Paired-probe chemoproteomic strategy enables global site-resolved quantification of lysine redox states
- 1Sylvester Comprehensive Cancer Center. University of Miami Miller School of Medicine
Abstract
Site-specific quantification of lysine (Lys) redox states remains unexplored, as current approaches identify only a limited number of modification sites without determining their stoichiometry. Allysine, an oxidized form of Lys, provides a model posttranslational modification (PTM) for addressing this challenge by requiring selective enrichment and site-specific stoichiometric quantification. Here, we developed a quantitative chemoproteomic platform for paired redox-state isotopic stoichiometry mapping of lysine, termed PRISM-Lys. We used an Am-alkyne probe for allysine enrichment by reductive amination and developed a complementary Ald-alkyne probe to label Lys, with identical reactions generating the same chemical product. Coupling this paired-probe strategy with a quantitative chemoproteomic workflow enables global site-resolved quantification and stoichiometric analysis of Lys redox states across cells and tissues. Applied to ferroptosis, PRISM-Lys revealed bidirectional, site-selective remodeling of Lys oxidation events associated with ferroptotic progression. Together, PRISM-Lys establishes a paired-chemistry framework for global, site-resolved quantification of Lys redox states, providing a new quantitative perspective for investigating protein oxidation in biological systems.
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BibTeX
@misc{zhou2026pairedprobe,
title = {{Paired-probe chemoproteomic strategy enables global site-resolved quantification of lysine redox states}},
author = {Y.-F. Zhou and L. Zhang and D. M. N. Udayanga and X. A. WANG and H. Bakbak and H. N. Shah and N. Sharifi and Z. A. Wang},
howpublished = {bioRxiv},
year = {2026},
month = nov,
doi = {10.64898/2026.11.03.764013},
url = {https://www.biorxiv.org/content/10.64898/2026.11.03.764013v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Paired-probe chemoproteomic strategy enables global site-resolved quantification of lysine redox states
AU - Y.-F. Zhou
AU - L. Zhang
AU - D. M. N. Udayanga
AU - X. A. WANG
AU - H. Bakbak
AU - H. N. Shah
AU - N. Sharifi
AU - Z. A. Wang
PY - 2026
DA - 2026/11/03
PB - bioRxiv
DO - 10.64898/2026.11.03.764013
UR - https://www.biorxiv.org/content/10.64898/2026.11.03.764013v1
AB - Site-specific quantification of lysine (Lys) redox states remains unexplored, as current approaches identify only a limited number of modification sites without determining their stoichiometry. Allysine, an oxidized form of Lys, provides a model posttranslational modification (PTM) for addressing this challenge by requiring selective enrichment and site-specific stoichiometric quantification. Here, we developed a quantitative chemoproteomic platform for paired redox-state isotopic stoichiometry mapping of lysine, termed PRISM-Lys. We used an Am-alkyne probe for allysine enrichment by reductive amination and developed a complementary Ald-alkyne probe to label Lys, with identical reactions generating the same chemical product. Coupling this paired-probe strategy with a quantitative chemoproteomic workflow enables global site-resolved quantification and stoichiometric analysis of Lys redox states across cells and tissues. Applied to ferroptosis, PRISM-Lys revealed bidirectional, site-selective remodeling of Lys oxidation events associated with ferroptotic progression. Together, PRISM-Lys establishes a paired-chemistry framework for global, site-resolved quantification of Lys redox states, providing a new quantitative perspective for investigating protein oxidation in biological systems.
ER -