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Am J Physiol Cell Physiol (April 9, 2003). doi:10.1152/ajpcell.00060.2003
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Submitted on February 12, 2003
Accepted on April 4, 2003

Distribution of Glut1 in Detergent-Resistant Membranes (DRMs) and non-DRM domains: Effect of Treatment with Azide

Darrell Rubin1 and Faramarz Ismail-Beigi2*

1 Department of Pathology, Case Western Reserve University, Cleveland, OH, USA
2 Department of Medicine and Physiology and Biophysics, Case Western Reserve University, Cleveland, OH, USA

* To whom correspondence should be addressed. E-mail: fxi2{at}po.cwru.edu.

We have previously shown that the acute stimulation of glucose transport in Clone 9 cells in response to azide is mediated by activation of Glut1 and that stomatin, a Glut1-binding protein, appears to inhibit Glut1 function. In Clone 9 cells under basal conditions, ~38% of Glut1, ~70% of stomatin, and the bulk of caveolin-1, was localized in the detergent-resistant membrane (DRM) fraction; a significant fraction of Glut1 is also present in DRMs of 3T3-L1 fibroblasts and human RBCs. Acute exposure to azide resulted in 40 and 50% decreases in the content of Glut1 in DRMs of Clone 9 cells and 3T3-L1 fibroblasts respectively, while the distribution of stomatin and caveolin-1 in Clone 9 cells remained unchanged. In addition, treatment of Clone 9 cells with azide resulted in a ~50% decrease in the content of Glut1 in the DRM fraction of plasma membranes. We conclude that 1) a significant fraction of Glut1 is localized in DRMs, and 2) treatment of cells with azide results in a partial redistribution of Glut1 out of the DRM fraction.




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