J. G. Pace, M. R. Watts and W. J. Canterbury. T-2 mycotoxin inhibits mitochondrial protein synthesis. Toxicon 26, 77 – 85 1988. — We investigated the effect of T-2 toxin on rat liver mitochondrial protein synthesis. Isolated rat liver mitochondria were supplemented with an S-100 supernatant from rat liver and an external ATP-generating system. We used an in vitro assay employing cycloheximide, an inhibitor of cytoplasmic protein synthesis, and chloramphenicol, an inhibitor of mitochondrial protein synthesis, to distinguish mitochondrial protein synthesis from the cytoplasmic process. Amino acid incorporation into mitochondria was dependent on the concentration of mitochondria and was inhibited by chloramphenicol. The rate of uptake of [ 3H]leucine into mitochondrial protein was unaffected by the addition of T-2 toxin and was not a rate-limiting step in incorporation. However, 0.02 μg/ml of T-2 toxin decreased the rate of protein synthesis by isolated mitochondria by 50%. The degree of protein synthesis inhibition correlated with the amount of T-2 toxin taken up by the mitochondria. While T-2 toxin is known to inhibit eukaryotic protein synthesis, this is the first time T-2 was shown to inhibit mitochondrial protein synthesis.