Cross-tissue deep profiling of redox proteome in obese mice using enhanced resin-assisted capture and data-independent acquisition

Cysteine thiol modification plays a critical role in the precise regulation of the cellular redox state. Existing redox proteomic methodologies are often limited by their efficiency, complexity, and cost. To overcome these limitations, we developed and synthesized a series of novel enrichment resins. Three of these resins demonstrated a markedly enhanced enrichment efficiency. The application of one such resin, coupled with data-independent acquisition, allowed the systematic and reproducible quantification and analysis of thiol sites on a large scale within a single experiment. We further employed this strategy in a murine model of obesity and identified over 40,000 thiol sites across five tissues. These findings highlight the resin's capability to identify and quantify a significant number of thiol sites, offering a valuable resource for comprehensive investigations into redox regulation and oxidative stress in obesity.