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Click chemistry-based thiol redox proteomics reveals significant cysteine reduction induced by chronic ethanol consumption

In the U.S., alcohol-associated liver disease (ALD) impacts millions of people and is a major healthcare burden. While the pathology of ALD is unmistakable, the molecular mechanisms underlying ethanol hepatotoxicity are not fully understood. Hepatic ethanol metabolism is intimately linked with alter...

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Autores principales: Harris, Peter S., McGinnis, Courtney D., Michel, Cole R., Marentette, John O., Reisdorph, Richard, Roede, James R., Fritz, Kristofer S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10331594/
https://www.ncbi.nlm.nih.gov/pubmed/37390786
http://dx.doi.org/10.1016/j.redox.2023.102792
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author Harris, Peter S.
McGinnis, Courtney D.
Michel, Cole R.
Marentette, John O.
Reisdorph, Richard
Roede, James R.
Fritz, Kristofer S.
author_facet Harris, Peter S.
McGinnis, Courtney D.
Michel, Cole R.
Marentette, John O.
Reisdorph, Richard
Roede, James R.
Fritz, Kristofer S.
author_sort Harris, Peter S.
collection PubMed
description In the U.S., alcohol-associated liver disease (ALD) impacts millions of people and is a major healthcare burden. While the pathology of ALD is unmistakable, the molecular mechanisms underlying ethanol hepatotoxicity are not fully understood. Hepatic ethanol metabolism is intimately linked with alterations in extracellular and intracellular metabolic processes, specifically oxidation/reduction reactions. The xenobiotic detoxification of ethanol leads to significant disruptions in glycolysis, β-oxidation, and the TCA cycle, as well as oxidative stress. Perturbation of these regulatory networks impacts the redox status of critical regulatory protein thiols throughout the cell. Integrating these key concepts, our goal was to apply a cutting-edge approach toward understanding mechanisms of ethanol metabolism in disrupting hepatic thiol redox signaling. Utilizing a chronic murine model of ALD, we applied a cysteine targeted click chemistry enrichment coupled with quantitative nano HPLC-MS/MS to assess the thiol redox proteome. Our strategy reveals that ethanol metabolism largely reduces the cysteine proteome, with 593 cysteine residues significantly reduced and 8 significantly oxidized cysteines. Ingenuity Pathway Analysis demonstrates that ethanol metabolism reduces specific cysteines throughout ethanol metabolism (Adh1, Cat, Aldh2), antioxidant pathways (Prx1, Mgst1, Gsr), as well as many other biochemical pathways. Interestingly, a sequence motif analysis of reduced cysteines showed a correlation for hydrophilic, charged amino acids lysine or glutamic acid nearby. Further research is needed to determine how a reduced cysteine proteome impacts individual protein activity across these protein targets and pathways. Additionally, understanding how a complex array of cysteine-targeted post-translational modifications (e.g., S–NO, S-GSH, S–OH) are integrated to regulate redox signaling and control throughout the cell is key to the development of redox-centric therapeutic agents targeted to ameliorate the progression of ALD.
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spelling pubmed-103315942023-07-11 Click chemistry-based thiol redox proteomics reveals significant cysteine reduction induced by chronic ethanol consumption Harris, Peter S. McGinnis, Courtney D. Michel, Cole R. Marentette, John O. Reisdorph, Richard Roede, James R. Fritz, Kristofer S. Redox Biol Research Paper In the U.S., alcohol-associated liver disease (ALD) impacts millions of people and is a major healthcare burden. While the pathology of ALD is unmistakable, the molecular mechanisms underlying ethanol hepatotoxicity are not fully understood. Hepatic ethanol metabolism is intimately linked with alterations in extracellular and intracellular metabolic processes, specifically oxidation/reduction reactions. The xenobiotic detoxification of ethanol leads to significant disruptions in glycolysis, β-oxidation, and the TCA cycle, as well as oxidative stress. Perturbation of these regulatory networks impacts the redox status of critical regulatory protein thiols throughout the cell. Integrating these key concepts, our goal was to apply a cutting-edge approach toward understanding mechanisms of ethanol metabolism in disrupting hepatic thiol redox signaling. Utilizing a chronic murine model of ALD, we applied a cysteine targeted click chemistry enrichment coupled with quantitative nano HPLC-MS/MS to assess the thiol redox proteome. Our strategy reveals that ethanol metabolism largely reduces the cysteine proteome, with 593 cysteine residues significantly reduced and 8 significantly oxidized cysteines. Ingenuity Pathway Analysis demonstrates that ethanol metabolism reduces specific cysteines throughout ethanol metabolism (Adh1, Cat, Aldh2), antioxidant pathways (Prx1, Mgst1, Gsr), as well as many other biochemical pathways. Interestingly, a sequence motif analysis of reduced cysteines showed a correlation for hydrophilic, charged amino acids lysine or glutamic acid nearby. Further research is needed to determine how a reduced cysteine proteome impacts individual protein activity across these protein targets and pathways. Additionally, understanding how a complex array of cysteine-targeted post-translational modifications (e.g., S–NO, S-GSH, S–OH) are integrated to regulate redox signaling and control throughout the cell is key to the development of redox-centric therapeutic agents targeted to ameliorate the progression of ALD. Elsevier 2023-06-22 /pmc/articles/PMC10331594/ /pubmed/37390786 http://dx.doi.org/10.1016/j.redox.2023.102792 Text en © 2023 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research Paper
Harris, Peter S.
McGinnis, Courtney D.
Michel, Cole R.
Marentette, John O.
Reisdorph, Richard
Roede, James R.
Fritz, Kristofer S.
Click chemistry-based thiol redox proteomics reveals significant cysteine reduction induced by chronic ethanol consumption
title Click chemistry-based thiol redox proteomics reveals significant cysteine reduction induced by chronic ethanol consumption
title_full Click chemistry-based thiol redox proteomics reveals significant cysteine reduction induced by chronic ethanol consumption
title_fullStr Click chemistry-based thiol redox proteomics reveals significant cysteine reduction induced by chronic ethanol consumption
title_full_unstemmed Click chemistry-based thiol redox proteomics reveals significant cysteine reduction induced by chronic ethanol consumption
title_short Click chemistry-based thiol redox proteomics reveals significant cysteine reduction induced by chronic ethanol consumption
title_sort click chemistry-based thiol redox proteomics reveals significant cysteine reduction induced by chronic ethanol consumption
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10331594/
https://www.ncbi.nlm.nih.gov/pubmed/37390786
http://dx.doi.org/10.1016/j.redox.2023.102792
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