EFFECTS OF DIHYDROSTREPTOMYCIN ON AMINO ACID INCORPORATION INTO THE PROTEINS OF M. TUBERCULOSIS (BCG)

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Abstract
A study has been made of the effects of dihydrostreptomycin on amino acid incorporation into the proteins of M. tuberculosis (BCG). Suspensions of this organism on incubation at 37[degree] with glycine-l-C14 give rise, aerobically, to labelled proteins in which 80% of the radioactivity appears in the glycine and serine moieties of the proteins and about 20% in alanine and aspartic acid. In presence of glycine-2-C14, radioactivity appears in a larger number of amino acids of the protein. Incubation with serine-3-C14 leads to a distribution of radioactivity in the amino acids in BCG proteins but alanine-1-C14 and valine-l-C14 give rise to proteins with the radioactivity almost entirely in the corresponding amino acids. The process of aerobic incorporation of radioactivity from glycine-1-C14 in BCG proteins is stimulated by the presence of glucose, glycerol, sodium pyruvate, sodium stearate, or sodium benzoate in the medium in which the cells are incubated, the rate of incorporation being approximately constant over a period of 4 hours. The incorporation depends largely on the presence of oxygen. Dihydrostreptomycin (33 [mu]g per ml) markedly inhibits labelling of proteins in the cell suspensions in presence of radioactive amino acids, the inhibition increasing with concentration of the streptomycin to an optimal concentration of 200 [mu]g/ml. Penicillin and isonicotinic hydrazide are inactive but chloromycetin is an effective inhibitor. Cyanide, arsenite, and azide are inhibitory. The presence of lecithin stimulates incorporation of radioactivity from glycine- 1-C14 into BCG proteins. Dihydrostreptomycin inhibitors of amino acid incorporation into BCG proteins increase with time of incubation of the cells with the drug. Concentrations of dihydrostreptomycin that inhibit labelled amino acid incorporation into labelled proteins by 50% have no effect on BCG respiration. The drug has no inhibitory effect on labelled amino acid incorporation in E. coli or Ehrlich ascites carcinoma cells in vitro but is effective with M. phlei. It does not affect selectively the distribution of radioactivities of the component amino acids of BCG proteins; only the total radioactivity incorporated into the proteins is diminished. The results lead to the conclusion that dihydrostreptomycin brings about an inhibition of protein synthesis in the BCG strain of M. tuberculosis at concentrations at which it exerts antibiotic effects.