Inhibition of aminoacyl-tRNA synthetases by p-chloroamphetamine and its role in protein synthesis inhibition.
p-Chloroamphetamine inhibited to some degree all amino acid-dependent pyrophosphate-exchange activities which could be detected in a rabbit reticulocyte extract. A detailed kinetic analysis of the reaction catalyzed by reticulocyte leucyl-tRNA synthetase demonstrated that the inhibitor affected only amino acid binding. Less rigorous studies of other synthetases from both rabbit reticulocyte and Escherichia coli could be similarly interpreted, suggesting that this compound interacts in a common manner with these several enzymes. The contribution of such effects to the inhibition of protein synthesis by the drug was investigated using cell-free translation systems in which rates of amino acid incorporation were limited to varying degrees by the synthesis and availability of aminoacyl-tRNA. In a wheat germ system programmed with globin mRNA, in which levels of amino acids and aminoacyl-tRNAs were shown to limit the rate of protein synthesis, the inhibition produced by p-chloroamphetamine could be partially reversed by increasing the concentration of the limiting amino acid. In a reticulocyte lysate, in which amino acid concentrations were not limiting, inhibition failed to show an amino acid-reversible component. Thus, while the inhibition of aminoacyl-tRNA synthetases by amphetamines can be shown in some cases to play a role in the effects of these compounds on in vitro protein synthesis, other sites of interference with initiation and/or elongation reactions may predominate, depending on the construction of the system under study.
D000604:Amino Acyl-tRNA Synthetases / Q000037:antagonists & inhibitors*; D000662:Amphetamines / Q000494:pharmacology*; D000818:Animals; D007700:Kinetics; D007935:Leucine-tRNA Ligase / Q000037:antagonists & inhibitors*; D008274:Magnesium / Q000494:pharmacology; D011072:Poly U / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects*; D011817:Rabbits; D012156:Reticulocytes / Q000187:drug effects / Q000378:metabolism*; D012270:Ribosomes / Q000187:drug effects / Q000378:metabolism*; D012639:Seeds / Q000187:drug effects / Q000378:metabolism; D014908:Triticum / Q000378:metabolism; D010133:p-Chloroamphetamine / Q000494:pharmacology*
D000662:Amphetamines; D011072:Poly U; D010133:p-Chloroamphetamine; D000604:Amino Acyl-tRNA Synthetases; D007935:Leucine-tRNA Ligase; D008274:Magnesium