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J. Biol. Chem., Vol. 276, Issue 16, 12725-12729, April 20, 2001
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From the Departments of The transport of glucose across the mammalian
blood-brain barrier is mediated by the GLUT1 glucose transporter, which
is concentrated in the endothelial cells of the cerebral microvessels.
Several studies supported an asymmetric distribution of GLUT1 protein between the luminal and abluminal membranes (1:4) with a significant proportion of intracellular transporters. In this study we investigated the activity and concentration of GLUT1 in isolated luminal and abluminal membrane fractions of bovine brain endothelial cells. Glucose
transport activity and glucose transporter concentration, as determined
by cytochalasin B binding, were 2-fold greater in the luminal than in
the abluminal membranes. In contrast, Western blot analysis using a
rabbit polyclonal antibody raised against the C-terminal 20 amino acids
of GLUT1 indicated a 1:5 luminal:abluminal distribution. Western blot
analysis with antibodies raised against either the intracellular loop
of GLUT1 or the purified erythrocyte protein exhibited
luminal:abluminal ratios of 1:1. A similar ratio was observed when the
luminal and abluminal fractions were exposed to the
2-N-4[3H](1-azi-2,2,2,-trifluoroethyl)benzoxyl-1,3-bis-(d-mannos-4-yloxyl)-2-propylamine ([3H]ATB-BMPA) photoaffinity label. These
observations suggest that either an additional glucose transporter
isoform is present in the luminal membrane of the bovine blood-brain
barrier or the C-terminal epitope of GLUT1 is "masked" in the
luminal membrane but not in the abluminal membranes.
Glucose Transporter Asymmetries in the Bovine Blood-Brain
Barrier*
§,
, and
Neuroscience and Anatomy and
¶ Pediatrics, Milton S. Hershey Medical Center, Pennsylvania State
University College of Medicine, Hershey, Pennsylvania 17033 and the
Department of Physiology and Biophysics, Finch University of
Health Science, The Chicago Medical School,
North Chicago, Illinois 60064-3095
*
This work was supported by NINDS National Institutes of
Health Grant NS 31017 (to R. A. H.).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.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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