Inhibition of protein and aminoacyl-tRNA synthesis, and binding and transport sites for aromatic amino acids in the brain in vitro with aromatic acids.

The influx of [3H]phenylalanine, [3H]tyrosine and [3H]tryptophan into brain slices and synaptosomes, their binding to synaptic membranes and their incorporation into protein and aminoacyl-tRNA were studied in the presence of an excess of a second aromatic amino acid or some other aromatic acid, viz., phenylpyruvate, phenyllactate, phenylacetate, homogentisate, salicylate or benzoate. The influx into brain slices was strongly inhibited by a second aromatic amino acid and in general also by phenylpyruvate and homogentisate, but the effects of these substances upon the influx into synaptosomes were slight. The binding of phenylalanine and tyrosine to the synaptic membranes was affected mainly by phenylpyruvate and homogentisate, and these were also effective in preventing the formation of aminoacyl-tRNA, and thus apparently inhibited the biosynthesis of proteins and polyphenylalanine. In all cases phenyllactate, phenylacetate salicylate and benzoate had virtually no effect. Phenylalanine seemed to be a noncompetitive, and tyrosine a competitive inhibitor, while tryptophan had both properties, as was also the case with phenylpyruvate and homogentisate. Under phenylketonuric conditions high excesses of phenylalanine and phenylpyruvate, and also certain other aromatic compounds, seemed to occupy the cellular transport sites for amino acids on the cellular membranes and prevent the formation of aminoacyl-tRNAs, thus inhibiting brain protein synthesis. The reduced supply of intracellular amino acids and the inhibition of protein synthesis may constitute one reason for the development of biochemical phenylketonuric abnormalities.