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Three cytosolic NAD-malate dehydrogenase isoforms of Arabidopsis thaliana: on the crossroad between energy fluxes and redox signaling

In yeast and animal cells, mitochondrial disturbances resulting from imbalances in the respiratory chain require malate dehydrogenase (MDH) activities for re-directing fluxes of reducing equivalents. In plants, in addition to mitochondria, plastids use malate valves to counterbalance and maintain re...

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Autores principales: Liszka, Aleksandra, Schimpf, Regina, Cartuche Zaruma, Krupskaya Ivannova, Buhr, Annika, Seidel, Thorsten, Walter, Stefan, Knuesting, Johannes, Dreyer, Anna, Dietz, Karl-Josef, Scheibe, Renate, Selinski, Jennifer
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7538154/
https://www.ncbi.nlm.nih.gov/pubmed/32897311
http://dx.doi.org/10.1042/BCJ20200240
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author Liszka, Aleksandra
Schimpf, Regina
Cartuche Zaruma, Krupskaya Ivannova
Buhr, Annika
Seidel, Thorsten
Walter, Stefan
Knuesting, Johannes
Dreyer, Anna
Dietz, Karl-Josef
Scheibe, Renate
Selinski, Jennifer
author_facet Liszka, Aleksandra
Schimpf, Regina
Cartuche Zaruma, Krupskaya Ivannova
Buhr, Annika
Seidel, Thorsten
Walter, Stefan
Knuesting, Johannes
Dreyer, Anna
Dietz, Karl-Josef
Scheibe, Renate
Selinski, Jennifer
author_sort Liszka, Aleksandra
collection PubMed
description In yeast and animal cells, mitochondrial disturbances resulting from imbalances in the respiratory chain require malate dehydrogenase (MDH) activities for re-directing fluxes of reducing equivalents. In plants, in addition to mitochondria, plastids use malate valves to counterbalance and maintain redox-homeostasis. Arabidopsis expresses three cytosolic MDH isoforms, namely cyMDH1, cyMDH2, and cyMDH3, the latter possessing an N-terminal extension carrying a unique cysteine residue C2. In this study, redox-effects on activity and structure of all three cyMDH isoforms were analyzed in vitro. cyMDH1 and cyMDH2 were reversibly inactivated by diamide treatment, accompanied by dimerization via disulfide-bridge formation. In contrast, cyMDH3 forms dimers and higher oligomers upon oxidation, but its low specific activity is redox-independent. In the presence of glutathione, cyMDH1 and cyMDH2 are protected from dimerization and inactivation. In contrast, cyMDH3 still dimerizes but does not form oligomers any longer. From analyses of single and double cysteine mutants and structural modeling of cyMDH3, we conclude that the presence of C2 and C336 allows for multiple cross-links in the higher molecular mass complexes comprising disulfides within the dimer as well as between monomers of two different dimers. Furthermore, nuclear localization of cyMDH isoforms was significantly increased under oxidizing conditions in isolated Arabidopsis protoplasts, in particular of isoform cyMDH3. The unique cyMDH3 C2–C2-linked dimer is, therefore, a good candidate as a redox-sensor taking over moonlighting functions upon disturbances of energy metabolism, as shown previously for the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase (GAPDH) where oxidative modification of the sensitive catalytic cysteine residues induces nuclear translocation.
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spelling pubmed-75381542020-10-15 Three cytosolic NAD-malate dehydrogenase isoforms of Arabidopsis thaliana: on the crossroad between energy fluxes and redox signaling Liszka, Aleksandra Schimpf, Regina Cartuche Zaruma, Krupskaya Ivannova Buhr, Annika Seidel, Thorsten Walter, Stefan Knuesting, Johannes Dreyer, Anna Dietz, Karl-Josef Scheibe, Renate Selinski, Jennifer Biochem J Metabolism In yeast and animal cells, mitochondrial disturbances resulting from imbalances in the respiratory chain require malate dehydrogenase (MDH) activities for re-directing fluxes of reducing equivalents. In plants, in addition to mitochondria, plastids use malate valves to counterbalance and maintain redox-homeostasis. Arabidopsis expresses three cytosolic MDH isoforms, namely cyMDH1, cyMDH2, and cyMDH3, the latter possessing an N-terminal extension carrying a unique cysteine residue C2. In this study, redox-effects on activity and structure of all three cyMDH isoforms were analyzed in vitro. cyMDH1 and cyMDH2 were reversibly inactivated by diamide treatment, accompanied by dimerization via disulfide-bridge formation. In contrast, cyMDH3 forms dimers and higher oligomers upon oxidation, but its low specific activity is redox-independent. In the presence of glutathione, cyMDH1 and cyMDH2 are protected from dimerization and inactivation. In contrast, cyMDH3 still dimerizes but does not form oligomers any longer. From analyses of single and double cysteine mutants and structural modeling of cyMDH3, we conclude that the presence of C2 and C336 allows for multiple cross-links in the higher molecular mass complexes comprising disulfides within the dimer as well as between monomers of two different dimers. Furthermore, nuclear localization of cyMDH isoforms was significantly increased under oxidizing conditions in isolated Arabidopsis protoplasts, in particular of isoform cyMDH3. The unique cyMDH3 C2–C2-linked dimer is, therefore, a good candidate as a redox-sensor taking over moonlighting functions upon disturbances of energy metabolism, as shown previously for the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase (GAPDH) where oxidative modification of the sensitive catalytic cysteine residues induces nuclear translocation. Portland Press Ltd. 2020-10-16 2020-10-05 /pmc/articles/PMC7538154/ /pubmed/32897311 http://dx.doi.org/10.1042/BCJ20200240 Text en © 2020 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle Metabolism
Liszka, Aleksandra
Schimpf, Regina
Cartuche Zaruma, Krupskaya Ivannova
Buhr, Annika
Seidel, Thorsten
Walter, Stefan
Knuesting, Johannes
Dreyer, Anna
Dietz, Karl-Josef
Scheibe, Renate
Selinski, Jennifer
Three cytosolic NAD-malate dehydrogenase isoforms of Arabidopsis thaliana: on the crossroad between energy fluxes and redox signaling
title Three cytosolic NAD-malate dehydrogenase isoforms of Arabidopsis thaliana: on the crossroad between energy fluxes and redox signaling
title_full Three cytosolic NAD-malate dehydrogenase isoforms of Arabidopsis thaliana: on the crossroad between energy fluxes and redox signaling
title_fullStr Three cytosolic NAD-malate dehydrogenase isoforms of Arabidopsis thaliana: on the crossroad between energy fluxes and redox signaling
title_full_unstemmed Three cytosolic NAD-malate dehydrogenase isoforms of Arabidopsis thaliana: on the crossroad between energy fluxes and redox signaling
title_short Three cytosolic NAD-malate dehydrogenase isoforms of Arabidopsis thaliana: on the crossroad between energy fluxes and redox signaling
title_sort three cytosolic nad-malate dehydrogenase isoforms of arabidopsis thaliana: on the crossroad between energy fluxes and redox signaling
topic Metabolism
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7538154/
https://www.ncbi.nlm.nih.gov/pubmed/32897311
http://dx.doi.org/10.1042/BCJ20200240
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