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Atypical Iron-Sulfur Cluster Binding, Redox Activity and Structural Properties of Chlamydomonas reinhardtii Glutaredoxin 2

Glutaredoxins (GRXs) are thioredoxin superfamily members exhibiting thiol-disulfide oxidoreductase activity and/or iron-sulfur (Fe-S) cluster binding capacities. These properties are determined by specific structural factors. In this study, we examined the capacity of the class I Chlamydomonas reinh...

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Autores principales: Roret, Thomas, Zhang, Bo, Moseler, Anna, Dhalleine, Tiphaine, Gao, Xing-Huang, Couturier, Jérémy, Lemaire, Stéphane D., Didierjean, Claude, Johnson, Michael K., Rouhier, Nicolas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8161271/
https://www.ncbi.nlm.nih.gov/pubmed/34069657
http://dx.doi.org/10.3390/antiox10050803
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author Roret, Thomas
Zhang, Bo
Moseler, Anna
Dhalleine, Tiphaine
Gao, Xing-Huang
Couturier, Jérémy
Lemaire, Stéphane D.
Didierjean, Claude
Johnson, Michael K.
Rouhier, Nicolas
author_facet Roret, Thomas
Zhang, Bo
Moseler, Anna
Dhalleine, Tiphaine
Gao, Xing-Huang
Couturier, Jérémy
Lemaire, Stéphane D.
Didierjean, Claude
Johnson, Michael K.
Rouhier, Nicolas
author_sort Roret, Thomas
collection PubMed
description Glutaredoxins (GRXs) are thioredoxin superfamily members exhibiting thiol-disulfide oxidoreductase activity and/or iron-sulfur (Fe-S) cluster binding capacities. These properties are determined by specific structural factors. In this study, we examined the capacity of the class I Chlamydomonas reinhardtii GRX2 recombinant protein to catalyze both protein glutathionylation and deglutathionylation reactions using a redox sensitive fluorescent protein as a model protein substrate. We observed that the catalytic cysteine of the CPYC active site motif of GRX2 was sufficient for catalyzing both reactions in the presence of glutathione. Unexpectedly, spectroscopic characterization of the protein purified under anaerobiosis showed the presence of a [2Fe-2S] cluster despite having a presumably inadequate active site signature, based on past mutational analyses. The spectroscopic characterization of cysteine mutated variants together with modeling of the Fe–S cluster-bound GRX homodimer from the structure of an apo-GRX2 indicate the existence of an atypical Fe–S cluster environment and ligation mode. Overall, the results further delineate the biochemical and structural properties of conventional GRXs, pointing to the existence of multiple factors more complex than anticipated, sustaining the capacity of these proteins to bind Fe–S clusters.
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spelling pubmed-81612712021-05-29 Atypical Iron-Sulfur Cluster Binding, Redox Activity and Structural Properties of Chlamydomonas reinhardtii Glutaredoxin 2 Roret, Thomas Zhang, Bo Moseler, Anna Dhalleine, Tiphaine Gao, Xing-Huang Couturier, Jérémy Lemaire, Stéphane D. Didierjean, Claude Johnson, Michael K. Rouhier, Nicolas Antioxidants (Basel) Article Glutaredoxins (GRXs) are thioredoxin superfamily members exhibiting thiol-disulfide oxidoreductase activity and/or iron-sulfur (Fe-S) cluster binding capacities. These properties are determined by specific structural factors. In this study, we examined the capacity of the class I Chlamydomonas reinhardtii GRX2 recombinant protein to catalyze both protein glutathionylation and deglutathionylation reactions using a redox sensitive fluorescent protein as a model protein substrate. We observed that the catalytic cysteine of the CPYC active site motif of GRX2 was sufficient for catalyzing both reactions in the presence of glutathione. Unexpectedly, spectroscopic characterization of the protein purified under anaerobiosis showed the presence of a [2Fe-2S] cluster despite having a presumably inadequate active site signature, based on past mutational analyses. The spectroscopic characterization of cysteine mutated variants together with modeling of the Fe–S cluster-bound GRX homodimer from the structure of an apo-GRX2 indicate the existence of an atypical Fe–S cluster environment and ligation mode. Overall, the results further delineate the biochemical and structural properties of conventional GRXs, pointing to the existence of multiple factors more complex than anticipated, sustaining the capacity of these proteins to bind Fe–S clusters. MDPI 2021-05-19 /pmc/articles/PMC8161271/ /pubmed/34069657 http://dx.doi.org/10.3390/antiox10050803 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Roret, Thomas
Zhang, Bo
Moseler, Anna
Dhalleine, Tiphaine
Gao, Xing-Huang
Couturier, Jérémy
Lemaire, Stéphane D.
Didierjean, Claude
Johnson, Michael K.
Rouhier, Nicolas
Atypical Iron-Sulfur Cluster Binding, Redox Activity and Structural Properties of Chlamydomonas reinhardtii Glutaredoxin 2
title Atypical Iron-Sulfur Cluster Binding, Redox Activity and Structural Properties of Chlamydomonas reinhardtii Glutaredoxin 2
title_full Atypical Iron-Sulfur Cluster Binding, Redox Activity and Structural Properties of Chlamydomonas reinhardtii Glutaredoxin 2
title_fullStr Atypical Iron-Sulfur Cluster Binding, Redox Activity and Structural Properties of Chlamydomonas reinhardtii Glutaredoxin 2
title_full_unstemmed Atypical Iron-Sulfur Cluster Binding, Redox Activity and Structural Properties of Chlamydomonas reinhardtii Glutaredoxin 2
title_short Atypical Iron-Sulfur Cluster Binding, Redox Activity and Structural Properties of Chlamydomonas reinhardtii Glutaredoxin 2
title_sort atypical iron-sulfur cluster binding, redox activity and structural properties of chlamydomonas reinhardtii glutaredoxin 2
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8161271/
https://www.ncbi.nlm.nih.gov/pubmed/34069657
http://dx.doi.org/10.3390/antiox10050803
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