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Plant J. 2019 Feb;97(4):765-778. doi: 10.1111/tpj.14146. Epub 2018 Dec 18.

In vivo detection of protein cysteine sulfenylation in plastids.

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Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052, Ghent, Belgium.
VIB-UGent Center for Plant Systems Biology, VIB, 9052, Ghent, Belgium.
VIB-VUB Center for Structural Biology, VIB, 1050, Brussels, Belgium.
Brussels Center for Redox Biology, 1050, Brussels, Belgium.
Structural Biology Brussels, Vrije Universiteit Brussel, 1050, Brussels, Belgium.
Department of Biochemistry, Ghent University, 9000, Ghent, Belgium.
Center for Medical Biotechnology, VIB, 9000, Ghent, Belgium.
Max Planck Institute of Molecular Plant Physiology, 14476, Potsdam-Golm, Germany.
Centre of Plant Systems Biology and Biotechnology, 4000, Plovdiv, Bulgaria.


Protein cysteine thiols are post-translationally modified under oxidative stress conditions. Illuminated chloroplasts are one of the important sources of hydrogen peroxide (H2 O2 ) and are highly sensitive to environmental stimuli, yet a comprehensive view of the oxidation-sensitive chloroplast proteome is still missing. By targeting the sulfenic acid YAP1C-trapping technology to the plastids of light-grown Arabidopsis cells, we identified 132 putatively sulfenylated plastid proteins upon H2 O2 pulse treatment. Almost half of the sulfenylated proteins are enzymes of the amino acid metabolism. Using metabolomics, we observed a reversible decrease in the levels of the amino acids Ala, Asn, Cys, Gln, Glu, His, Ile, Leu, Lys, Phe, Ser, Thr and Val after H2 O2 treatment, which is in line with an anticipated decrease in the levels of the glycolysis and tricarboxylic acid metabolites. Through the identification of an organelle-tailored proteome, we demonstrated that the subcellular targeting of the YAP1C probe enables us to study in vivo cysteine sulfenylation at the organellar level. All in all, the identification of these oxidation events in plastids revealed that several enzymes of the amino acid metabolism rapidly undergo cysteine oxidation upon oxidative stress.


Arabidopsis thaliana ; PTM ; amino acid metabolism; oxidative stress; plastid; redox signaling; sulfenic acid


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