglandin J2 (15d-PGJ2) is a reactive membrane lipid metabolite that activates multiple pathways, including Nrf2-mediated induction of cytoprotective enzymes. Physiologically, it is postulated that 15d-PGJ2 alkylates key regulatory proteins via the electrophilic carbon centers found in two alpha, beta-unsaturated ketone moieties. This current study addresses the metabolism of 15d-PGJ2 by rat Aor (rAor) and subsequent deactivation of the Nrf2 signaling pathway by both rat and human AOR. We demonstrate that induction of NADPH-dependent quinone oxidoreductase activity by 15d-PGJ2 is markedly attenuated in mouse embryonic fibroblasts that overexpress rAor. Luciferase reporter assay and quantitative real-time PCR confirmed these findings. Concentrations required for doubling the NADPH-dependent quinone oxidoreductase response are increased from 1.8 microm in wild-type to >10 microm in rat Aor transgenic fibroblasts. 15d-PGJ2 is metabolized by recombinant rAor with a Km of 9.6 microm and k(cat) of 18.5 min(-1). The major product is 12,13-dihydro-15-deoxy-Delta12,14-prostaglandin J2 (dihydro-15d-PGJ2). The reduction of C=C by Aor yielding dihydro-15d-PGJ2 abolishes the inducibility in an antioxidant response element-driven luciferase assay. Collectively, these results demonstrate that 15d-PGJ2 can be catabolized by Aor, thereby attenuating subsequent Nrf2 signaling and possibly inflammatory and apoptotic processes also influenced by 15d-PGJ2.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Alcohol Oxidoreductases / genetics
  • Alcohol Oxidoreductases / metabolism*
  • Animals
  • Cells, Cultured
  • Enzyme Induction*
  • Fibroblasts / cytology
  • Fibroblasts / physiology
  • Humans
  • Immunologic Factors / metabolism*
  • Mice
  • Mice, Transgenic
  • Molecular Structure
  • NAD(P)H Dehydrogenase (Quinone) / genetics
  • NAD(P)H Dehydrogenase (Quinone) / metabolism*
  • NF-E2-Related Factor 2 / genetics
  • NF-E2-Related Factor 2 / metabolism*
  • Oxidation-Reduction
  • Prostaglandin D2 / analogs & derivatives*
  • Prostaglandin D2 / chemistry
  • Prostaglandin D2 / genetics
  • Prostaglandin D2 / metabolism
  • Rats
  • Signal Transduction / physiology

Substances

  • 15-deoxy-delta(12,14)-prostaglandin J2
  • Immunologic Factors
  • NF-E2-Related Factor 2
  • NFE2L2 protein, human
  • Alcohol Oxidoreductases
  • leukotriene B4 12-hydroxydehydrogenase
  • NAD(P)H Dehydrogenase (Quinone)
  • Prostaglandin D2