Nutrient control of GLUT1 processing and turnover in 3T3-L1 adipocytes.
Metabolic labeling and immunoprecipitation were used to analyze the glucose-dependent regulation of GLUT1 synthesis, processing, and turnover in a murine adipocyte cell line. Metabolically labeled GLUT1 from control cells migrated as a 46-kDa protein, while GLUT1 from cells deprived of glucose for more than 12 h migrated as a 37-kDa protein. On the basis of tunicamycin sensitivity, both GLUT1 species arose from a common protein migrating at 36 kDa. In addition, the rate of synthesis of GLUT1 in control and glucose-deprived cells was similar. In short pulse-chase experiments, we distinguished two species arising from the core GLUT1 protein in control cells; an intermediate and the mature 46-kDa species. In contrast, only one glycoform, the 37-kDa species, arose from the core protein in glucose-deprived cells, which was not further processed in either the presence or absence of glucose. Although 12-18 h of glucose deprivation were required to affect GLUT1 glycosylation, glucose-deprived cells quickly recovered the ability to correctly glycosylate GLUT1 upon the readdition of glucose (t1/2 < 1 h). GLUT1 in control adipocytes exhibited a half-life of approximately 14 h, while that in glucose-deprived adipocytes was greater than 50 h. This effect was readily reversed upon the readdition of glucose. In total, these data show that glucose deprivation alters both the processing (glycosylation) and turnover (degradation) of GLUT1. These results are discussed in light of transport function.
McMahon
R J
RJ
Department of Biochemistry and Molecular Biology, University of Florida, Gainesville 32610, USA.
Frost
S C
SC
D016475:3T3 Cells / Q000187:drug effects / Q000378:metabolism*; D000273:Adipose Tissue / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D004734:Energy Metabolism; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D051272:Glucose Transporter Type 1; D006031:Glycosylation; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009004:Monosaccharide Transport Proteins / Q000276:immunology / Q000378:metabolism*; D010446:Peptide Fragments / Q000276:immunology; D011233:Precipitin Tests; D011499:Protein Processing, Post-Translational
D051272:Glucose Transporter Type 1; D009004:Monosaccharide Transport Proteins; D010446:Peptide Fragments; C495678:Slc2a1 protein, mouse; D005947:Glucose