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M111053200v1
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Papers In Press, published online ahead of print March 19, 2002
J. Biol. Chem, 10.1074/jbc.M111053200
Submitted on November 19, 2001
Revised on March 5, 2002
Accepted on March 19, 2002

Diphenyleneiodonium triggers the efflux of glutathione from cultured cells

Juliet M. Pullar and Mark B. Hampton

Pathology, Christchurch School of Medicine, Christchurch

Corresponding Author: juliet.pullar{at}chmeds.ac.nz

Diphenyleneiodonium (DPI) is a broad-spectrum flavoprotein inhibitor commonly used to inhibit oxidant production by the NADPH oxidase of phagocytic and non-phagocytic cells. A previous study has shown that DPI can sensitize T24 bladder carcinoma cells to Fas-mediated apoptosis. We observed DPI to deplete intracellular reduced glutathione (GSH) in T24 cells, and a range of other primary and transformed cell types. The effect was immediate, with 50% loss of intracellular GSH within two hours of treatment with DPI. The glutathione was quantitatively recovered in the extracellular medium, indicating that efflux was occurring. The loss of GSH was blocked with bromosulfophthalein, an inhibitor of the canalicular GSH transporters. We conclude that DPI induces a dramatic efflux of cellular GSH from T24 cells via a specific transport channel. This provides a potential mechanism for its pro-apoptotic effect, and it also has important implications for the regulation of glutathione homeostasis in cells.


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