Deletion of GSTA4-4 results in increased mitochondrial post-translational modification of proteins by reactive aldehydes following chronic ethanol consumption in mice
| العنوان: | Deletion of GSTA4-4 results in increased mitochondrial post-translational modification of proteins by reactive aldehydes following chronic ethanol consumption in mice |
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| المؤلفون: | Alisabeth H. Shearn, David J. Orlicky, Bridgette Engi, Kelly E. Mercer, Dennis R. Petersen, Piotr Zimniak, Colin T. Shearn, James J. Galligan, Martin J. J. Ronis, Laura Saba, Kristofer S. Fritz |
| المصدر: | Redox Biology, Vol 7, Iss C, Pp 68-77 (2016) Redox Biology |
| بيانات النشر: | Elsevier, 2016. |
| سنة النشر: | 2016 |
| مصطلحات موضوعية: | 0301 basic medicine, CID, collision-induced dissociation, Clinical Biochemistry, Mitochondrion, medicine.disease_cause, Biochemistry, ETD, electron transfer dissociation, chemistry.chemical_compound, Mice, Protein Isoforms, lcsh:QH301-705.5, Glutathione Transferase, chemistry.chemical_classification, 4-ONE, 4-oxononenal, lcsh:R5-920, 3. Good health, Amino acid, Protein carbonylation, Mitochondria, Liver, 4-HHE, 4-hydroxy-2-hexenal, lcsh:Medicine (General), EtOH, ethanol, Research Paper, ALD, alcoholic liver disease, Protein Carbonylation, Lipid peroxidation, ADPH, adipophilin, Oxidative phosphorylation, Biotin hydrazide, 4-HNE, 4-hydroxy-2-nonenal, Mitochondrial Proteins, 03 medical and health sciences, ALT, alanine aminotransferase, PF, Pair-fed, medicine, Animals, Liver Diseases, Alcoholic, MDA, malondialdehyde, Aldehydes, GSTA4, glutathione S-transferase isoform A4, Ethanol, Organic Chemistry, Metabolism, Glutathione, Molecular biology, GSTA4, Disease Models, Animal, 030104 developmental biology, Cyp2E1, Cytochrome P4502E1, chemistry, lcsh:Biology (General), Oxidative stress, Protein Processing, Post-Translational, Gene Deletion |
| الوصف: | Chronic alcohol consumption induces hepatic oxidative stress resulting in production of highly reactive electrophilic α/β-unsaturated aldehydes that have the potential to modify proteins. A primary mechanism of reactive aldehyde detoxification by hepatocytes is through GSTA4-driven enzymatic conjugation with GSH. Given reports that oxidative stress initiates GSTA4 translocation to the mitochondria, we hypothesized that increased hepatocellular damage in ethanol (EtOH)-fed GSTA4−/− mice is due to enhanced mitochondrial protein modification by reactive aldehydes. Chronic ingestion of EtOH increased hepatic protein carbonylation in GSTA4−/− mice as evidenced by increased 4-HNE and MDA immunostaining in the hepatic periportal region. Using mass spectrometric analysis of biotin hydrazide conjugated carbonylated proteins, a total of 829 proteins were identified in microsomal, cytosolic and mitochondrial fractions. Of these, 417 were novel to EtOH models. Focusing on mitochondrial fractions, 1.61-fold more carbonylated proteins were identified in EtOH-fed GSTA4−/− mice compared to their respective WT mice ingesting EtOH. Bioinformatic KEGG pathway analysis of carbonylated proteins from the mitochondrial fractions revealed an increased propensity for modification of proteins regulating oxidative phosphorylation, glucose, fatty acid, glutathione and amino acid metabolic processes in GSTA4−/− mice. Additional analysis revealed sites of reactive aldehyde protein modification on 26 novel peptides/proteins isolated from either SV/GSTA4−/− PF or EtOH fed mice. Among the peptides/proteins identified, ACSL, ACOX2, MTP, and THIKB contribute to regulation of fatty acid metabolism and ARG1, ARLY, and OAT, which regulate nitrogen and ammonia metabolism having direct relevance to ethanol-induced liver injury. These data define a role for GSTA4-4 in buffering hepatic oxidative stress associated with chronic alcohol consumption and that this GST isoform plays an important role in protecting against carbonylation of mitochondrial proteins. Graphical abstract fx1 Highlights • We demonstrate increased mitochondrial carbonylation in GSTA4-4 KO mice chronically fed EtOH. • Using LC-MS we identify 829 total carbonylated proteins (417 novel to murine ALD). • Pathway analysis revealed a propensity for adduction of fatty acid metabolic and electron transport proteins. • Using MS/MS, 26 novel adducted peptides were identified. • Reactive aldehyde modification of proteins contributes to pathogenesis of ALD. |
| اللغة: | English |
| تدمد: | 2213-2317 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::3bf0472da6658abcc58ba6a17661d04cTest http://www.sciencedirect.com/science/article/pii/S2213231715300148Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....3bf0472da6658abcc58ba6a17661d04c |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 22132317 |
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