we developed a low-input and user-friendly pipeline for the simultaneous profiling of five distinct Cys states with regional resolution in the gut of monkeys, including free SH, total Cys oxidation (Sto), sulfenic acid (SOH), S-nitrosylation (SNO), and S-glutathionylation (SSG), shedding light on redox modification stoichiometries and redox signaling of gut aging.Furthermore, redox-modification and proteomic analysis of colonic tissues of calorie-restricted aged mice revealed that CR can reduce oxidative stress and highlight the protective effect of CR on intestinal health.