Regular physical activity elicits broad physiological adaptations, yet the molecular mechanisms underlying systemic responses remain incompletely understood. Circulating extracellular vesicles (EVs) have emerged as key mediators of interorgan communication, carrying protein cargo that may reflect and influence tissue adaptation to exercise. This study aimed to characterize the proteomic profile of plasma-derived EVs (ExerVs) from exercised versus sedentary mice to uncover candidate proteins involved in exercise-induced remodeling and metabolic regulation. EVs were isolated from plasma of mice subjected to four weeks of voluntary wheel running, followed by untargeted LC-MS/MS analysis using data-dependent acquisition-PASEF on a timsTOF Pro 2. Label-free quantification was performed to identify differentially expressed proteins. These findings provide insight into the EV-mediated mechanisms of exercise adaptation and support future translational efforts to identify systemic exercise-responsive biomarkers and therapeutic targets.