Harnessing redox cross-reactivity to profile distinct cysteine modifications

Details

• Alternative Title:
  J Am Chem Soc
• Personal Author:
  Majmudar, Jaimeen D. ; Konopko, Aaron M. ; Labby, Kristin J. ; Tom, Christopher T.M.B. ; Crellin, John E. ; Prakash, Ashesh ; Martin, Brent R.
• Description:
  Cysteine S-nitrosation and S-sulfination are naturally occurring post-translational modifications (PTMs) on proteins induced by physiological signals and redox stress. Here we demonstrate that sulfinic acids and nitrosothiols react to form a stable thiosulfonate bond, and leverage this reactivity using sulfinate-linked probes to enrich and annotate hundreds of endogenous S-nitrosated proteins. In physiological buffers, sulfinic acids do not react with iodoacetamide or disulfides, enabling selective alkylation of free thiols and site-specific analysis of S-nitrosation. In parallel, S-nitrosothiol-linked probes enable enrichment and detection of endogenous S-sulfinated proteins, confirming that a single sulfinic acid can react with a nitrosothiol to form a thiosulfonate linkage. Using this approach, we find that hydrogen peroxide addition increases S-sulfination of human DJ-1 (PARK7) at Cys106, whereas Cys46 and Cys53 are fully oxidized to sulfonic acids. Comparative gel-based analysis of different mouse tissues reveals distinct profiles for both S-nitrosation and S-sulfination. Quantitative proteomic analysis demonstrates that both S-nitrosation and S-sulfination are widespread, yet exhibit enhanced occupancy on select proteins, including thioredoxin, peroxiredoxins, and other validated redox active proteins. Overall, we present a direct, bidirectional method to profile select redox cysteine modifications based on the unique nucleophilicity of sulfinic acids.
• Keywords:
  Cross Reactions Cysteine Humans Intracellular Signaling Peptides And Proteins Nitroso Compounds Oncogene Proteins Oxidation-Reduction Protein Deglycase DJ-1 Sulfhydryl Compounds Sulfinic Acids
• Source:
  J Am Chem Soc. 138(6):1852-1859
• Pubmed ID:
  26780921
• Pubmed Central ID:
  PMC4883004
• Document Type:
  Journal Article
• Funding:
  DP2 GM114848/DP/NCCDPHP CDC HHSUnited States/ ; F31 EB019320/EB/NIBIB NIH HHSUnited States/ ; DP2 GM114848/GM/NIGMS NIH HHSUnited States/ ; R00 CA151460/CA/NCI NIH HHSUnited States/ ; T32 CA140044/CA/NCI NIH HHSUnited States/
• Volume:
  138
• Issue:
  6
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:f24dd637aac37fac01889361ae4efeb86e46be6571f20bda024951ab1cb5a60e
• Download URL:
  https://stacks.cdc.gov/view/cdc/39896/cdc_39896_DS1.pdf
• File Type:
  [PDF - 990.22 KB ]