title: Adipocyte insulin-binding species: the 40 A Stoke's radius protein.
abstract: Several laboratories have demonstrated the presence of large (70 A) (1 A = 0.1 nm) and small (40 A) insulin receptors. This report provides evidence that the 40 A insulin receptor migrates on dodecyl sulfate - acrylamide gel electrophoresis as a 90 000 dalton protein and that this protein is a single polypeptide chain. 125I-labeled insulin was bound to plasma membranes from isolated rat adipocytes. Following removal of unbound 125I-labeled insulin, the mixture was exposed to disuccinimidyl suberate. Proteins tagged with 125I-labeled insulin were separated by dodecyl sulfate gel electrophoresis or Sepharose 6B chromatography. Autoradiography of the gels demonstrated several large (relative mass (Mr) greater than 300 000) and one small (Mr approximately 90 000) labeled protein in nonreduced membrane proteins. Dithiothreitol reduction decreased the large insulin-binding species to its known subunits, but the 90 000 dalton protein did not decrease in size. Triton X-100 solubilized plasma membranes were separated by Sepharose 6B chromatography. One labeled protein, with Kav = 0.57 elution position, on dodecyl sulfate gel electrophoresis migrated as a 90 000 dalton protein. Thus, rat adipocyte plasma membranes contain both an oligomeric insulin-binding species and a monomeric insulin-binding species. The relationship of the monomeric to the oligomeric insulin receptor is discussed.
mesh_terms: D000273:Adipose Tissue / Q000032:analysis*; D000818:Animals; D002462:Cell Membrane / Q000032:analysis; D002850:Chromatography, Gel; D004229:Dithiothreitol / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D007328:Insulin / Q000378:metabolism; D046911:Macromolecular Substances; D008297:Male; D008433:Mathematics; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains; D011972:Receptor, Insulin / Q000032:analysis*
chemical_list: D007328:Insulin; D046911:Macromolecular Substances; D011972:Receptor, Insulin; D004229:Dithiothreitol