The involvement of sulphydryl groups in the peptidyl transferase centre of eukaryotic ribosomes.
Treatment of mammalian ribosomes with N-ethylmaleimide enhances up to 100% the ribosome efficiency in the "fragment reaction assay" for peptide bond formation by increasing the affinity of the substrate C-A-C-C-A-Leu-Ac for the donor site. This stimulation in peptidyl transferase activity was not observed when yeast ribosomes were treated in a similar manner. Stimulation of the peptidyl transferase activity of mammalian ribosomes was also observed by treatment with either p-chloromercuribenzoic acid or 5,5'-dithiobis-(2-nitrobenzoic acid) or 5,5'-dithiobis-(2-nitropyridine) or the maleimide-derived antibiotic showdomycin. N-Ethylmaleimide treatment also enhances C-A-C-C-A-Leu binding to the acceptor site of the peptidyl transferase centre. However, neither binding of N-Ac-Phe-tRNA in the presence of ethanol, nor binding of Phe-tRNA to the ribosomes is stimulated by N-ethylmaleimide. The antibiotic tenuazionic acid (a selective inhibitor of peptide bond formation by mammalian ribosomes) appears to require for its inhibitory effect the ribosome sulphydryl residues, since its inhibitory action on the fragment reaction is greatly decreased in ribosomes treated with N-ethylmaleimide.
D000217:Acyltransferases / Q000378:metabolism*; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001565:Benzoates / Q000494:pharmacology; D002729:Chloromercuribenzoates / Q000494:pharmacology; D000431:Ethanol / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D007700:Kinetics; D009574:Nitro Compounds / Q000494:pharmacology; D011691:Puromycin / Q000494:pharmacology; D011817:Rabbits; D012156:Reticulocytes / Q000201:enzymology; D012270:Ribosomes / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D013045:Species Specificity; D013438:Sulfhydryl Compounds / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D001565:Benzoates; D002729:Chloromercuribenzoates; D009574:Nitro Compounds; D013438:Sulfhydryl Compounds; D000431:Ethanol; D011691:Puromycin; D000217:Acyltransferases; D005033:Ethylmaleimide