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Measurement of Protein Synthetic Activity by Determination of Peptidyl[ 3 H]puromycin Formation in Liver Slices after Ethanol Administration
Author(s) -
Donohue Terrence M.,
Sorrell James H.,
Sorrell Michael F.,
Tuma Dean J.
Publication year - 1985
Publication title -
alcoholism: clinical and experimental research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.267
H-Index - 153
eISSN - 1530-0277
pISSN - 0145-6008
DOI - 10.1111/j.1530-0277.1985.tb05597.x
Subject(s) - puromycin , valine , in vivo , ethanol , chemistry , amino acid , in vitro , protein biosynthesis , biochemistry , biology , microbiology and biotechnology
We investigated the utility of [ 3 H]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 [ 3 H]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 [ 3 H]puromycin and labeled valine were observed in experiments using slices from fasted rats and in slices preincubated with 25 mw ethanol. Radiolabeling of nascent polypeptides with either [ 3 H]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 [ 3 H]puromycin to detect changes in protein synthetic activity under these conditions. The potential advantage of using [ 3 H]puromycin for in vivo studies is discussed.