Protein synthesis in skeletal muscle is known to decrease during exercise and it has been suggested that this may depend on the magnitude of the relative metabolic stress within the contracting muscle. To examine the mechanisms behind this, the effect of exercise intensity on skeletal muscle eukaryotic elongation factor 2 (eEF2) and eukaryotic initiation factor 4E binding protein 1 (4EBP1) phosphorylation, key components in the mRNA translation machinery, were examined together with AMP activated protein kinase (AMPK) in healthy young men. Skeletal muscle eEF2 phosphorylation at Thr⁵⁶ increased during exercise but was not influenced by exercise intensity, and was lower than rest 30min after exercise. On the other hand, 4EBP1 phosphorylation at Thr^37/46 decreased during exercise and this decrease was greater at higher exercise intensities, and was similar to rest 30min after exercise. AMPK activity, as indexed by AMPK -subunit phosphorylation at Thr¹⁷² and phosphorylation of the AMPK substrate ACC at Ser²²¹, was higher with higher exercise intensities, and these indices were higher than rest after high intensity exercise only. Using immunohistochemistry, it was shown that the increase in skeletal muscle eEF2 Thr⁵⁶ phosphorylation was restricted to type I myofibers. Taken together, these data suggest that the depression of skeletal muscle protein synthesis with endurance-type exercise may be regulated at both initiation (i.e. 4EBP1) and elongation (i.e. eEF2) steps, with eEF2 phosphorylation contributing at all exercise intensities but 4EBP1 dephosphorylation contributing to a greater extent at high versus low exercise intensities.