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Redox Conformation-Specific Protein–Protein Interactions of the 2-Cysteine Peroxiredoxin in Arabidopsis
2-Cysteine peroxiredoxins (2-CysPRX) are highly abundant thiol peroxidases in chloroplasts and play key roles in reactive oxygen species (ROS) defense and redox signaling. Peroxide-dependent oxidation of cysteines induces conformational changes that alter the ability for protein–protein interactions...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7346168/ https://www.ncbi.nlm.nih.gov/pubmed/32545358 http://dx.doi.org/10.3390/antiox9060515 |
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author | Liebthal, Michael Schuetze, Johannes Dreyer, Anna Mock, Hans-Peter Dietz, Karl-Josef |
author_facet | Liebthal, Michael Schuetze, Johannes Dreyer, Anna Mock, Hans-Peter Dietz, Karl-Josef |
author_sort | Liebthal, Michael |
collection | PubMed |
description | 2-Cysteine peroxiredoxins (2-CysPRX) are highly abundant thiol peroxidases in chloroplasts and play key roles in reactive oxygen species (ROS) defense and redox signaling. Peroxide-dependent oxidation of cysteines induces conformational changes that alter the ability for protein–protein interactions. For regeneration, 2-CysPRXs withdraw electrons from thioredoxins (TRXs) and participate in redox-dependent regulation by affecting the redox state of TRX-dependent targets, for example, in chloroplast metabolism. This work explores the redox conformation-specific 2-CysPRX interactome using an affinity-based pull down with recombinant variants arrested in specific quaternary conformations. This allowed us to address a critical and poorly explored aspect of the redox-regulatory network and showed that the interaction of TRXs, their interaction partners, and 2-CysPRX occur under contrasting redox conditions. A set of 178 chloroplast proteins were identified from leaf proteins and included proteins with functions in photosynthesis, carbohydrate, fatty acid and amino acid metabolism, and defense. These processes are known to be deregulated in plants devoid of 2-CysPRX. Selected enzymes like LIPOXYGENASE 2, CHLOROPLAST PROTEIN 12-1, CHORISMATE SYNTHASE, ß-CARBONIC ANHYDRASE, and FERREDOXIN-dependent GLUTAMATE SYNTHASE 1 were subjected to far Western, isothermal titration calorimetry, and enzyme assays for validation. The pull down fractions frequently contained TRXs as well as their target proteins, for example, FRUCTOSE-1,6-BISPHOSPHATASE and MALATE DEHYDROGENASE. The difference between TRX-dependent indirect interactions of TRX targets and 2-CysPRX and direct 2-CysPRX binding is hypothesized to be related to quaternary structure formation, where 2-CysPRX oligomers function as scaffold for complex formation, whereas TRX oxidase activity of 2-CysPRX controls the redox state of TRX-related enzyme activity. |
format | Online Article Text |
id | pubmed-7346168 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-73461682020-07-14 Redox Conformation-Specific Protein–Protein Interactions of the 2-Cysteine Peroxiredoxin in Arabidopsis Liebthal, Michael Schuetze, Johannes Dreyer, Anna Mock, Hans-Peter Dietz, Karl-Josef Antioxidants (Basel) Article 2-Cysteine peroxiredoxins (2-CysPRX) are highly abundant thiol peroxidases in chloroplasts and play key roles in reactive oxygen species (ROS) defense and redox signaling. Peroxide-dependent oxidation of cysteines induces conformational changes that alter the ability for protein–protein interactions. For regeneration, 2-CysPRXs withdraw electrons from thioredoxins (TRXs) and participate in redox-dependent regulation by affecting the redox state of TRX-dependent targets, for example, in chloroplast metabolism. This work explores the redox conformation-specific 2-CysPRX interactome using an affinity-based pull down with recombinant variants arrested in specific quaternary conformations. This allowed us to address a critical and poorly explored aspect of the redox-regulatory network and showed that the interaction of TRXs, their interaction partners, and 2-CysPRX occur under contrasting redox conditions. A set of 178 chloroplast proteins were identified from leaf proteins and included proteins with functions in photosynthesis, carbohydrate, fatty acid and amino acid metabolism, and defense. These processes are known to be deregulated in plants devoid of 2-CysPRX. Selected enzymes like LIPOXYGENASE 2, CHLOROPLAST PROTEIN 12-1, CHORISMATE SYNTHASE, ß-CARBONIC ANHYDRASE, and FERREDOXIN-dependent GLUTAMATE SYNTHASE 1 were subjected to far Western, isothermal titration calorimetry, and enzyme assays for validation. The pull down fractions frequently contained TRXs as well as their target proteins, for example, FRUCTOSE-1,6-BISPHOSPHATASE and MALATE DEHYDROGENASE. The difference between TRX-dependent indirect interactions of TRX targets and 2-CysPRX and direct 2-CysPRX binding is hypothesized to be related to quaternary structure formation, where 2-CysPRX oligomers function as scaffold for complex formation, whereas TRX oxidase activity of 2-CysPRX controls the redox state of TRX-related enzyme activity. MDPI 2020-06-11 /pmc/articles/PMC7346168/ /pubmed/32545358 http://dx.doi.org/10.3390/antiox9060515 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Liebthal, Michael Schuetze, Johannes Dreyer, Anna Mock, Hans-Peter Dietz, Karl-Josef Redox Conformation-Specific Protein–Protein Interactions of the 2-Cysteine Peroxiredoxin in Arabidopsis |
title | Redox Conformation-Specific Protein–Protein Interactions of the 2-Cysteine Peroxiredoxin in Arabidopsis |
title_full | Redox Conformation-Specific Protein–Protein Interactions of the 2-Cysteine Peroxiredoxin in Arabidopsis |
title_fullStr | Redox Conformation-Specific Protein–Protein Interactions of the 2-Cysteine Peroxiredoxin in Arabidopsis |
title_full_unstemmed | Redox Conformation-Specific Protein–Protein Interactions of the 2-Cysteine Peroxiredoxin in Arabidopsis |
title_short | Redox Conformation-Specific Protein–Protein Interactions of the 2-Cysteine Peroxiredoxin in Arabidopsis |
title_sort | redox conformation-specific protein–protein interactions of the 2-cysteine peroxiredoxin in arabidopsis |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7346168/ https://www.ncbi.nlm.nih.gov/pubmed/32545358 http://dx.doi.org/10.3390/antiox9060515 |
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