The effect of some ionophores and metabolic inhibitors on reticulocytes protein synthesis was examined. At microM concentrations, valinomycin, nigericin and CCCP rapidly inhibit protein synthesis, while with antimycin-A or DCCD the inhibition is rather slow. The onset of the arrest of protein synthesis coincides with a 20--30% drop in the intracellular ATP content. No inhibition in protein synthesis or drop in ATP was found after 1 h incubation without glucose or in the presence of 2-deoxyglucose. It is shown that the inhibition by valinomycin, nigericin or CCCP is not due to their effect on K+ and/or H+ fluxes through the plasma membrane. Reticulocytes incubated at pH 8.2 show much lower inhibition of protein synthesis by nigericin, CCCP, DCCD or antimycin-A. On the other hand, at this alkaline pH, starvation to glucose causes high inhibition of protein synthesis. It is concluded that the ionophores inhibit protein synthesis due to their uncoupling effect on mitochondrial ATP synthesis. At high pH, the glycolytic activity is relatively high, and the ATP generated by the glycolysis can compensate to some degree for the ATP loss in the oxidative phosphorylation.