The effects of processing and cooking practices on the chemical composition and antioxidant activity of Portuguese wild edible mushroom species (Lactarius deliciosus, Macrolepiota mastoidea, Macrolepiota procera, and Sarcodon imbricatus) were investigated. Dried, frozen, and cooked samples were analyzed for proximate constituents (moisture, fat, crude protein, ash, and carbohydrates) and nutritional value. Fatty acid and sugar profiles were also obtained by gas-liquid chromatography/flame ionization detection and high-performance liquid chromatography/refraction index, respectively. The antioxidant properties were evaluated by several biochemical assays: 2,2-diphenyl-1-picrylhydrazyl radical scavenging capacity, reducing power, inhibition of beta-carotene bleaching, and inhibition of lipid peroxidation in brain tissue using thiobarbituric acid reactive substances. Results of this study show that mushroom species and processing and cooking practices are all effective determinants for either chemical composition or antioxidant properties. Cooked samples proved to have lower nutrient concentrations and lower antioxidant activities than either dried or frozen samples. In what concerns fatty acids and sugar individual profiles, only cooking proved to be relevant: The cooked samples presented higher monounsaturated fatty acid and lower polyunsaturated fatty acid and sugars contents.