title: C-terminal mutations that alter the turnover number for 3-O-methylglucose transport by GLUT1 and GLUT4.
abstract: Turnover numbers for 3-O-methylglucose transport by the homologous glucose transporters GLUT1 and GLUT4 were compared to those for truncated and chimeric transporters expressed in Xenopus oocytes to assess potential regulatory properties of the C-terminal domain. The ability of high intracellular sugar concentrations to increase the turnover number for sugar entry ("accelerated exchange") by GLUT1 and not by GLUT4 was maintained in oocytes. Replacing the GLUT1 C terminus with that of GLUT4 stimulated turnover 1.6-fold, but abolished accelerated exchange. Thus, the GLUT1 C terminus permits accelerated exchange by GLUT1, but in doing so must interact with other GLUT1 specific sequences since the GLUT4ctrm1 chimera did not exhibit this kinetic property. Removal of 38 C-terminal amino acids from GLUT4 reduced its turnover number by 40%, whereas removing only 20 residues or replacing its C terminus with that of GLUT1 increased its turnover number 3.5-3.9 fold. Therefore, using mechanisms independent of those which alter transporter targeting to the plasma membrane, C-terminal mutations in either GLUT1 or GLUT4 can activate transport normally restricted by the native C-terminal domain. These results implicate the C termini as targets of physiological factors, which through covalent modification or direct binding might alter C-terminal interactions to regulate intrinsic GLUT1 and GLUT4 transporter activity.
authors.affiliation: Department of Biochemistry and Molecular Biology, Louisiana State University School of Medicine, Shreveport 71130, USA.
authors.forename: R
authors.initials: R
authors.lastname: Dauterive
authors.forename: S
authors.initials: S
authors.lastname: Laroux
authors.forename: R C
authors.initials: RC
authors.lastname: Bunn
authors.forename: A
authors.initials: A
authors.lastname: Chaisson
authors.forename: T
authors.initials: T
authors.lastname: Sanson
authors.forename: B C
authors.initials: BC
authors.lastname: Reed
mesh_terms: D019325:3-O-Methylglucose; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D017124:Conserved Sequence; D005260:Female; D051272:Glucose Transporter Type 1; D051275:Glucose Transporter Type 4; D007700:Kinetics; D008757:Methylglucosides / Q000378:metabolism*; D051379:Mice; D008961:Models, Structural; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D009124:Muscle Proteins; D016296:Mutagenesis; D009865:Oocytes / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology; D014176:Protein Biosynthesis; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic; D014982:Xenopus laevis
chemical_list: D000906:Antibodies; D051272:Glucose Transporter Type 1; D051275:Glucose Transporter Type 4; D008757:Methylglucosides; D009004:Monosaccharide Transport Proteins; D009124:Muscle Proteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C495678:Slc2a1 protein, mouse; C495693:Slc2a4 protein, mouse; D019325:3-O-Methylglucose