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Identification of sulfenylated cysteines in Arabidopsis thaliana proteins using a disulfide-linked peptide reporter

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Abstract
In proteins, hydrogen peroxide (H2O2) reacts with redox-sensitive cysteines to form cysteine sulfenic acid, also known asS-sulfenylation. These cysteine oxidation events can steer diverse cellular processes by altering protein interactions, trafficking, conformation, and function. Previously, we had identifiedS-sulfenylated proteins by using a tagged proteinaceous probe based on the yeast AP-1-like (Yap1) transcription factor that specifically reacts with sulfenic acids and traps them through a mixed disulfide bond. However, the identity of theS-sulfenylated amino acid residues within a protein remained enigmatic. By using the same transgenic YAP1C probe, we present here a technological advancement to identifyin situsulfenylated cysteine sites inArabidopsis thalianacells under control condition and oxidative stress. Briefly, the total extract of transgenic YAP1CA. thalianacells was initially purified on IgG-Sepharose beads, followed by a tryptic digest. Then, the mixed disulfide-linked peptides were further enriched at the peptide level on an anti-YAP1C-derived peptide (C598SEIWDR) antibody. Subsequent mass spectrometry analysis with pLink 2 identified 1,745 YAP1C cross-linked peptides, indicating sulfenylated cysteines in over 1,000 proteins. Approximately 55% of these YAP1C-linked cysteines had previously been reported as redox-sensitive cysteines (S-sulfenylation,S-nitrosylation, and reversibly oxidized cysteines). The presented methodology provides a noninvasive approach to identify sulfenylated cysteines in any species that can be genetically modified.
Keywords
Plant Science, S-sulfenylation (-SOH), YAP1C, hydrogen peroxide, cross-linked peptide identification, affinity purification, disulfide, Arabidopsis thaliana, GENETICALLY ENCODED PROBE, S-NITROSYLATED PROTEINS, SULFENIC ACID, PURIFICATION, PEROXIDASE, RECEPTOR, STRESS, H2O2

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MLA
Wei, Bo, et al. “Identification of Sulfenylated Cysteines in Arabidopsis Thaliana Proteins Using a Disulfide-Linked Peptide Reporter.” FRONTIERS IN PLANT SCIENCE, vol. 11, 2020, doi:10.3389/fpls.2020.00777.
APA
Wei, B., Willems, P., Huang, J., Tian, C., Yang, J., Messens, J., & Van Breusegem, F. (2020). Identification of sulfenylated cysteines in Arabidopsis thaliana proteins using a disulfide-linked peptide reporter. FRONTIERS IN PLANT SCIENCE, 11. https://doi.org/10.3389/fpls.2020.00777
Chicago author-date
Wei, Bo, Patrick Willems, Jingjing Huang, Caiping Tian, Jing Yang, Joris Messens, and Frank Van Breusegem. 2020. “Identification of Sulfenylated Cysteines in Arabidopsis Thaliana Proteins Using a Disulfide-Linked Peptide Reporter.” FRONTIERS IN PLANT SCIENCE 11. https://doi.org/10.3389/fpls.2020.00777.
Chicago author-date (all authors)
Wei, Bo, Patrick Willems, Jingjing Huang, Caiping Tian, Jing Yang, Joris Messens, and Frank Van Breusegem. 2020. “Identification of Sulfenylated Cysteines in Arabidopsis Thaliana Proteins Using a Disulfide-Linked Peptide Reporter.” FRONTIERS IN PLANT SCIENCE 11. doi:10.3389/fpls.2020.00777.
Vancouver
1.
Wei B, Willems P, Huang J, Tian C, Yang J, Messens J, et al. Identification of sulfenylated cysteines in Arabidopsis thaliana proteins using a disulfide-linked peptide reporter. FRONTIERS IN PLANT SCIENCE. 2020;11.
IEEE
[1]
B. Wei et al., “Identification of sulfenylated cysteines in Arabidopsis thaliana proteins using a disulfide-linked peptide reporter,” FRONTIERS IN PLANT SCIENCE, vol. 11, 2020.
@article{8670654,
  abstract     = {{In proteins, hydrogen peroxide (H2O2) reacts with redox-sensitive cysteines to form cysteine sulfenic acid, also known asS-sulfenylation. These cysteine oxidation events can steer diverse cellular processes by altering protein interactions, trafficking, conformation, and function. Previously, we had identifiedS-sulfenylated proteins by using a tagged proteinaceous probe based on the yeast AP-1-like (Yap1) transcription factor that specifically reacts with sulfenic acids and traps them through a mixed disulfide bond. However, the identity of theS-sulfenylated amino acid residues within a protein remained enigmatic. By using the same transgenic YAP1C probe, we present here a technological advancement to identifyin situsulfenylated cysteine sites inArabidopsis thalianacells under control condition and oxidative stress. Briefly, the total extract of transgenic YAP1CA. thalianacells was initially purified on IgG-Sepharose beads, followed by a tryptic digest. Then, the mixed disulfide-linked peptides were further enriched at the peptide level on an anti-YAP1C-derived peptide (C598SEIWDR) antibody. Subsequent mass spectrometry analysis with pLink 2 identified 1,745 YAP1C cross-linked peptides, indicating sulfenylated cysteines in over 1,000 proteins. Approximately 55% of these YAP1C-linked cysteines had previously been reported as redox-sensitive cysteines (S-sulfenylation,S-nitrosylation, and reversibly oxidized cysteines). The presented methodology provides a noninvasive approach to identify sulfenylated cysteines in any species that can be genetically modified.}},
  articleno    = {{777}},
  author       = {{Wei, Bo and Willems, Patrick and Huang, Jingjing and Tian, Caiping and Yang, Jing and Messens, Joris and Van Breusegem, Frank}},
  issn         = {{1664-462X}},
  journal      = {{FRONTIERS IN PLANT SCIENCE}},
  keywords     = {{Plant Science,S-sulfenylation (-SOH),YAP1C,hydrogen peroxide,cross-linked peptide identification,affinity purification,disulfide,Arabidopsis thaliana,GENETICALLY ENCODED PROBE,S-NITROSYLATED PROTEINS,SULFENIC ACID,PURIFICATION,PEROXIDASE,RECEPTOR,STRESS,H2O2}},
  language     = {{eng}},
  pages        = {{12}},
  title        = {{Identification of sulfenylated cysteines in Arabidopsis thaliana proteins using a disulfide-linked peptide reporter}},
  url          = {{http://dx.doi.org/10.3389/fpls.2020.00777}},
  volume       = {{11}},
  year         = {{2020}},
}

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