High glutathione turnover in human cell lines revealed by acivicin inhibition of gamma-glutamyltranspeptidase and the effects of thiol-reactive metals during acivicin inhibition

Clin Chim Acta. 2004 Nov;349(1-2):45-52. doi: 10.1016/j.cccn.2004.05.024.


Background: Glutathione is the most abundant nonprotein sulfhydryl-containing compound and constitutes the largest component of the endogenous thiol buffer. Glutathione is known to have multifaceted physiological functions and is a critical factor in protecting organisms against toxicity and disease. Intracellular cysteine concentration is a limiting factor for glutathione synthesis.

Methods: In the present study, the metabolism of intra- and extracellular glutathione in HeLa and hepatoma cell cultures is investigated by using different transport inhibitors for cellular uptake of cystine/cysteine.

Results: There exist several ways of cystine/cysteine transport into HeLa and hepatoma cells, and inhibition of them decreased intracellular concentration of cystine/cysteine and in some cases also of glutathione. It was also shown that a large pool of total cell culture glutathione was located extracellularly in both HeLa and hepatoma cell cultures when gamma-glutamyltranspeptidase (GT) activity was inhibited by acivicin (ACI). Furthermore, the addition of thiol-reactive metal ions significantly increased the total amount of glutathione in hepatoma cell cultures during acivicin inhibition. Thus, occasional determinations of extracellular concentrations of glutathione without GT inhibition strongly underestimate the total turnover of glutathione in a cell culture.

Conclusion: This finding has important implications for future research in glutathione metabolism and the understanding of its role in human health and disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carcinoma, Hepatocellular / metabolism
  • Cell Line, Tumor
  • Cysteine / metabolism
  • Enzyme Inhibitors / pharmacology*
  • Glutathione / metabolism*
  • HeLa Cells
  • Humans
  • Isoxazoles / pharmacology*
  • Liver Neoplasms / metabolism
  • Metals / chemistry*
  • Sulfhydryl Compounds / chemistry*
  • gamma-Glutamyltransferase / antagonists & inhibitors*


  • Enzyme Inhibitors
  • Isoxazoles
  • Metals
  • Sulfhydryl Compounds
  • gamma-Glutamyltransferase
  • Glutathione
  • Cysteine
  • acivicin