Characterization of GLUT3 protein expressed in Chinese hamster ovary cells
- 15 November 1992
- journal article
- Published by Portland Press Ltd. in Biochemical Journal
- Vol. 288 (1), 189-193
- https://doi.org/10.1042/bj2880189
Abstract
We have expressed GLUT3 protein, an isoform of a facilitative glucose transporter, in Chinese hamster ovary cells by transfection of its cDNA using an expression vector. The expressed GLUT3 protein was detected by Western-blot analysis as a broad band of 45-65 kDa, indicating intensive glycosylation of the protein. The expressed GLUT3 protein was observed, by immunofluorescence staining, to be located mainly at the plasma membrane, and its expression was associated with a marked increase in glucose-transport activity. Kinetic analysis revealed that the Km value of GLUT3 protein for 3-O-methylglucose uptake was approx. 35% of that of GLUT1 protein, whereas the Km value of GLUT3 protein for 2-deoxy-D-glucose uptake was very similar to that of GLUT1 protein. The Vmax. value of GLUT3 protein for 3-O-methylglucose and 2-deoxyglucose uptake was approx. 20-50% of that of GLUT1 protein. GLUT3 protein was well photolabelled with [3H]cytochalasin B or a mannose derivative, 2-N-4-[3H](1-azi-2,2,2-trifluoroethyl)benzoyl-1,3-bis-(D-mannos -4-yloxy)-2- propylamine. Thus GLUT3 protein has very similar characteristics to GLUT1 protein including its subcellular localization, but exhibits lower Km and Vmax. values for 3-O-methylglucose uptake.Keywords
This publication has 27 references indexed in Scilit:
- Intracellular targeting of the insulin-regulatable glucose transporter (GLUT4) is isoform specific and independent of cell type.The Journal of cell biology, 1991
- Tissue distribution and species difference of the brain type glucose transporter (GLUT3)Biochemical and Biophysical Research Communications, 1991
- Erythrocyte/HEPG2-type glucose transporter is concentrated in cells of blood-tissue barriersBiochemical and Biophysical Research Communications, 1990
- C-terminal truncated glucose transporter is locked into an inward-facing form without transport activityNature, 1990
- Regulation of Glucose-Transporter FunctionDiabetes Care, 1990
- Cloning of a rabbit brain glucose transporter cDNA and alteration of glucose transporter mRNA during tissue developmentBiochemical and Biophysical Research Communications, 1988
- Hexose transport in L6 rat myoblasts. I. Rate‐limiting step, kinetic properties, and evidence for two systemsJournal of Cellular Physiology, 1986
- Rate-limiting steps of 2-deoxyglucose uptake in rat adipocytesBiochimica et Biophysica Acta (BBA) - Biomembranes, 1980
- Phosphorylation but not transport of sugars is enhanced in virus-transformed mouse 3T3 cellsJournal of Cellular Physiology, 1975
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970