Identification of Anticancer ROS Targets by Cysteine Reactivity Protein Profiling

Cysteine reactivity profiling is recognized as a powerful and versatile chemoproteomic technology to study the cysteinome in native biological systems. It involves the use of chemical probes that covalently bind to reactive cysteines, enabling the enrichment and quantitively analysis of cysteine abundance through mass spectrometry. Cysteines play critical roles in maintaining cellular redox homeostasis by dynamically switching between their reduced and oxidized forms during redox perturbations. Here, we described a detailed workflow of sample preparation for cysteine-based chemical proteomics, to systematically identify cysteines that are modified by anticancer drug induced ROS.