Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1991 Jan 1;273(Pt 1):49–56. doi: 10.1042/bj2730049

The alpha beta monomer of the insulin receptor has hormone-responsive tyrosine kinase activity.

E R Mortensen 1, J G Drachman 1, G Guidotti 1
PMCID: PMC1150211  PMID: 1998536

Abstract

Insulin receptors from turkey erythrocyte membranes exist as monomers and dimers when membranes are solubilized with detergent. We examined the ability of monomers and dimers to act as protein kinases to effect both autophosphorylation of the receptor and phosphorylation of an exogenous substrate. After separation by sucrose-density-gradient centrifugation, only receptor dimers show significant basal and insulin-stimulated kinase activity, whereas material at the position of receptor monomers is not active. Partial reduction of the membrane-bound receptors with dithiothreitol, however, produces a receptor monomer containing an alpha and a beta chain which has protein kinase activity similar to that of the original dimers. With rat adipocyte plasma membranes, which in the absence of reducing agents only contain receptor dimers, reduction with dithiothreitol also produces monomers with receptor kinase activity. Receptor monomer hormone-dependent kinase activity is insensitive to receptor concentration and shows stimulation after immobilization on an affinity support.

Full text

PDF
49

Images in this article

51Fig. 1.
on p.51
52Fig. 3.
on p.52
53Fig. 4.
on p.53
53Fig. 5.
on p.53
55Fig. 8.
on p.55
55Fig. 9.
on p.55

