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Disulfide bond formation protects Arabidopsis thaliana glutathione transferase tau 23 from oxidative damage

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Biotechnology for a sustainable economy (Bio-Economy)
Abstract
Background: Glutathione transferases play an important role as detoxifying enzymes. In A. thaliana, elevated levels of reactive oxygen species (ROS), provoked during biotic and abiotic stress, influence the activity of GSTU23. The aim of this study is to determine the impact of oxidative stress on the function and structure of GSTU23. Methods: The impact of oxidation on the function of GSTU23 was studied using a glutathione transferase biochemical assay and mass spectrometry. With kinetics, circular dichroism and thermodynamics, we compared reduced with oxidized GSTU23. X-ray crystal structures of GSTU23 visualize the impact of oxidation on methionines and cysteines. Results: In the presence of 100 mu M H2O2, oxidation of the methionine side-chain to a sulfoxide is the prominent post-translational modification, which can be reduced by C. diphtheriae MsrA and MsrB. However, increasing the level to 200 mu M H2O2 results in a reversible intramolecular disulfide between Cys65-Cys110, which is substrate for glutaredoxin. Under these oxidizing conditions, GSTU23 undergoes a structural change and forms a more favourable enzyme-substrate complex to overcome k(cat) decrease. Conclusions and significance: At lower H2O2 levels (100 mu M), GSTU23 forms methionine sulfoxides. Specifically, oxidation of Met14, located near the catalytic Ser13, could interfere with both GSH binding and catalytic activation. At higher H2O2 levels (200 mu M), the Cys65-Cys110 disulfide bond protects other cysteines and also methionines from overoxidation. This study shows the impact of oxidative stress on GSTU23 regulated by methionine sulfoxide reductases and glutaredoxin, and the mechanisms involved in maintaining its catalytic functionality under oxidizing conditions.
Keywords
METHIONINE SULFOXIDE REDUCTASE, ACTIVE-SITE ARCHITECTURE, S-TRANSFERASE, CYSTEINE DIOXYGENASE, ESCHERICHIA-COLI, REDOX REGULATION, ACID, FORMATION, GENE FAMILY, STRESS, MECHANISM, Glutathione transferase, Kinetics, X-ray structure, Thermodynamics, Methionine sulfoxide, Disulfide bond

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Citation

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Chicago
Tossounian, Maria-Armineh, Inge Van Molle, Khadija Wahni, Silke Jacques, Kris Gevaert, Frank Van Breusegem, Didier Vertommen, David Young, Leonardo Astolfi Rosado, and Joris Messens. 2018. “Disulfide Bond Formation Protects Arabidopsis Thaliana Glutathione Transferase Tau 23 from Oxidative Damage.” Biochimica Et Biophysica Acta-general Subjects 1862 (3): 775–789.
APA
Tossounian, M.-A., Van Molle, I., Wahni, K., Jacques, S., Gevaert, K., Van Breusegem, F., Vertommen, D., et al. (2018). Disulfide bond formation protects Arabidopsis thaliana glutathione transferase tau 23 from oxidative damage. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, 1862(3), 775–789.
Vancouver
1.
Tossounian M-A, Van Molle I, Wahni K, Jacques S, Gevaert K, Van Breusegem F, et al. Disulfide bond formation protects Arabidopsis thaliana glutathione transferase tau 23 from oxidative damage. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS. 2018;1862(3):775–89.
MLA
Tossounian, Maria-Armineh, Inge Van Molle, Khadija Wahni, et al. “Disulfide Bond Formation Protects Arabidopsis Thaliana Glutathione Transferase Tau 23 from Oxidative Damage.” BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS 1862.3 (2018): 775–789. Print.
@article{8558154,
  abstract     = {Background: Glutathione transferases play an important role as detoxifying enzymes. In A. thaliana, elevated levels of reactive oxygen species (ROS), provoked during biotic and abiotic stress, influence the activity of GSTU23. The aim of this study is to determine the impact of oxidative stress on the function and structure of GSTU23. 
Methods: The impact of oxidation on the function of GSTU23 was studied using a glutathione transferase biochemical assay and mass spectrometry. With kinetics, circular dichroism and thermodynamics, we compared reduced with oxidized GSTU23. X-ray crystal structures of GSTU23 visualize the impact of oxidation on methionines and cysteines. 
Results: In the presence of 100 mu M H2O2, oxidation of the methionine side-chain to a sulfoxide is the prominent post-translational modification, which can be reduced by C. diphtheriae MsrA and MsrB. However, increasing the level to 200 mu M H2O2 results in a reversible intramolecular disulfide between Cys65-Cys110, which is substrate for glutaredoxin. Under these oxidizing conditions, GSTU23 undergoes a structural change and forms a more favourable enzyme-substrate complex to overcome k(cat) decrease. 
Conclusions and significance: At lower H2O2 levels (100 mu M), GSTU23 forms methionine sulfoxides. Specifically, oxidation of Met14, located near the catalytic Ser13, could interfere with both GSH binding and catalytic activation. At higher H2O2 levels (200 mu M), the Cys65-Cys110 disulfide bond protects other cysteines and also methionines from overoxidation. This study shows the impact of oxidative stress on GSTU23 regulated by methionine sulfoxide reductases and glutaredoxin, and the mechanisms involved in maintaining its catalytic functionality under oxidizing conditions.},
  author       = {Tossounian, Maria-Armineh and Van Molle, Inge and Wahni, Khadija and Jacques, Silke and Gevaert, Kris and Van Breusegem, Frank and Vertommen, Didier and Young, David and Rosado, Leonardo Astolfi and Messens, Joris},
  issn         = {0304-4165},
  journal      = {BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS},
  keyword      = {METHIONINE SULFOXIDE REDUCTASE,ACTIVE-SITE ARCHITECTURE,S-TRANSFERASE,CYSTEINE DIOXYGENASE,ESCHERICHIA-COLI,REDOX REGULATION,ACID,FORMATION,GENE FAMILY,STRESS,MECHANISM,Glutathione transferase,Kinetics,X-ray structure,Thermodynamics,Methionine sulfoxide,Disulfide bond},
  language     = {eng},
  number       = {3},
  pages        = {775--789},
  title        = {Disulfide bond formation protects Arabidopsis thaliana glutathione transferase tau 23 from oxidative damage},
  url          = {http://dx.doi.org/10.1016/j.bbagen.2017.10.007},
  volume       = {1862},
  year         = {2018},
}

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