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Cited by 19Alcohol Consumption and Health EffectsAntioxidant Activity and Oxidative StressRedox biology and oxidative stress

Increased hepatocellular protein carbonylation in human end-stage alcoholic cirrhosis

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Shearn, C.T., Orlicky, D.J., Saba, L.M., Shearn, A.H., Petersen, Dennis R.·2015·Free Radical Biology and Medicine
DOI 10.1016/j.freeradbiomed.2015.10.420PMC PMC4762037
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Abstract

Objective Oxidative stress is a significant contributing factor in the pathogenesis of alcoholic liver disease (ALD). In the murine models of chronic alcohol consumption, induction of oxidative stress results in increased peroxidation of polyunsaturated fatty acids to form highly reactive electrophilic α/β unsaturated aldehydes that post-translationally modify proteins altering activity. Data are presented here suggesting that oxidative stress and the resulting carbonylation of hepatic proteins is an ongoing process involved in alcohol-induced cirrhosis. Methods Using age-matched pooled hepatic tissue obtained from healthy humans and patients with end stage cirrhotic ALD, overall carbonylation was assessed by immunohistochemistry and LC-MS/MS of streptavidin purified hepatic whole cell extracts treated with biotin hydrazide. Identified carbonylated proteins were further evaluated using bioinformatics analyses. Results Using immunohistochemistry and Western blotting, protein carbonylation was increased in end stage ALD occurring primarily in hepatocytes. Mass spectrometric analysis revealed a total of 1224 carbonylated proteins in normal hepatic and end-stage alcoholic cirrhosis tissue. Of these, 411 were unique to cirrhotic ALD, 261 unique to normal hepatic tissue and 552 common to both groups. Bioinformatic pathway analysis of hepatic carbonylated proteins revealed a propensity of long term EtOH consumption to increase post-translational carbonylation of proteins involved in glutathione homeostatic, glycolytic and cytoskeletal pathways. Western analysis revealed increased expression of GSTA4 and GSTπ in human ALD. Using LC-MS/MS analysis, a nonenaldehyde post-translational modification was identified on Lysine 235 of the cytoskeletal protein vimentin in whole cell extracts prepared from human end stage ALD hepatic tissue. Conclusions These studies are the first to use LC-MS/MS analysis of carbonylated proteins in human ALD and begin exploring possible mechanistic links with end-stage alcoholic cirrhosis and oxidative stress.

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Authors
Shearn, C. T., Orlicky, D. J., Saba, L. M., Shearn, A. H., & Petersen, D. R.
Affiliations
University of Colorado Anschutz (US)
Corresponding author
  • Shearn, C.T.
  • Petersen, Dennis R.
ORCID
Venue
Free Radical Biology and Medicine
Publisher
Elsevier BV
Published
2015-12
Volume
89
Pages
1144-1153
ISSN
0891-5849 (ISSN-L)
DOI
10.1016/j.freeradbiomed.2015.10.420
PMC
PMC4762037
OpenAlex
W1869371862
Cited by
19
References
74
Citations by year
2026: 1 · 2025: 2 · 2024: 3 · 2021: 1 · 2020: 3 · 2019: 1 · 2018: 4 · 2017: 3 · 2016: 1
FWCI
2.12
Type
journal article
Language
en
Open access
Licence
Locations
  • Free Radical Biology and Medicine (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
National Institute on Alcohol Abuse and Alcoholism (5R37 AA009300-18)
Topics
  • Health Sciences › Medicine › Pathology and Forensic Medicine › Alcohol Consumption and Health Effects
  • Health Sciences › Medicine › Biochemistry › Antioxidant Activity and Oxidative Stress
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
Keywords
protein carbonylation, alcoholic liver disease, oxidative stress, hepatocytes, glutathione S-transferase pi, vimentin, post-translational modification, lipid peroxidation, glutathione homeostasis, glycolysis, cytoskeleton
MeSH terms
Adult, Chromatography, Liquid, Female, Humans, Liver Cirrhosis, Alcoholic, Male, Middle Aged, Protein Processing, Post-Translational, Blotting, Western, Biomarkers, Case-Control Studies, Protein Carbonylation, Tandem Mass Spectrometry

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