Objective: Obesity and insulin resistance are associated with muscle mitochondrial dysfunction, which might be related to impairment of mitochondrial protein synthesis. This study aimed at investigating mixed and mitochondrial protein synthesis in skeletal muscle in response to dietary manipulations. Research Methods and Procedures: High-sucrose (SU) and high-fat, high-sucrose (F) diets were provided for 6 weeks to Wistar rats at standard (N) and high (H) energy intakes and compared with controls. Fractional synthesis rates of mixed (FSRPT) and mitochondrial (FSRm) proteins within the oxidative (soleus) and glycolytic (tibialis) muscles were measured using stable isotope flooding dose technique using l-[13C]-valine. Carbonyl content, citrate synthase, and cytochrome c oxidase activities were assayed spectrophotometrically on isolated mitochondria. Results: In the soleus, FSRPT was increased by 40% in the NSU and NF groups and by 65% in the HSU and HF groups (p < 0.001 vs. control). FSRm was increased with high-fat diets (NF, +16%; HF, +32%; p < 0.01). In the tibialis, FSRPT was enhanced in all experimental groups (+31% to 37%, p < 0.05 vs. control). FSRm was augmented in the NSU, NF, and HF groups (+28% to 32%, p < 0.01). Cytochrome c oxidase activity was significantly decreased in all experimental groups in the soleus (p < 0.001). Discussion: Muscle mixed and mitochondrial protein FSR are enhanced after short-term dietary intervention known to induce insulin resistance and obesity. Adaptations are muscle type specific and may not explain alterations in mitochondrial oxidative capacity but might contribute to maintain mitochondrial functioning.