Measurement of Protein Synthetic Activity by Determination of Peptidyl[3H]puromycin Formation in Liver Slices after Ethanol Administration

Terrence Donohue, J. H. Sorrell, Michael Floyd Sorrell, D. J. Tuma

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

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 mw 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.

Original languageEnglish (US)
Pages (from-to)526-530
Number of pages5
JournalAlcoholism: Clinical and Experimental Research
Volume9
Issue number6
DOIs
StatePublished - Nov 1985

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Puromycin
Liver
Ethanol
Valine
Rats
Proteins
Nutrition
Reproducibility of Results
peptidyl-puromycin
Animals
Diet
Amino Acids
Peptides
Liquids

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Toxicology
  • Psychiatry and Mental health

Cite this

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title = "Measurement of Protein Synthetic Activity by Determination of Peptidyl[3H]puromycin Formation in Liver Slices after Ethanol Administration",
abstract = "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 mw 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.",
author = "Terrence Donohue and Sorrell, {J. H.} and Sorrell, {Michael Floyd} and Tuma, {D. J.}",
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AU - Donohue, Terrence

AU - Sorrell, J. H.

AU - Sorrell, Michael Floyd

AU - Tuma, D. J.

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N2 - 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 mw 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.

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