Protection of alveolar macrophages from hyperoxia by gamma-glutamyl transpeptidase.
Exposure to hyperoxia causes loss of alveolar macrophage cell function. Toxicity was measured as suppression of the respiratory burst stimulated by phorbol myristate acetate subsequent to exposure (43.5% depression by 2-h exposure to 5 atm absolute O2 vs. controls). The presence of extracellular glutathione significantly protected these cells (7% loss). gamma-Glutamyl transpeptidase, a membrane enzyme with its active site directed outward, was necessary for use of extracellular glutathione. This was demonstrated using the gamma-glutamyl transpeptidase inhibitor, serine-borate complex, which significantly blocked both protection of cells by extracellular glutathione and extracellular glutathione-dependent synthesis of glutathione. The principal use of glutathione in antioxidant defense is as a substrate for glutathione peroxidase. The apparent Km for glutathione of glutathione peroxidase of rat alveolar macrophages was determined to be 2 mM; however, rat alveolar macrophages have approximately 1.3 mM intracellular glutathione, which is insufficient for maximal enzymatic activity. During hyperoxic exposure, this deficit would probably be more significant. Thus the ability of extracellular glutathione along with gamma-glutamyl transpeptidase activity to provide amino acids for de novo glutathione synthesis appears to be a potentially important component of antioxidant defense.
Forman
H J
HJ
Department of Pediatrics, University of Southern California, Childrens Hospital, Los Angeles 90027.
Skelton
D C
DC
D000818:Animals; D002478:Cells, Cultured; D005978:Glutathione / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000378:metabolism; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D008298:Maleates / Q000494:pharmacology; D010100:Oxygen / Q000494:pharmacology* / Q000633:toxicity; D051381:Rats; D011919:Rats, Inbred Strains; D013481:Superoxides / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D005723:gamma-Glutamyltransferase / Q000378:metabolism*
D008298:Maleates; D013481:Superoxides; D007770:L-Lactate Dehydrogenase; D005979:Glutathione Peroxidase; D005723:gamma-Glutamyltransferase; C014476:diethyl maleate; D005978:Glutathione; D013755:Tetradecanoylphorbol Acetate; D010100:Oxygen