Aldose Reductase Catalyzes Reduction of the Lipid Peroxidation Product 4-Oxonon-2-enal

Jonathan A. Doorn, Satish Srivastava, Dennis R. Petersen

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Recent studies found that 4-oxonon-2-enal (4ONE) is a highly reactive product of lipid peroxidation that can modify peptides and protein sulfhydryls. Because aldose reductase (AR) was shown earlier to catalyze reduction of an α,β-unsaturated lipid aldehyde, 4-hydroxynon-2-enal (4HNE), it was systematically investigated if this enzyme could represent a pathway for 4ONE metabolism as well. 4ONE, the glutathione (GSH) conjugate of 4ONE (GS-4ONE), and 1-hydroxynonen-4-one (1HNO), the predicted initial metabolite of 4ONE reduction, were incubated with AR and NADPH, and kinetic constants were measured. The initial product of AR-mediated 4ONE reduction was identified as 1HNO, which could be further reduced to DHN, catalyzed by AR. This result indicates that the order of 4ONE carbonyl reduction is aldehyde and then ketone. 1HNO was found to be an electrophile toward GSH with reactivity ∼55-fold less than 4ONE but ∼2-fold higher than that of 4HNE. The enzyme had activity toward GS-4ONE, exhibiting a ∼4-fold higher κ cat/KM for GS-4ONE as compared to 4ONE. In the presence ofDepartment of Drug Metabolism NADPH, 4ONE did not inactivate AR, whereas in the absence of the cofactor, ∼60% of the enzyme activity was lost. The orientation of 4ONE in the AR active site was predicted using molecular modeling to explain the reactivity of 4ONE toward the enzyme. These simulations revealed that concurrent with NADPH binding to AR, Cys 298 is oriented such that the thiol group will not interact with 4ONE. Results of the present study are the first to demonstrate that AR may represent a pathway for metabolism of 4ONE and GS-4ONE.

Original languageEnglish (US)
Pages (from-to)1418-1423
Number of pages6
JournalChemical Research in Toxicology
Volume16
Issue number11
DOIs
StatePublished - Nov 2003

Fingerprint

Aldehyde Reductase
Lipid Peroxidation
Lipids
NADP
Metabolism
Enzyme activity
4-oxo-2-nonenal
Aldehydes
Enzymes
Molecular modeling
Coenzymes
Metabolites
Ketones
Sulfhydryl Compounds
Glutathione

ASJC Scopus subject areas

  • Drug Discovery
  • Organic Chemistry
  • Chemistry(all)
  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

Aldose Reductase Catalyzes Reduction of the Lipid Peroxidation Product 4-Oxonon-2-enal. / Doorn, Jonathan A.; Srivastava, Satish; Petersen, Dennis R.

In: Chemical Research in Toxicology, Vol. 16, No. 11, 11.2003, p. 1418-1423.

Research output: Contribution to journalArticle

Doorn, Jonathan A. ; Srivastava, Satish ; Petersen, Dennis R. / Aldose Reductase Catalyzes Reduction of the Lipid Peroxidation Product 4-Oxonon-2-enal. In: Chemical Research in Toxicology. 2003 ; Vol. 16, No. 11. pp. 1418-1423.
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abstract = "Recent studies found that 4-oxonon-2-enal (4ONE) is a highly reactive product of lipid peroxidation that can modify peptides and protein sulfhydryls. Because aldose reductase (AR) was shown earlier to catalyze reduction of an α,β-unsaturated lipid aldehyde, 4-hydroxynon-2-enal (4HNE), it was systematically investigated if this enzyme could represent a pathway for 4ONE metabolism as well. 4ONE, the glutathione (GSH) conjugate of 4ONE (GS-4ONE), and 1-hydroxynonen-4-one (1HNO), the predicted initial metabolite of 4ONE reduction, were incubated with AR and NADPH, and kinetic constants were measured. The initial product of AR-mediated 4ONE reduction was identified as 1HNO, which could be further reduced to DHN, catalyzed by AR. This result indicates that the order of 4ONE carbonyl reduction is aldehyde and then ketone. 1HNO was found to be an electrophile toward GSH with reactivity ∼55-fold less than 4ONE but ∼2-fold higher than that of 4HNE. The enzyme had activity toward GS-4ONE, exhibiting a ∼4-fold higher κ cat/KM for GS-4ONE as compared to 4ONE. In the presence ofDepartment of Drug Metabolism NADPH, 4ONE did not inactivate AR, whereas in the absence of the cofactor, ∼60{\%} of the enzyme activity was lost. The orientation of 4ONE in the AR active site was predicted using molecular modeling to explain the reactivity of 4ONE toward the enzyme. These simulations revealed that concurrent with NADPH binding to AR, Cys 298 is oriented such that the thiol group will not interact with 4ONE. Results of the present study are the first to demonstrate that AR may represent a pathway for metabolism of 4ONE and GS-4ONE.",
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AB - Recent studies found that 4-oxonon-2-enal (4ONE) is a highly reactive product of lipid peroxidation that can modify peptides and protein sulfhydryls. Because aldose reductase (AR) was shown earlier to catalyze reduction of an α,β-unsaturated lipid aldehyde, 4-hydroxynon-2-enal (4HNE), it was systematically investigated if this enzyme could represent a pathway for 4ONE metabolism as well. 4ONE, the glutathione (GSH) conjugate of 4ONE (GS-4ONE), and 1-hydroxynonen-4-one (1HNO), the predicted initial metabolite of 4ONE reduction, were incubated with AR and NADPH, and kinetic constants were measured. The initial product of AR-mediated 4ONE reduction was identified as 1HNO, which could be further reduced to DHN, catalyzed by AR. This result indicates that the order of 4ONE carbonyl reduction is aldehyde and then ketone. 1HNO was found to be an electrophile toward GSH with reactivity ∼55-fold less than 4ONE but ∼2-fold higher than that of 4HNE. The enzyme had activity toward GS-4ONE, exhibiting a ∼4-fold higher κ cat/KM for GS-4ONE as compared to 4ONE. In the presence ofDepartment of Drug Metabolism NADPH, 4ONE did not inactivate AR, whereas in the absence of the cofactor, ∼60% of the enzyme activity was lost. The orientation of 4ONE in the AR active site was predicted using molecular modeling to explain the reactivity of 4ONE toward the enzyme. These simulations revealed that concurrent with NADPH binding to AR, Cys 298 is oriented such that the thiol group will not interact with 4ONE. Results of the present study are the first to demonstrate that AR may represent a pathway for metabolism of 4ONE and GS-4ONE.

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