A 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 (E.C. 1.1.1.184). The enzyme is highly efficient at catalyzing the oxidation of GSHNE (KM33 µM,kcat.405 min-1), as it is practically inactive towards trans-4-hydroxy-2-nonenal (HNE) and other HNE-addicted 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-glutathionyl-nonanoic-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.

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

MOSCHINI, ROBERTA;PERONI, ELEONORA;CAPPIELLO, MARIO;MURA, UMBERTO;DEL CORSO, ANTONELLA
2015

Abstract

A 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 (E.C. 1.1.1.184). The enzyme is highly efficient at catalyzing the oxidation of GSHNE (KM33 µM,kcat.405 min-1), as it is practically inactive towards trans-4-hydroxy-2-nonenal (HNE) and other HNE-addicted 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-glutathionyl-nonanoic-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.
Moschini, Roberta; Peroni, Eleonora; Rotondo, Rossella; Renzone, G.; Melck, D.; Cappiello, Mario; Srebot, M.; Napolitano, E.; Motta, A.; Scaloni, A.; Mura, Umberto; DEL CORSO, Antonella
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11568/733869
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