The human erythrocyte sugar transporter presents two sugar import sites.
The human erythrocyte sugar transporter presents sugar import (e2) and sugar export (e1) sites simultaneously. This study asks whether the sugar transporter exposes only one or multiple import sites. We approached this question by analysis of cytochalasin B binding to the human erythrocyte sugar export site in the presence of sugars that bind to the sugar import site. Extracellular maltose does not enter human erythrocytes. High concentrations of maltose (1-100 mM) inhibit cytochalasin B binding to human red cells. Low concentrations (25-500 microM) increase the level of erythrocyte cytochalasin B binding. Maltose modulation of cytochalasin B binding is mediated by altered affinity of sugar export sites for cytochalasin B. Similar results are obtained with other cell-impermeant inhibitors of sugar uptake. Extracellular D-glucose (a transported sugar) stimulates cytochalasin B binding at low D-glucose concentrations (10-250 microM), but this effect is lost at higher concentrations. Intracellular D-glucose inhibits cytochalasin B binding. Low concentrations of extracellular maltose and other nontransported inhibitors stimulate 3-O-methylglucose uptake in erythrocytes. Higher sugar concentrations (1-100 mM) inhibit transport. These data support the hypothesis that the erythrocyte sugar transporter presents two sugar import sites and at least one sugar export site. This conclusion is consistent with the proposed oligomeric structure of the sugar transporter, a complex of four GluT1 proteins in which each subunit presents a translocation pathway.
Hamill
S
S
Department of Biochemistry and Molecular Biology, University of Massachusetts Medical School, Worcester 01655, USA.
Cloherty
E K
EK
Carruthers
A
A
D019325:3-O-Methylglucose / Q000097:blood; D001665:Binding Sites / Q000187:drug effects; D002241:Carbohydrates / Q000097:blood* / Q000494:pharmacology; D003571:Cytochalasin B / Q000097:blood; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology; D051272:Glucose Transporter Type 1; D006801:Humans; D008320:Maltose / Q000494:pharmacology; D008954:Models, Biological; D009004:Monosaccharide Transport Proteins / Q000097:blood* / Q000737:chemistry*; D014312:Trisaccharides / Q000494:pharmacology
D002241:Carbohydrates; D051272:Glucose Transporter Type 1; D009004:Monosaccharide Transport Proteins; C495677:SLC2A1 protein, human; D014312:Trisaccharides; D019325:3-O-Methylglucose; D003571:Cytochalasin B; C008317:maltotriose; D008320:Maltose; D005947:Glucose