Evidence that functional erythrocyte-type glucose transporters are oligomers
Hebert, Daniel N
Harrison, Scott A.
Clancy, Brian M.
Buxton, Joanne M.
Czech, Michael P.
UMass Chan AffiliationsDepartment of Biochemistry and Molecular Pharmacology
Graduate School of Biomedical Sciences
Program in Molecular Medicine
Document TypeJournal Article
Keywords3T3 Cells; Animals; Blotting, Western; Cell Membrane; Cells, Cultured; Chimera; Cricetinae; Cricetulus; Erythrocytes; Humans; Mice; Monosaccharide Transport Proteins; Rats; Restriction Mapping; Transfection
Medicine and Health Sciences
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AbstractIn this study we tested the hypothesis that functional erythrocyte-type glucose transporters (GLUT1) exist as oligomeric complexes by expressing chimeric transporter proteins in Chinese hamster ovary cells harboring endogenous GLUT1 transporters. The chimeric transporters were GLUT1-4c, in which the 29 C-terminal residues of human GLUT1 were replaced by the 30 C-terminal residues of rat skeletal muscle glucose transporter (GLUT4), and GLUT1n-4, containing the N-terminal 199 residues of GLUT1 and the 294 C-terminal residues of GLUT4. Endogenous GLUT1 was quantitatively co-immunoprecipitated by using an anti-GLUT4 C-terminal peptide antibody from detergent extracts of Chinese hamster ovary cells expressing either of the chimeric proteins, as detected by immunoblotting the precipitates with an anti-GLUT1 C-terminal peptide antiserum. No co-immunoprecipitation of native GLUT1 with native GLUT4 from extracts of 3T3-L1 adipocytes, which contain both these transporters, was observed with the same antibody. These data are consistent with the hypothesis that GLUT1 transporters exist as homodimers or higher order oligomers and that a major determinant of oligomerization is located within the first 199 residues of GLUT1.
J Biol Chem. 1991 Oct 25;266(30):20213-7.