Characterization of GLUT3 protein expressed in Chinese hamster ovary cells

T. Asano, H. Katagiri, K. Takata, K. Tsukuda, J. L. Lin, H. Ishihara, K. Inukai, H. Hirano, Y. Yazaki, Y. Oka

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34 Citations (Scopus)

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 K(m) value of GLUT3 protein for 3-O-methylglucose uptake was approx 35% of that of GLUT1 protein, whereas the K(m) value of GLUT3 protein for 2-deoxy-D-glucose uptake was very similar to that of GLUT1 protein. The V(max) 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 K(m) and V(max) values for 3-O-methylglucose uptake.

Original languageEnglish
Pages (from-to)189-193
Number of pages5
JournalBiochemical Journal
Volume288
Issue number1
DOIs
Publication statusPublished - 1992

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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