The dithiol mechanism of class I glutaredoxins promotes specificity for glutathione as a reducing agent
Class I glutaredoxins reversibly reduce glutathione- and nonglutathione disulfides with the help of reduced glutathione (GSH) using either a monothiol mechanism or a dithiol mechanism. The monothiol mechanism exclusively involves a single glutathionylated active-site cysteinyl residue, whereas the d...
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Elsevier
2024-12-01
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| Series: | Redox Biology |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213231724003884 |
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| author | Lukas Lang Philipp Reinert Cedric Diaz Marcel Deponte |
| author_facet | Lukas Lang Philipp Reinert Cedric Diaz Marcel Deponte |
| author_sort | Lukas Lang |
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| description | Class I glutaredoxins reversibly reduce glutathione- and nonglutathione disulfides with the help of reduced glutathione (GSH) using either a monothiol mechanism or a dithiol mechanism. The monothiol mechanism exclusively involves a single glutathionylated active-site cysteinyl residue, whereas the dithiol mechanism requires the additional formation of an intramolecular disulfide bond between the active-site cysteinyl residue and a resolving cysteinyl residue. While the oxidation of glutaredoxins by glutathione disulfide substrates has been extensively characterized, the enzyme-substrate interactions for the reduction of S-glutathionylated glutaredoxins or intramolecular glutaredoxin disulfides are still poorly characterized. Here we compared the thiol-specificity for the reduction of S-glutathionylated glutaredoxins and the intramolecular glutaredoxin disulfide. We show that S-glutathionylated glutaredoxins rapidly react with a plethora of thiols and that the 2nd glutathione-interaction site of class I glutaredoxins lacks specificity for GSH as a reducing agent. In contrast, the slower reduction of the partially strained intramolecular glutaredoxin disulfide involves specific interactions with both carboxylate groups of GSH at the 1st glutathione-interaction site. Thus, the dithiol mechanism of class I glutaredoxins promotes specificity for GSH as a reducing agent, which might explain the prevalence of dithiol glutaredoxins in pro- and eukaryotes. |
| format | Article |
| id | doaj-art-df2f1c91a17943b78f6635b9d77bc4bb |
| institution | Kabale University |
| issn | 2213-2317 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Redox Biology |
| spelling | doaj-art-df2f1c91a17943b78f6635b9d77bc4bb2024-12-08T06:09:57ZengElsevierRedox Biology2213-23172024-12-0178103410The dithiol mechanism of class I glutaredoxins promotes specificity for glutathione as a reducing agentLukas Lang0Philipp Reinert1Cedric Diaz2Marcel Deponte3Faculty of Chemistry, Comparative Biochemistry, RPTU Kaiserslautern, D-67663, Kaiserslautern, GermanyFaculty of Chemistry, Comparative Biochemistry, RPTU Kaiserslautern, D-67663, Kaiserslautern, GermanyFaculty of Chemistry, Comparative Biochemistry, RPTU Kaiserslautern, D-67663, Kaiserslautern, GermanyCorresponding author.; Faculty of Chemistry, Comparative Biochemistry, RPTU Kaiserslautern, D-67663, Kaiserslautern, GermanyClass I glutaredoxins reversibly reduce glutathione- and nonglutathione disulfides with the help of reduced glutathione (GSH) using either a monothiol mechanism or a dithiol mechanism. The monothiol mechanism exclusively involves a single glutathionylated active-site cysteinyl residue, whereas the dithiol mechanism requires the additional formation of an intramolecular disulfide bond between the active-site cysteinyl residue and a resolving cysteinyl residue. While the oxidation of glutaredoxins by glutathione disulfide substrates has been extensively characterized, the enzyme-substrate interactions for the reduction of S-glutathionylated glutaredoxins or intramolecular glutaredoxin disulfides are still poorly characterized. Here we compared the thiol-specificity for the reduction of S-glutathionylated glutaredoxins and the intramolecular glutaredoxin disulfide. We show that S-glutathionylated glutaredoxins rapidly react with a plethora of thiols and that the 2nd glutathione-interaction site of class I glutaredoxins lacks specificity for GSH as a reducing agent. In contrast, the slower reduction of the partially strained intramolecular glutaredoxin disulfide involves specific interactions with both carboxylate groups of GSH at the 1st glutathione-interaction site. Thus, the dithiol mechanism of class I glutaredoxins promotes specificity for GSH as a reducing agent, which might explain the prevalence of dithiol glutaredoxins in pro- and eukaryotes.http://www.sciencedirect.com/science/article/pii/S2213231724003884Enzyme mechanismDisulfideDithiolGlutaredoxinGlutathioneRedox catalysis |
| spellingShingle | Lukas Lang Philipp Reinert Cedric Diaz Marcel Deponte The dithiol mechanism of class I glutaredoxins promotes specificity for glutathione as a reducing agent Redox Biology Enzyme mechanism Disulfide Dithiol Glutaredoxin Glutathione Redox catalysis |
| title | The dithiol mechanism of class I glutaredoxins promotes specificity for glutathione as a reducing agent |
| title_full | The dithiol mechanism of class I glutaredoxins promotes specificity for glutathione as a reducing agent |
| title_fullStr | The dithiol mechanism of class I glutaredoxins promotes specificity for glutathione as a reducing agent |
| title_full_unstemmed | The dithiol mechanism of class I glutaredoxins promotes specificity for glutathione as a reducing agent |
| title_short | The dithiol mechanism of class I glutaredoxins promotes specificity for glutathione as a reducing agent |
| title_sort | dithiol mechanism of class i glutaredoxins promotes specificity for glutathione as a reducing agent |
| topic | Enzyme mechanism Disulfide Dithiol Glutaredoxin Glutathione Redox catalysis |
| url | http://www.sciencedirect.com/science/article/pii/S2213231724003884 |
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