The distribution of the four macronutrients is associated with energy intake and body fatness according to short-term interventions. The present study involves macronutrient distribution in relation to energy intake and body fatness over a period of 23 years in individuals who have ad libitum access to food. Eight follow-up measurements have been performed in 168 men and 182 women who participate in the Amsterdam Growth and Health Longitudinal Study. From the age of 13 years onwards, dietary intake, physical activity and the thickness of four skinfolds have been assessed. Body fatness was assessed using dual-energy X-ray absorptiometry at the age of 36 years. Generalised estimating equation regression analyses showed that energy percentages (En%) from protein and (in men) carbohydrates were inversely related to energy intake, while the En% from fat was positively related with energy intake. The men and women with high body fatness at the age of 36 years had a 1 En% higher protein intake, and the women with high body fatness had a 2 En% lower alcohol intake at the age of 32 and 36 years. The apparent inconsistent relationships between protein and energy intake and protein and body fatness can in women be explained by reverse causation and underreporting, as in women, low energy intake could not be explained by low physical activity. In conclusion, high intake of protein and (in men) carbohydrate, and low intake of fat are inversely related to total energy intake. High body fatness at the age of 36 years is related to a higher protein intake and, in women, to a lower alcohol intake.