Increased oxidation and degradation of cytosolic proteins in alcohol-exposed mouse liver and hepatoma cells
Open accessAbstract
We recently developed a sensitive method using biotin-N-maleimide (biotin-NM) as a probe to positively identify oxidized mitochondrial proteins. In this study, biotin-NM was used to identify oxidized cytosolic proteins in alcohol-fed mouse livers. Alcohol treatment for 6 wk elevated the levels of CYP2E1 and nitrotyrosine, a marker of oxidative stress. Markedly increased levels of oxidized proteins were detected in alcohol-fed mouse livers compared to pair-fed controls. The biotin-NM-labeled oxidized proteins from alcohol-exposed mouse livers were subsequently purified with streptavidin-agarose and resolved on 2-DE. More than 90 silver-stained protein spots that displayed differential intensities on 2-D gels were identified by MS. Peptide sequence analysis revealed that many enzymes or proteins involved in stress response, chaperone activity, intermediary metabolism, and antioxidant defense systems such as peroxiredoxin were oxidized after alcohol treatment. Smaller fragments of many proteins were repeatedly detected only in alcohol-fed mice, indicating that many oxidized proteins after alcohol exposure were degraded. Immunoblot results showed that the level of oxidized peroxiredoxin (inactivated) was markedly increased in the alcohol-exposed mouse livers and ethanol-sensitive hepatoma cells compared to the corresponding controls. Our results may explain the underlying mechanism for cellular dysfunction and increased susceptibility to other toxic agents following alcohol-mediated oxidative stress.
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Mass-spectral analysis revealed that more than 87 mitochondrial and 60 cytosolic proteins were oxidatively-modified in alcohol-exposed rodents.
“In fact, mass-spectral analysis revealed that more than 87 mitochondrial and 60 cytosolic proteins were oxidatively-modified in alcohol-exposed rodents [34], [35].”
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- Authors
- Kim, B., Hood, B. L., Aragon, R. A., Hardwick, J. P., Conrads, T. P., Veenstra, T. D., & Song, B. J.
- Affiliations
- National Institute on Alcohol Abuse and Alcoholism (US)
- Science Applications International Corporation (United States) (US)
- Northeast Ohio Medical University (US)
- Corresponding author
- Song, Byoung J.
- Venue
- PROTEOMICS
- Publisher
- Wiley
- Published
- 2006-02
- Volume
- 6
- Issue
- 4
- Pages
- 1250-1260
- ISSN
- 1615-9853 (ISSN-L)
- 1615-9861
- PMC
- PMC1368983
- OpenAlex
- W2159676392
- Cited by
- 83
- References
- 65
- Citations by year
- 2025: 1 · 2022: 1 · 2021: 3 · 2020: 2 · 2019: 2 · 2018: 4 · 2017: 1 · 2016: 1 · 2015: 4 · 2014: 5 · 2013: 5 · 2012: 6
- FWCI
- 4.35
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- PROTEOMICS (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- DigitalCommons-Cedarville (Cedarville University) (submittedVersion)
- Funders
- National Institutes of Health (n01co12400-79-0-22)
- National Institute on Alcohol Abuse and Alcoholism (Z01AA000036)
- National Cancer Institute (N01CO12400, N01-CO-012400)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Health Sciences › Medicine › Pathology and Forensic Medicine › Alcohol Consumption and Health Effects
- Health Sciences › Medicine › Biochemistry › Antioxidant Activity and Oxidative Stress
- Keywords
- alcohol-induced oxidative stress, protein oxidation, protein degradation, cytosolic proteins, peroxiredoxin, CYP2E1, nitrotyrosine, liver, antioxidant defense, molecular chaperones, stress response
- MeSH terms
- Ethanol, Animals, Bacterial Proteins, Biotin, Central Nervous System Depressants, Cytosol, Carcinoma, Hepatocellular, Liver, Liver Extracts, Liver Neoplasms, Male, Mice, Inbred C57BL, Oxidation-Reduction, Peroxidases, Sepharose, Biomarkers, Tumor, Tyrosine, Electrophoresis, Gel, Two-Dimensional, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Computational Biology, Cytochrome P-450 CYP2E1, Mice, Peroxiredoxins
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