NADP(+)-dependent dehydrogenase activity of carbonyl reductase on glutathionylhydroxynonanal as a new pathway for hydroxynonenal detoxification

Free Radic Biol Med. 2015 Jun:83:66-76. doi: 10.1016/j.freeradbiomed.2015.02.001. Epub 2015 Feb 11.

Abstract

An NADP(+)-dependent dehydrogenase activity on 3-glutathionyl-4-hydroxynonanal (GSHNE) was purified to electrophoretic homogeneity from a line of human astrocytoma cells (ADF). Proteomic analysis identified this enzymatic activity as associated with carbonyl reductase 1 (EC 1.1.1.184). The enzyme is highly efficient at catalyzing the oxidation of GSHNE (KM 33 µM, kcat 405 min(-1)), as it is practically inactive toward trans-4-hydroxy-2-nonenal (HNE) and other HNE-adducted thiol-containing amino acid derivatives. Combined mass spectrometry and nuclear magnetic resonance spectroscopy analysis of the reaction products revealed that carbonyl reductase oxidizes the hydroxyl group of GSHNE in its hemiacetal form, with the formation of the corresponding 3-glutathionylnonanoic-δ-lactone. The relevance of this new reaction catalyzed by carbonyl reductase 1 is discussed in terms of HNE detoxification and the recovery of reducing power.

Keywords: 3-Glutathionyl-4-hydroxynonanal; 4-Hydroxy-2-nonenal; Carbonyl reductase; Free radicals; Hydroxynonenal detoxification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alcohol Oxidoreductases / isolation & purification
  • Alcohol Oxidoreductases / metabolism*
  • Aldehyde Reductase / metabolism
  • Aldehydes / metabolism*
  • Astrocytoma / metabolism*
  • Astrocytoma / pathology
  • Glutathione / analogs & derivatives*
  • Glutathione / metabolism*
  • Humans
  • Inactivation, Metabolic*
  • Lactones / metabolism
  • Magnetic Resonance Spectroscopy
  • Mass Spectrometry
  • NADP / metabolism*
  • NADPH Dehydrogenase / metabolism*
  • Oxidation-Reduction
  • Proteomics
  • Substrate Specificity
  • Sulfhydryl Compounds / metabolism
  • Tumor Cells, Cultured

Substances

  • 3-glutathionyl-4-hydroxynonanal
  • Aldehydes
  • Lactones
  • Sulfhydryl Compounds
  • NADP
  • Alcohol Oxidoreductases
  • CBR1 protein, human
  • Aldehyde Reductase
  • NADPH Dehydrogenase
  • Glutathione
  • 4-hydroxy-2-nonenal