Journal article
Human carbonyl reductase catalyzes reduction of 4-oxonon-2-enal
Biochemistry (Easton), Vol.43(41), pp.13106-13114
10/19/2004
DOI: 10.1021/bi049136q
PMID: 15476404
Abstract
4-Oxonon-2-enal (4ONE) was demonstrated to be a product of lipid peroxidation, and previous studies found that it was highly reactive toward DNA and protein. The present study sought to determine whether carbonyl reductase (CR) catalyzes reduction of 4ONE, representing a potential pathway for metabolism of the lipid peroxidation product. Recombinant CR was cloned from a human liver cDNA library, expressed in Escherichia coli, and purified by metal chelate chromatography. Both 4ONE and its glutathione conjugate were found to be substrates for CR, and kinetic parameters were calculated. TLC analysis of reaction products revealed the presence of three compounds, two of which were identified as 4-hydroxynon-2-enal (4HNE) and 1-hydroxynon-2-en-4-one (1HNO). GC/MS analysis confirmed 4HNE and 1HNO and identified the unknown reaction product as 4-oxononanal (4ONA). Analysis of oxime derivatives of the reaction products via LC/MS confirmed the unknown as 4ONA. The time course for CR-mediated, NADPH-dependent 4ONE reduction and appearance of 4HNE and 1HNO was determined using HPLC, demonstrating 4HNE to be a major product and 1HNO and 4ONA to be minor products. Simulated structures of 4ONE in the active site of CR/NADPH calculated via docking experiments predict the ketone positioned as primary hydride acceptor. Results of the present study demonstrate that 4ONE is a substrate for CR/NADPH and the enzyme may represent a pathway for biotransformation of the lipid. Furthermore, these findings reveal that CR catalyzes hydride transfer selectively to the ketone but also to the aldehyde and C=C of 4ONE, resulting in 4HNE, 1HNO, and 4ONA, respectively.
Details
- Title: Subtitle
- Human carbonyl reductase catalyzes reduction of 4-oxonon-2-enal
- Creators
- Jonathan A Doorn - Department of Pharmaceutical Sciences, School of Pharmacy, The University of Colorado Health Sciences Center, Denver, Colorado 80262, USAEdmund MaserAndreas BlumDavid J ClaffeyDennis R Petersen
- Resource Type
- Journal article
- Publication Details
- Biochemistry (Easton), Vol.43(41), pp.13106-13114
- Publisher
- United States
- DOI
- 10.1021/bi049136q
- PMID
- 15476404
- ISSN
- 0006-2960
- eISSN
- 1520-4995
- Grant note
- R01AA09300 / NIAAA NIH HHS F32 ES11937 / NIEHS NIH HHS R01ES09410 / NIEHS NIH HHS
- Language
- English
- Date published
- 10/19/2004
- Academic Unit
- Iowa Neuroscience Institute; Pharmaceutical Sciences and Experimental Therapeutics; Medicinal and Natural Products Chemistry
- Record Identifier
- 9984070286802771
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