While ethanol can be metabolized as a nutrient, a downside of its ingestion is thought to be the inhibition of protein synthesis. Current methods used to measure protein synthesis are time-consuming, expensive, and potentially hazardous. Reported here is a simpler method based on light emission by the enzyme luciferase. Human h293 kidney epithelial cells were transfected with a tetracycline inducible luciferase plasmid and the tet activator plasmid. Cell extracts were prepared, and the luciferase activity was determined by the measurement of light emission upon addition of luciferin. To determine the efficacy of this procedure, transfected cells were treated with the well-known protein synthesis inhibitor cycloheximide. Light emission decreased, consistent with inhibition of protein synthesis. Transfected cells were treated with various concentrations of ethanol, which also decreased light emission. Control experiments indicated that, at the levels of ethanol used, the ethanol was not inhibiting the luciferase enzyme directly nor killing the cells. Thus it was concluded that ethanol inhibits the synthesis of protein in these cells. Furthermore, these results suggest that a simplified protein synthesis assay based on luciferase activity can be used to evaluate potential protein synthesis inhibitors like ethanol.

Ethanol inhibition of protein synthesis in mammalian cells

255th National Meeting (2018)

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