<i>Bacillus subtilis</i> YtpP and Thioredoxin A Are New Players in the Coenzyme-A-Mediated Defense Mechanism against Cellular Stress
Coenzyme A (CoA) is an important cellular metabolite that is critical for metabolic processes and the regulation of gene expression. Recent discovery of the antioxidant function of CoA has highlighted its protective role that leads to the formation of a mixed disulfide bond with protein cysteines, w...
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| Format: | Article |
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MDPI AG
2023-04-01
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| Series: | Antioxidants |
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| Online Access: | https://www.mdpi.com/2076-3921/12/4/938 |
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| author | Maria-Armineh Tossounian Maria Baczynska William Dalton Sew Yeu Peak-Chew Kipras Undzenas George Korza Valeriy Filonenko Mark Skehel Peter Setlow Ivan Gout |
| author_facet | Maria-Armineh Tossounian Maria Baczynska William Dalton Sew Yeu Peak-Chew Kipras Undzenas George Korza Valeriy Filonenko Mark Skehel Peter Setlow Ivan Gout |
| author_sort | Maria-Armineh Tossounian |
| collection | DOAJ |
| description | Coenzyme A (CoA) is an important cellular metabolite that is critical for metabolic processes and the regulation of gene expression. Recent discovery of the antioxidant function of CoA has highlighted its protective role that leads to the formation of a mixed disulfide bond with protein cysteines, which is termed protein CoAlation. To date, more than 2000 CoAlated bacterial and mammalian proteins have been identified in cellular responses to oxidative stress, with the majority being involved in metabolic pathways (60%). Studies have shown that protein CoAlation is a widespread post-translational modification which modulates the activity and conformation of the modified proteins. The induction of protein CoAlation by oxidative stress was found to be rapidly reversed after the removal of oxidizing agents from the medium of cultured cells. In this study, we developed an enzyme-linked immunosorbent assay (ELISA)-based deCoAlation assay to detect deCoAlation activity from <i>Bacillus subtilis</i> and <i>Bacillus megaterium</i> lysates. We then used a combination of ELISA-based assay and purification strategies to show that deCoAlation is an enzyme-driven mechanism. Using mass-spectrometry and deCoAlation assays, we identified <i>B. subtilis</i> YtpP (thioredoxin-like protein) and thioredoxin A (TrxA) as enzymes that can remove CoA from different substrates. With mutagenesis studies, we identified YtpP and TrxA catalytic cysteine residues and proposed a possible deCoAlation mechanism for CoAlated methionine sulfoxide reducatse A (MsrA) and peroxiredoxin 5 (PRDX5) proteins, which results in the release of both CoA and the reduced form of MsrA or PRDX5. Overall, this paper reveals the deCoAlation activity of YtpP and TrxA and opens doors to future studies on the CoA-mediated redox regulation of CoAlated proteins under various cellular stress conditions. |
| first_indexed | 2024-03-11T05:18:18Z |
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| institution | Directory Open Access Journal |
| issn | 2076-3921 |
| language | English |
| last_indexed | 2024-03-11T05:18:18Z |
| publishDate | 2023-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antioxidants |
| spelling | doaj.art-be5094fcf3b64f328a67a25706309bd52023-11-17T18:06:32ZengMDPI AGAntioxidants2076-39212023-04-0112493810.3390/antiox12040938<i>Bacillus subtilis</i> YtpP and Thioredoxin A Are New Players in the Coenzyme-A-Mediated Defense Mechanism against Cellular StressMaria-Armineh Tossounian0Maria Baczynska1William Dalton2Sew Yeu Peak-Chew3Kipras Undzenas4George Korza5Valeriy Filonenko6Mark Skehel7Peter Setlow8Ivan Gout9Department of Structural and Molecular Biology, University College London, London WC1E 6BT, UKDepartment of Structural and Molecular Biology, University College London, London WC1E 6BT, UKDepartment of Structural and Molecular Biology, University College London, London WC1E 