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Cited by 294Antioxidant Activity and Oxidative StressRedox biology and oxidative stressParaoxonase enzyme and polymorphisms

Quantification of malondialdehyde and 4-hydroxynonenal adducts to lysine residues in native and oxidized human low-density lipoprotein

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REQUENA, Jesús R., FU, Min Xin, AHMED, Mahtab U., JENKINS, Alicia J., LYONS, Timothy J., BAYNES, John W., THORPE, Suzanne R.·1997·Biochemical Journal
DOI 10.1042/bj3220317PMC PMC1218194
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Abstract

Malondialdehyde (MDA) and 4-hydroxynonenal (HNE) are major end-products of oxidation of polyunsaturated fatty acids, and are frequently measured as indicators of lipid peroxidation and oxidative stress in vivo. MDA forms Schiff-base adducts with lysine residues and cross-links proteins in vitro; HNE also reacts with lysines, primarily via a Michael addition reaction. We have developed methods using NaBH4 reduction to stabilize these adducts to conditions used for acid hydrolysis of protein, and have prepared reduced forms of lysine-MDA [3-(Nε-lysino)propan-1-ol (LM)], the lysine-MDA-lysine iminopropene cross-link [1,3-di(Nε-lysino)propane (LML)] and lysine-HNE [3-(Nε-lysino)-4-hydroxynonan-1-ol (LHNE)]. Gas chromatography/MS assays have been developed for quantification of the reduced compounds in protein. RNase incubated with MDA or HNE was used as a model for quantification of the adducts by gas chromatography/MS. There was excellent agreement between measurement of MDA bound to RNase as LM and LML, and as thiobarbituric acid-MDA adducts measured by HPLC; these adducts accounted for 70Ő80% of total lysine loss during the reaction with MDA. LM and LML (0.002Ő0.12mmol/mol of lysine) were also found in freshly isolated low-density lipoprotein (LDL) from healthy subjects. LHNE was measured in RNase treated with HNE, but was not detectable in native LDL. LM, LML and LHNE increased in concert with the formation of conjugated dienes during the copper-catalysed oxidation of LDL, but accounted for modification of < 1% of lysine residues in oxidized LDL. These results are the first report of direct chemical measurement of MDA and HNE adducts to lysine residues in LDL. LM, LML and LHNE should be useful as biomarkers of lipid peroxidative modification of protein and of oxidative stress in vitro and in vivo.

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  • Antibodies against MDA-modified proteins are commercially available and highly selective MS methods for their detection have been established.

    “To our knowledge, MDA modified proteins have not been used as biomarkers in recent clinical investigations although antibodies are commercially available and highly selective MS methods have been established, previously [57].”

    From Protein oxidation - Formation mechanisms, detection and relevance as biomarkers in human diseases (2021) · cites this paper as [57]

    The Introduction (page not provided) says antibodies are available for measuring MDA epitopes and describes a sensitive, specific GC/MS assay for MDA–lysine adducts in proteins. It does not say the antibodies are commercially available. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • 4-Hydroxy-2-nonenal (4-HNE) forms stable Michael adducts with cysteine and histidine; nucleophilic attack of lysine forms unstable Michael adducts that require reduction prior to analysis.

    “This positive partial charge is attacked by nucleophiles such as cysteine and histidine and stable Michael adducts are formed (Fig. 4B) [50]. Nucleophilic attack of lysine forms unstable Michael adducts, which require reduction, e.g., by sodium borohydride prior to analysis [57].”

    From Protein oxidation - Formation mechanisms, detection and relevance as biomarkers in human diseases (2021) · cites this paper as [57]

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2 of the 35 references couldn’t be matched to a known paper.

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  1. Fruebis, J., Parthasarathy, S. and Steinberg, D. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 10588-10592
  2. Sell, D. R. and Monnier, V. M. (1989) in Physiology of Aging (Masero, E. J., ed.), pp. 235-305, Oxford University Press, New York

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Authors
REQUENA, J. R., FU, M. X., AHMED, M. U., JENKINS, A. J., LYONS, T. J., BAYNES, J. W., & THORPE, S. R.
Affiliations
  • University of South Carolina (US)
  • Medical University of South Carolina (US)
ORCID
Venue
Biochemical Journal
Publisher
Portland Press Ltd.
Published
1997-02-15
Volume
322
Issue
1
Pages
317-325
ISSN
  • 0264-6021 (ISSN-L)
  • 1470-8728
DOI
10.1042/bj3220317
PMC
PMC1218194
OpenAlex
W1843398852
Cited by
293
References
35
Citations by year
2026: 4 · 2025: 3 · 2024: 3 · 2023: 13 · 2022: 7 · 2021: 6 · 2020: 7 · 2019: 2 · 2018: 5 · 2017: 9 · 2016: 6 · 2015: 5 · 2014: 7 · 2013: 11 · 2012: 14
FWCI
10.07
Type
journal article
Language
en
Open access
Locations
  • Biochemical Journal (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Scholar Commons (University of South Carolina) (submittedVersion)
  • Research Portal (Queen's University Belfast) (submittedVersion)
Funders
  • National Institutes of Health (5r01ag011472-04)
  • National Institute on Aging (AG-11472)
  • National Eye Institute (5R29 EY10697, R29EY010697)
Topics
  • Health Sciences › Medicine › Biochemistry › Antioxidant Activity and Oxidative Stress
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Clinical Biochemistry › Paraoxonase enzyme and polymorphisms
Keywords
malondialdehyde, 4-hydroxynonenal, lipid peroxidation, lysine adducts, low-density lipoprotein, protein oxidation, Schiff base adducts, Michael addition, sodium borohydride reduction, oxidative stress biomarkers, conjugated dienes, protein cross-linking
MeSH terms
Aldehydes, Amino Acids, Collagen, Copper, Crystallins, Drug Stability, Humans, Lipoproteins, LDL, Lysine, Malondialdehyde, Gas Chromatography-Mass Spectrometry, Oxidation-Reduction, Ribonucleases, Schiff Bases, Skin, Lipid Peroxidation

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