The glucose transporter in the plasma membrane of the outer segments of bovine retinal rods.
The facilitated diffusion glucose transporter in the plasma membrane of intact outer segments isolated from bovine retinal rods (ROS) was characterized by measurements of: (1) 14C-labeled 3-O-methylglucose fluxes; and (2) glucose-sensitive binding of 3H-labeled cytochalasin B to ROS membranes. Inhibition of 3-O-methylglucose influx into ROS, inhibition of 3-O-methylglucose efflux from ROS and glucose-sensitive binding of cytochalasin B to ROS showed very similar cytochalasin B inhibition/dissociation constants of 0.9 microM, 1.3 microM and 1.3 microM, respectively. D-glucose inhibited both 14C-labeled 3-O-methylglucose transport and cytochalasin B binding. The above results suggest that D-glucose-sensitive cytochalasin B binding reflects specific binding to the ROS glucose transporter and the density of glucose transporter in the ROS plasma membrane was determined to be 800 microns-2, comparable to relatively abundant ROS plasma membrane proteins such as the cGMP-gated channel and the Na-Ca+K exchanger. Displacement of 3H-labeled cytochalasin B by non-transportable hexoses was used to localize the hexose transporters to the ROS plasma membrane and to examine a simple single-site, alternating conformation model for hexose transport. A comparison between the Glut1 glucose transporters of bovine ROS, bovine erythrocytes and human erythrocytes suggests that kinetic and pharmacological characteristics of glucose transporters cannot be predicted in a simple manner from gene type and species.
Li
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XB
Department of Medical Biochemistry, University of Calgary, Alberta, Canada.
Szerencsei
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RT
Schnetkamp
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PP
D019325:3-O-Methylglucose; D000818:Animals; D001665:Binding Sites; D001786:Blood Glucose / Q000378:metabolism; D002417:Cattle; D003571:Cytochalasin B / Q000494:pharmacology; D004912:Erythrocytes / Q000378:metabolism; D066298:In Vitro Techniques; D008757:Methylglucosides / Q000378:metabolism; D009004:Monosaccharide Transport Proteins / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000378:metabolism*
D001786:Blood Glucose; D008757:Methylglucosides; D009004:Monosaccharide Transport Proteins; D019325:3-O-Methylglucose; D003571:Cytochalasin B