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Originally published In Press as doi:10.1074/jbc.M111053200 on March 19, 2002

J. Biol. Chem., Vol. 277, Issue 22, 19402-19407, May 31, 2002
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Diphenyleneiodonium Triggers the Efflux of Glutathione from Cultured Cells*

Juliet M. PullarDagger and Mark B. Hampton§

From the Free Radical Research Group, Department of Pathology, Christchurch School of Medicine and Health Sciences, P. O. Box 4345, Christchurch, New Zealand

Diphenyleneiodonium (DPI) is a broad-spectrum flavoprotein inhibitor commonly used to inhibit oxidant production by the NADPH oxidase of phagocytic and nonphagocytic 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 2 h 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 proapoptotic effect, and it also has important implications for the regulation of glutathione homeostasis in cells.


* This work was supported in part by an Otago University Research Grant and by the Cancer Society of New Zealand.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed. Tel.: 64-3-364-0565; Fax: 64-3-364-1083; E-mail: juliet.pullar@chmeds.ac.nz.

§ Supported by a grant from the Marsden Fund of the Royal Society of New Zealand.


Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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