Identification and partial purification of the insulin-responsive glucose transporter from 3T3-L1 adipocytes.
The glucose transporter in 3T3-L1 adipocytes has been identified as a polypeptide of average Mr 51000 by means of its reaction with antibodies raised against the purified human erythrocyte glucose transporter and by photolabeling with [3H]cytochalasin B. The finding that the antibodies immunoprecipitated the photolabeled polypeptide demonstrated that both methods detected the same polypeptide. The 3T3-L1 adipocyte glucose transporter has been partially purified. The main steps in the purification procedure were the preparation of salt-washed cellular membranes, Triton X-100 solubilization, and immunoaffinity chromatography on affinity-purified antibodies against the human erythrocyte transporter. A simple method of affinity purification of these antibodies, which consists of adsorption from serum onto protein-depleted erythrocyte membranes and release with acid, and an assay for the 3T3-L1 adipocyte transporter polypeptide, which employs immunoblotting, have been developed.
D000273:Adipose Tissue / Q000032:analysis* / Q000166:cytology; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000032:analysis; D002846:Chromatography, Affinity; D007328:Insulin / Q000494:pharmacology; D051379:Mice; D009004:Monosaccharide Transport Proteins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*
D007328:Insulin; D009004:Monosaccharide Transport Proteins