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Structural and Mechanistic Insights into C−S Bond Formation in Gliotoxin
Glutathione‐S‐transferases (GSTs) usually detoxify xenobiotics. The human pathogenic fungus Aspergillus fumigatus however uses the exceptional GST GliG to incorporate two sulfur atoms into its virulence factor gliotoxin. Because these sulfurs are essential for biological activity, glutathionylation...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8251611/ https://www.ncbi.nlm.nih.gov/pubmed/33909314 http://dx.doi.org/10.1002/anie.202104372 |
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author | Scherlach, Kirstin Kuttenlochner, Wolfgang Scharf, Daniel H. Brakhage, Axel A. Hertweck, Christian Groll, Michael Huber, Eva M. |
author_facet | Scherlach, Kirstin Kuttenlochner, Wolfgang Scharf, Daniel H. Brakhage, Axel A. Hertweck, Christian Groll, Michael Huber, Eva M. |
author_sort | Scherlach, Kirstin |
collection | PubMed |
description | Glutathione‐S‐transferases (GSTs) usually detoxify xenobiotics. The human pathogenic fungus Aspergillus fumigatus however uses the exceptional GST GliG to incorporate two sulfur atoms into its virulence factor gliotoxin. Because these sulfurs are essential for biological activity, glutathionylation is a key step of gliotoxin biosynthesis. Yet, the mechanism of carbon−sulfur linkage formation from a bis‐hydroxylated precursor is unresolved. Here, we report structures of GliG with glutathione (GSH) and its reaction product cyclo[‐l‐Phe‐l‐Ser]‐bis‐glutathione, which has been purified from a genetically modified A. fumigatus strain. The structures argue for stepwise processing of first the Phe and second the Ser moiety. Enzyme‐mediated dehydration of the substrate activates GSH and a helix dipole stabilizes the resulting anion via a water molecule for the nucleophilic attack. Activity assays with mutants validate the interactions of GliG with the ligands and enrich our knowledge about enzymatic C−S bond formation in gliotoxin and epipolythiodioxopiperazine (ETP) natural compounds in general. |
format | Online Article Text |
id | pubmed-8251611 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-82516112021-07-06 Structural and Mechanistic Insights into C−S Bond Formation in Gliotoxin Scherlach, Kirstin Kuttenlochner, Wolfgang Scharf, Daniel H. Brakhage, Axel A. Hertweck, Christian Groll, Michael Huber, Eva M. Angew Chem Int Ed Engl Research Articles Glutathione‐S‐transferases (GSTs) usually detoxify xenobiotics. The human pathogenic fungus Aspergillus fumigatus however uses the exceptional GST GliG to incorporate two sulfur atoms into its virulence factor gliotoxin. Because these sulfurs are essential for biological activity, glutathionylation is a key step of gliotoxin biosynthesis. Yet, the mechanism of carbon−sulfur linkage formation from a bis‐hydroxylated precursor is unresolved. Here, we report structures of GliG with glutathione (GSH) and its reaction product cyclo[‐l‐Phe‐l‐Ser]‐bis‐glutathione, which has been purified from a genetically modified A. fumigatus strain. The structures argue for stepwise processing of first the Phe and second the Ser moiety. Enzyme‐mediated dehydration of the substrate activates GSH and a helix dipole stabilizes the resulting anion via a water molecule for the nucleophilic attack. Activity assays with mutants validate the interactions of GliG with the ligands and enrich our knowledge about enzymatic C−S bond formation in gliotoxin and epipolythiodioxopiperazine (ETP) natural compounds in general. John Wiley and Sons Inc. 2021-05-14 2021-06-14 /pmc/articles/PMC8251611/ /pubmed/33909314 http://dx.doi.org/10.1002/anie.202104372 Text en © 2021 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | Research Articles Scherlach, Kirstin Kuttenlochner, Wolfgang Scharf, Daniel H. Brakhage, Axel A. Hertweck, Christian Groll, Michael Huber, Eva M. Structural and Mechanistic Insights into C−S Bond Formation in Gliotoxin |
title | Structural and Mechanistic Insights into C−S Bond Formation in Gliotoxin |
title_full | Structural and Mechanistic Insights into C−S Bond Formation in Gliotoxin |
title_fullStr | Structural and Mechanistic Insights into C−S Bond Formation in Gliotoxin |
title_full_unstemmed | Structural and Mechanistic Insights into C−S Bond Formation in Gliotoxin |
title_short | Structural and Mechanistic Insights into C−S Bond Formation in Gliotoxin |
title_sort | structural and mechanistic insights into c−s bond formation in gliotoxin |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8251611/ https://www.ncbi.nlm.nih.gov/pubmed/33909314 http://dx.doi.org/10.1002/anie.202104372 |
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