Amino acid inhibition of the autophagic/lysosomal pathway of protein degradation in isolated rat hepatocytes.
Protein degradation in isolated rat hepatocytes, as measured by the release of [14C]valine from pre-labelled protein, is partly inhibited by a physiologically balanced mixture of amino acids. The inhibition is largely due to the seven amino acids leucine, phenylalanine, tyrosine, tryptophan, histidine, asparagine and glutamine. When the amino acids are tested individually at different concentrations, asparagine and glutamine are the strongest inhibitors. However, when various combinations are tested, a mixture of the first five amino acids as well as a combination of leucine and asparagine inhibit protein degradation particularly strongly. The inhibition brought about by asparagine plus leucine is not additive to the inhibition by propylamine, a lysosomotropic inhibitor; thus indicating that the amino acids act exclusively upon the lysosomal pathway of protein degradation. Following a lag of about 15 min the effect of asparagine plus leucine is maximal and equal to the effect of propylamine, suggesting that their inhibition of the lysosomal pathway is complete as well as specific. Degradation of endocytosed 125I-labelled asialofetuin is not affected by asparagine plus leucine, indicating that the amino acids do not affect lysosomes directly, but rather inhibit autophagy at a step prior to the fusion of autophagic vacuoles with lysosomes. The aminotransferase inhibitor, aminooxyacetate, does not prevent the inhibitory effect of any of the amino acids, i.e. amino acid metabolites are apparently not involved.
D000596:Amino Acids / Q000494:pharmacology*; D000625:Aminooxyacetic Acid / Q000494:pharmacology; D000818:Animals; D001216:Asparagine / Q000494:pharmacology; D001343:Autophagy / Q000187:drug effects*; D004357:Drug Synergism; D005973:Glutamine / Q000494:pharmacology; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism*; D008297:Male; D010587:Phagocytosis / Q000187:drug effects*; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D000637:Transaminases / Q000037:antagonists & inhibitors
D000596:Amino Acids; D011506:Proteins; D005973:Glutamine; D000625:Aminooxyacetic Acid; D001216:Asparagine; D000637:Transaminases