The potential relationship between physical activity and endogenous pain modulatory capacity remains unclear. Therefore, the aim of the current study was to compare the pain modulatory responses of athletes and non-athletes. Conditioned pain modulation (CPM) was assessed in 15 athletes and 15 non-athletes at rest. Participation was restricted to pain-free males between 18 and 40 years of age. To measure CPM capacity, a sequential CPM testing protocol was implemented, whereby a test stimulus (pressure pain threshold [PPT]) was presented before and immediately after a conditioning stimulus (4-min cold-pressor test). Pain intensity ratings were obtained at 15-s intervals throughout the cold-pressor task using a numerical rating scale. Athletes demonstrated higher baseline PPTs compared to non-athletes (P = .03). Athletes also gave lower mean (P < .001) and maximum (P < .001) pain intensity ratings in response to the conditioning stimulus. The conditioning stimulus had a stronger inhibitory effect on the test stimulus in athletes, showing enhanced CPM in athletes compared to non-athletes (P < .05). This finding of enhanced CPM in athletes helps clarify previous mixed findings. Potential implications for exercise performance and injury are discussed.