Identification of dimedone-trapped sulfenylated proteins in plants under stress
In stressed plants, the reactive oxygen species (ROS) levels rise. Key to ROS signaling research are detection and identification of the protein cysteine sulfenylation (-SOH), the ROS-mediated oxidative product of a thiol (-SH). Arabidopsis thaliana seedlings were stressed with hydrogen peroxide (H2...
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| Format: | Article |
| Language: | English |
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Elsevier
2017-03-01
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| Series: | Biochemistry and Biophysics Reports |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405580816303223 |
| _version_ | 1819087752548843520 |
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| author | Salma Akter Sebastien Carpentier Frank Van Breusegem Joris Messens |
| author_facet | Salma Akter Sebastien Carpentier Frank Van Breusegem Joris Messens |
| author_sort | Salma Akter |
| collection | DOAJ |
| description | In stressed plants, the reactive oxygen species (ROS) levels rise. Key to ROS signaling research are detection and identification of the protein cysteine sulfenylation (-SOH), the ROS-mediated oxidative product of a thiol (-SH). Arabidopsis thaliana seedlings were stressed with hydrogen peroxide (H2O2) and the sulfenylated proteins were tagged with dimedone. Dimedone-tagged sulfenic acid proteins were visualized on a two-dimensional electrophoresis (2DE) immunoblot with an anticysteine sulfenic acid antibody and were subsequently detected by mass spectrometry. We optimized the detection method for protein sulfenylation in Arabidopsis. We conclude that dimedone can penetrate the cell wall, does not stress plants, and can “read” the changes in the protein sulfenylation pattern under oxidative stress. We observed that the number of sulfenylated proteins in plants treated with 10 mM H2O2 was higher than that in untreated plants. A total of 39 sulfenylated protein spots were found on 2DE immunoblots. By means of mass spectrometry, 11 sulfenylated proteins were discovered involved in primary metabolism, redox regulation, translation and signaling pathways. Hence, by combining an immunochemical 2DE strategy with mass spectrometry, we were able to identify sulfenylated proteins in H2O2-stressed Arabidopsis seedlings. The sulfenylated proteins can be considered for further validation as redox regulators in plants. |
| first_indexed | 2024-12-21T21:41:08Z |
| format | Article |
| id | doaj.art-fb7da5c0c8874a3ea29b8d545d247766 |
| institution | Directory Open Access Journal |
| issn | 2405-5808 |
| language | English |
| last_indexed | 2024-12-21T21:41:08Z |
| publishDate | 2017-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biochemistry and Biophysics Reports |
| spelling | doaj.art-fb7da5c0c8874a3ea29b8d545d2477662022-12-21T18:49:21ZengElsevierBiochemistry and Biophysics Reports2405-58082017-03-019C10611310.1016/j.bbrep.2016.11.014Identification of dimedone-trapped sulfenylated proteins in plants under stressSalma Akter0Sebastien Carpentier1Frank Van Breusegem2Joris Messens3Department of Plant Systems Biology, VIB, 9052 Ghent, BelgiumDivision of Crop Biotechnics, KU Leuven University, 3001 Leuven, BelgiumDepartment of Plant Systems Biology, VIB, 9052 Ghent, BelgiumStructural Biology Research Center, VIB, 1050 Brussels, BelgiumIn stressed plants, the reactive oxygen species (ROS) levels rise. Key to ROS signaling research are detection and identification of the protein cysteine sulfenylation (-SOH), the ROS-mediated oxidative product of a thiol (-SH). Arabidopsis thaliana seedlings were stressed with hydrogen peroxide (H2O2) and the sulfenylated proteins were tagged with dimedone. Dimedone-tagged sulfenic acid proteins were visualized on a two-dimensional electrophoresis (2DE) immunoblot with an anticysteine sulfenic acid antibody and were subsequently detected by mass spectrometry. We optimized the detection method for protein sulfenylation in Arabidopsis. We conclude that dimedone can penetrate the cell wall, does not stress plants, and can “read” the changes in the protein sulfenylation pattern under oxidative stress. We observed that the number of sulfenylated proteins in plants treated with 10 mM H2O2 was higher than that in untreated plants. A total of 39 sulfenylated protein spots were found on 2DE immunoblots. By means of mass spectrometry, 11 sulfenylated proteins were discovered involved in primary metabolism, redox regulation, translation and signaling pathways. Hence, by combining an immunochemical 2DE strategy with mass spectrometry, we were able to identify sulfenylated proteins in H2O2-stressed Arabidopsis seedlings. The sulfenylated proteins can be considered for further validation as redox regulators in plants.http://www.sciencedirect.com/science/article/pii/S2405580816303223ArabidopsisOxidative stressSulfenylationDimedone |
| spellingShingle | Salma Akter Sebastien Carpentier Frank Van Breusegem Joris Messens Identification of dimedone-trapped sulfenylated proteins in plants under stress Biochemistry and Biophysics Reports Arabidopsis Oxidative stress Sulfenylation Dimedone |
| title | Identification of dimedone-trapped sulfenylated proteins in plants under stress |
| title_full | Identification of dimedone-trapped sulfenylated proteins in plants under stress |
| title_fullStr | Identification of dimedone-trapped sulfenylated proteins in plants under stress |
| title_full_unstemmed | Identification of dimedone-trapped sulfenylated proteins in plants under stress |
| title_short | Identification of dimedone-trapped sulfenylated proteins in plants under stress |
| title_sort | identification of dimedone trapped sulfenylated proteins in plants under stress |
| topic | Arabidopsis Oxidative stress Sulfenylation Dimedone |
| url | http://www.sciencedirect.com/science/article/pii/S2405580816303223 |
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