Abstract: | After demonstration that emetine is amebicidal by inhibiting protein synthesis, the question arose whether active protein synthesis is required for emetine's amebicidal effect. The answer appears to be “no,” as derived from experiments on intact amebae. Responses were compared for log- and stationary-growth phase amebae. In the latter, protein synthesis is significantly slower, and sensitivity to emetine, i.e. degree of inhibition of protein synthesis, was maintained independently of rate of protein synthesis. Both stages equally bound tritiated emetine to their ribcsomes. Binding of 3H]emetine was not affected by certain drugs that interfere with energy metabolism, protein synthesis, and/or ribosomal function, e.g. dinitrophenol, puromycin, chloroquine, and acriflavin. High concentrations of EDTA combined with puromycin (which disaggregates ribosomes into their subunits) lowered binding by 50%. In chase experiments the ribosomes of intact amebae were prelabeled with 3H]emetine or 3H]isoemetine, then exposed to relatively high concentrations of unlabeled emetine. Labeled isoemetine was displaced almost completely, whereas no displacement of 3H]emetine occurred; evidently, the high stability of the emetine-ribosome binding is due in part to a hydrogen-bonding reaction of the C-1' atom of the emetine molecule with the chain-elongation site. Finally, evidence was obtained that capacity to bind emetine is an index of drug resistance. |