Energy deficit and dietary macronutrient intake are thought to independently modulate cognition, mood and sleep. To what extent manipulating the dietary ratio of protein-to-carbohydrate affects mood, cognition and sleep during short-term energy deficit is undetermined. Using a randomized, block design, 39 non-obese young adults (21 ± 1 years, BMI 25 ± 1 kg/m2) consumed diets containing 0.8 g, 1.6 g or 2.4 g protein per kg body weight per day for 31 days. Carbohydrate intake was reduced to accommodate higher protein intakes while dietary fat was maintained at 30% of total energy intake. Cognitive performance, mood, self-reported sleep quality, and plasma amino acid concentrations were periodically assessed during a 10-day energy balance period and a subsequent 21 -day, 40% energy deficit period. Anger, tension and total mood disturbance increased during the initial ten days of energy deficit (P < 0.05), but by the end of the energy deficit returned to levels not different from those measured during energy balance. No effects of dietary protein-to-carbohydrate ratio on cognitive performance, mood or self-reported sleep quality were observed during energy balance or energy deficit. Thus, high-protein, low-carbohydrate, moderate-fat diets do not appear to benefit or impair cognition, mood or sleep in non-obese adults during energy deficit. These findings suggest that energy deficit may initially be psychologically difficult for non-obese individuals attempting to lose weight, but that these changes are transient. Employing strategies that alleviate decrements in mood during this initial period of adaptation may help sustain weight loss efforts.