Increasing the plasma glucose and insulin concentrations during prolonged variable intensity exercise by supplementing with carbohydrate has been found to spare muscle glycogen and increase aerobic endurance. Furthermore, the addition of protein to a carbohydrate supplement will enhance the insulin response of a carbohydrate supplement. The purpose of the present study was to compare the effects of a carbohydrate and a carbohydrate-protein supplement on aerobic endurance performance. Nine trained cyclists exercised on 3 separate occasions at intensities that varied between 45% and 75% VO2max for 3 h and then at 85% VO2max until fatigued. Supplements (200 ml) were provided every 20 min and consisted of placebo, a 7.75% carbohydrate solution, and a 7.75% carbohydrate/1.94% protein solution. Treatments were administered using a double-blind randomized design. Carbohydrate supplementation significantly increased time to exhaustion (carbohydrate 19.7 +/- 4.6 min vs. placebo 12.7 +/- 3.1 min), while the addition of protein enhanced the effect of the carbohydrate supplement (carbohydrate-protein 26.9 +/- 4.5 min, p < .05). Blood glucose and plasma insulin levels were elevated above placebo during carbohydrate and carbohydrate-protein supplementation, but no differences were found between the carbohydrate and carbohydrate-protein treatments. In summary, we found that the addition of protein to a carbohydrate supplement enhanced aerobic endurance performance above that which occurred with carbohydrate alone, but the reason for this improvement in performance was not evident.