6BT, UKMRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Cambridge CB2 0QH, UKDepartment of Structural and Molecular Biology, University College London, London WC1E 6BT, UKDepartment of Molecular Biology and Biophysics, UConn Health, Farmington, CT 06030, USADepartment of Cell Signaling, Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, 03680 Kyiv, UkraineThe Francis Crick Institute, 1 Midland Road, London NW1 1AT, UKDepartment of Molecular Biology and Biophysics, UConn Health, Farmington, CT 06030, USADepartment of Structural and Molecular Biology, University College London, London WC1E 6BT, UKCoenzyme A (CoA) is an important cellular metabolite that is critical for metabolic processes and the regulation of gene expression. Recent discovery of the antioxidant function of CoA has highlighted its protective role that leads to the formation of a mixed disulfide bond with protein cysteines, which is termed protein CoAlation. To date, more than 2000 CoAlated bacterial and mammalian proteins have been identified in cellular responses to oxidative stress, with the majority being involved in metabolic pathways (60%). Studies have shown that protein CoAlation is a widespread post-translational modification which modulates the activity and conformation of the modified proteins. The induction of protein CoAlation by oxidative stress was found to be rapidly reversed after the removal of oxidizing agents from the medium of cultured cells. In this study, we developed an enzyme-linked immunosorbent assay (ELISA)-based deCoAlation assay to detect deCoAlation activity from <i>Bacillus subtilis</i> and <i>Bacillus megaterium</i> lysates. We then used a combination of ELISA-based assay and purification strategies to show that deCoAlation is an enzyme-driven mechanism. Using mass-spectrometry and deCoAlation assays, we identified <i>B. subtilis</i> YtpP (thioredoxin-like protein) and thioredoxin A (TrxA) as enzymes that can remove CoA from different substrates. With mutagenesis studies, we identified YtpP and TrxA catalytic cysteine residues and proposed a possible deCoAlation mechanism for CoAlated methionine sulfoxide reducatse A (MsrA) and peroxiredoxin 5 (PRDX5) proteins, which results in the release of both CoA and the reduced form of MsrA or PRDX5. Overall, this paper reveals the deCoAlation activity of YtpP and TrxA and opens doors to future studies on the CoA-mediated redox regulation of CoAlated proteins under various cellular stress conditions.https://www.mdpi.com/2076-3921/12/4/938coenzyme Aprotein CoAlationprotein deCoAlationmixed disulfideoxidative stress<i>Bacillus subtilis</i> |
| spellingShingle | Maria-Armineh Tossounian Maria Baczynska William Dalton Sew Yeu Peak-Chew Kipras Undzenas George Korza Valeriy Filonenko Mark Skehel Peter Setlow Ivan Gout <i>Bacillus subtilis</i> YtpP and Thioredoxin A Are New Players in the Coenzyme-A-Mediated Defense Mechanism against Cellular Stress Antioxidants coenzyme A protein CoAlation protein deCoAlation mixed disulfide oxidative stress <i>Bacillus subtilis</i> |
| title | <i>Bacillus subtilis</i> YtpP and Thioredoxin A Are New Players in the Coenzyme-A-Mediated Defense Mechanism against Cellular Stress |
| title_full | <i>Bacillus subtilis</i> YtpP and Thioredoxin A Are New Players in the Coenzyme-A-Mediated Defense Mechanism against Cellular Stress |
| title_fullStr | <i>Bacillus subtilis</i> YtpP and Thioredoxin A Are New Players in the Coenzyme-A-Mediated Defense Mechanism against Cellular Stress |
| title_full_unstemmed | <i>Bacillus subtilis</i> YtpP and Thioredoxin A Are New Players in the Coenzyme-A-Mediated Defense Mechanism against Cellular Stress |
| title_short | <i>Bacillus subtilis</i> YtpP and Thioredoxin A Are New Players in the Coenzyme-A-Mediated Defense Mechanism against Cellular Stress |
| title_sort | i bacillus subtilis i ytpp and thioredoxin a are new players in the coenzyme a mediated defense mechanism against cellular stress |
| topic | coenzyme A protein CoAlation protein deCoAlation mixed disulfide oxidative stress <i>Bacillus subtilis</i> |
| url | https://www.mdpi.com/2076-3921/12/4/938 |
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