The effects of adaptation to a high-fat diet on endurance performance are equivocal, and there is little data regarding the effects on high-intensity exercise performance. This study examined the effects of a high-fat/moderate protein diet on submaximal, maximal, and supramaximal performance. Twenty non-highly trained men were assigned to either a high-fat/moderate protein (HFMP; 61% fat diet) (n = 12) or a control (C; 25% fat) group (n = 8). A maximal oxygen consumption test, two 30-s Wingate anaerobic tests, and a 45-min timed ride were performed before and after 6 weeks of diet and training. Body mass decreased significantly (-2.2 kg; p < or = .05) in HFMP subjects. Maximal oxygen consumption significantly decreased in the HFMP group (3.5 +/- 0.14 to 3.27 +/- 0.09 L x min(-1)) but was unaffected when corrected for body mass. Perceived exertion was significantly higher during this test in the HFMP group. Main time effects indicated that peak and mean power decreased significantly during bout 1 of the Wingate sprints in the HFMP (-10 and -20%, respectively) group but not the C (-8 and -16%, respectively) group. Only peak power was lower during bout 1 in the HFMP group when corrected for body mass. Despite significantly reduced RER values in the HFMP group during the 45-min cycling bout, work output was significantly decreased (-18%). Adaptation to a 6-week HFMP diet in non-highly trained men resulted in increased fat oxidation during exercise and small decrements in peak power output and endurance performance. These deleterious effects on exercise performance may be accounted for in part by a reduction in body mass and/or increased ratings of perceived exertion.