Purification and properties of the human placental insulin receptor

J Biol Chem. 1981 Sep 10;256(17):9266-73.

Abstract

Human placenta insulin receptor was purified 11,000-fold to near homogeneity using DEAE-cellulose chromatography and affinity chromatography on insulin-Sepharose. Approximately 200 to 300 micrograms of purified receptor were obtained from a single placenta. In solution, the native receptor is a complex (Mr = 440,000) of an acidic, multi-subunit protein with a Mr of 350,000 and bound detergent accounting for the remainder of the mass. The receptor protein is asymmetric (f/f0 = 1.4) and consists of a single Coomassie blue staining polypeptide of Mr = 135,000. In addition to the 135,000-dalton polypeptide, two smaller polypeptides of Mr = 45,000 and 90,000 were observed upon autoradiography of 125I-labeled receptor subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These two smaller polypeptides did not stain with Coomassie blue but migrated with native receptor activity on isoelectric focusing gels and were coimmunoprecipitated with the 135,000-dalton polypeptide by anti-insulin receptor antibody. The 135,000-dalton subunit was specifically labeled by 125I-insulin using the bifunctional cross-linking reagent disuccinimidyl suberate (P. F. Pilch, and M. P. Czech, (1979) J. Biol. Chem. 254, 3375-3381), suggesting that this component contains the insulin binding domain.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Binding, Competitive
  • Cell Membrane / metabolism
  • Female
  • Humans
  • Insulin / analogs & derivatives
  • Insulin / metabolism
  • Kinetics
  • Macromolecular Substances
  • Molecular Weight
  • Placenta / metabolism*
  • Pregnancy
  • Protein Conformation
  • Receptor, Insulin / isolation & purification*
  • Receptor, Insulin / metabolism

Substances

  • Insulin
  • Macromolecular Substances
  • insulin, iodo-
  • Receptor, Insulin