Measurement of protein synthetic activity by determination of peptidyl[3H]puromycin formation in liver slices after ethanol administration.
We investigated the utility of [3H]puromycin as an alternate and adjunct precursor to amino acids for measuring protein synthetic activity in rat liver slices. Slices were incubated in the presence of either [3H]puromycin or radiolabeled valine to compare the incorporation of these isotopic precursors into nascent hepatocellular proteins. Compared to liver slices from controls, comparable decreases in the incorporation of both [3H]puromycin and labeled valine were observed in experiments using slices from fasted rats and in slices preincubated with 25 mM ethanol. Radiolabeling of nascent polypeptides with either [3H]puromycin or labeled valine in liver slices from rats fed a liquid diet containing ethanol was also decreased compared to slices from pair-fed control and chow-fed animals. Our results demonstrated the validity of using [3H]puromycin to detect changes in protein synthetic activity under these conditions. The potential advantage of using [3H]puromycin for in vivo studies is discussed.
D000437:Alcoholism / Q000378:metabolism; D000818:Animals; D046508:Culture Techniques; D000431:Ethanol / Q000494:pharmacology*; D007700:Kinetics; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D011506:Proteins / Q000378:metabolism*; D011691:Puromycin / Q000031:analogs & derivatives* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D014633:Valine / Q000378:metabolism
D011506:Proteins; C031736:peptidyl-puromycin; D000431:Ethanol; D011691:Puromycin; D014633:Valine