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aiyer R. A. Structural characterization of insulin receptors. I. Hydrodynamic properties of receptors from turkey erythrocytes. J Biol Chem. 1983 Dec 25;258(24):14992–14999. [PubMed] [Google Scholar]
  2. Aiyer R. A. Structural characterization of insulin receptors. II. Subunit composition of receptors from turkey erythrocytes. J Biol Chem. 1983 Dec 25;258(24):15000–15003. [PubMed] [Google Scholar]
  3. Böni-Schnetzler M., Kaligian A., DelVecchio R., Pilch P. F. Ligand-dependent intersubunit association within the insulin receptor complex activates its intrinsic kinase activity. J Biol Chem. 1988 May 15;263(14):6822–6828. [PubMed] [Google Scholar]
  4. Böni-Schnetzler M., Rubin J. B., Pilch P. F. Structural requirements for the transmembrane activation of the insulin receptor kinase. J Biol Chem. 1986 Nov 15;261(32):15281–15287. [PubMed] [Google Scholar]
  5. Cushman S. W., Wardzala L. J. Potential mechanism of insulin action on glucose transport in the isolated rat adipose cell. Apparent translocation of intracellular transport systems to the plasma membrane. J Biol Chem. 1980 May 25;255(10):4758–4762. [PubMed] [Google Scholar]
  6. Fujita-Yamaguchi Y., Choi S., Sakamoto Y., Itakura K. Purification of insulin receptor with full binding activity. J Biol Chem. 1983 Apr 25;258(8):5045–5049. [PubMed] [Google Scholar]
  7. Fujita-Yamaguchi Y., Kathuria S. The monomeric alpha beta form of the insulin receptor exhibits much higher insulin-dependent tyrosine-specific protein kinase activity than the intact alpha 2 beta 2 form of the receptor. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6095–6099. doi: 10.1073/pnas.82.18.6095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ginsberg B. H., Kahn C. R., Roth J. The insulin receptor of the turkey erythrocyte. Characterization of the membrane-bound receptor. Biochim Biophys Acta. 1976 Aug 16;443(2):227–242. doi: 10.1016/0005-2736(76)90506-x. [DOI] [PubMed] [Google Scholar]
  9. Hedo J. A., Kahn C. R., Hayashi M., Yamada K. M., Kasuga M. Biosynthesis and glycosylation of the insulin receptor. Evidence for a single polypeptide precursor of the two major subunits. J Biol Chem. 1983 Aug 25;258(16):10020–10026. [PubMed] [Google Scholar]
  10. Herrera R., Lebwohl D., Garcia de Herreros A., Kallen R. G., Rosen O. M. Synthesis, purification, and characterization of the cytoplasmic domain of the human insulin receptor using a baculovirus expression system. J Biol Chem. 1988 Apr 25;263(12):5560–5568. [PubMed] [Google Scholar]
  11. Jacobs S., Sahyoun N. E., Saltiel A. R., Cuatrecasas P. Phorbol esters stimulate the phosphorylation of receptors for insulin and somatomedin C. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6211–6213. doi: 10.1073/pnas.80.20.6211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kasuga M., Fujita-Yamaguchi Y., Blithe D. L., Kahn C. R. Tyrosine-specific protein kinase activity is associated with the purified insulin receptor. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2137–2141. doi: 10.1073/pnas.80.8.2137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kasuga M., Karlsson F. A., Kahn C. R. Insulin stimulates the phosphorylation of the 95,000-dalton subunit of its own receptor. Science. 1982 Jan 8;215(4529):185–187. doi: 10.1126/science.7031900. [DOI] [PubMed] [Google Scholar]
  14. Kasuga M., Zick Y., Blith D. L., Karlsson F. A., Häring H. U., Kahn C. R. Insulin stimulation of phosphorylation of the beta subunit of the insulin receptor. Formation of both phosphoserine and phosphotyrosine. J Biol Chem. 1982 Sep 10;257(17):9891–9894. [PubMed] [Google Scholar]
  15. Kohanski R. A., Frost S. C., Lane M. D. Insulin-dependent phosphorylation of the insulin receptor-protein kinase and activation of glucose transport in 3T3-L1 adipocytes. J Biol Chem. 1986 Sep 15;261(26):12272–12281. [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Massague J., Pilch P. F., Czech M. P. A unique proteolytic cleavage site on the beta subunit of the insulin receptor. J Biol Chem. 1981 Apr 10;256(7):3182–3190. [PubMed] [Google Scholar]
  18. Massagué J., Czech M. P. Role of disulfides in the subunit structure of the insulin receptor. Reduction of class I disulfides does not impair transmembrane signalling. J Biol Chem. 1982 Jun 25;257(12):6729–6738. [PubMed] [Google Scholar]
  19. McKeel D. W., Jarett L. Preparation and characterization of a plasma membrane fraction from isolated fat cells. J Cell Biol. 1970 Feb;44(2):417–432. doi: 10.1083/jcb.44.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Morrison B. D., Swanson M. L., Sweet L. J., Pessin J. E. Insulin-dependent covalent reassociation of isolated alpha beta heterodimeric insulin receptors into an alpha 2 beta 2 heterotetrameric disulfide-linked complex. J Biol Chem. 1988 Jun 5;263(16):7806–7813. [PubMed] [Google Scholar]
  21. Pike L. J., Eakes A. T., Krebs E. G. Characterization of affinity-purified insulin receptor/kinase. Effects of dithiothreitol on receptor/kinase function. J Biol Chem. 1986 Mar 15;261(8):3782–3789. [PubMed] [Google Scholar]
  22. Pilch P. F., Czech M. P. Interaction of cross-linking agents with the insulin effector system of isolated fat cells. Covalent linkage of 125I-insulin to a plasma membrane receptor protein of 140,000 daltons. J Biol Chem. 1979 May 10;254(9):3375–3381. [PubMed] [Google Scholar]
  23. Rosen O. M., Herrera R., Olowe Y., Petruzzelli L. M., Cobb M. H. Phosphorylation activates the insulin receptor tyrosine protein kinase. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3237–3240. doi: 10.1073/pnas.80.11.3237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Roth R. A., Cassell D. J. Insulin receptor: evidence that it is a protein kinase. Science. 1983 Jan 21;219(4582):299–301. doi: 10.1126/science.6849137. [DOI] [PubMed] [Google Scholar]
  25. Roth R. A., Mesirow M. L., Cassell D. J. Preferential degradation of the beta subunit of purified insulin receptor. Effect on insulin binding and protein kinase activities of the receptor. J Biol Chem. 1983 Dec 10;258(23):14456–14460. [PubMed] [Google Scholar]
  26. Schlessinger J. Signal transduction by allosteric receptor oligomerization. Trends Biochem Sci. 1988 Nov;13(11):443–447. doi: 10.1016/0968-0004(88)90219-8. [DOI] [PubMed] [Google Scholar]
  27. Shia M. A., Pilch P. F. The beta subunit of the insulin receptor is an insulin-activated protein kinase. Biochemistry. 1983 Feb 15;22(4):717–721. doi: 10.1021/bi00273a001. [DOI] [PubMed] [Google Scholar]
  28. Shia M. A., Rubin J. B., Pilch P. F. The insulin receptor protein kinase. Physicochemical requirements for activity. J Biol Chem. 1983 Dec 10;258(23):14450–14455. [PubMed] [Google Scholar]
  29. Sweet L. J., Morrison B. D., Wilden P. A., Pessin J. E. Insulin-dependent intermolecular subunit communication between isolated alpha beta heterodimeric insulin receptor complexes. J Biol Chem. 1987 Dec 5;262(34):16730–16738. [PubMed] [Google Scholar]
  30. Sweet L. J., Wilden P. A., Pessin J. E. Dithiothreitol activation of the insulin receptor/kinase does not involve subunit dissociation of the native alpha 2 beta 2 insulin receptor subunit complex. Biochemistry. 1986 Nov 4;25(22):7068–7074. doi: 10.1021/bi00370a047. [DOI] [PubMed] [Google Scholar]
  31. Ullrich A., Bell J. R., Chen E. Y., Herrera R., Petruzzelli L. M., Dull T. J., Gray A., Coussens L., Liao Y. C., Tsubokawa M. Human insulin receptor and its relationship to the tyrosine kinase family of oncogenes. 1985 Feb 28-Mar 6Nature. 313(6005):756–761. doi: 10.1038/313756a0. [DOI] [PubMed] [Google Scholar]
  32. Ullrich A., Schlessinger J. Signal transduction by receptors with tyrosine kinase activity. Cell. 1990 Apr 20;61(2):203–212. doi: 10.1016/0092-8674(90)90801-k. [DOI] [PubMed] [Google Scholar]
  33. Villalba M., Wente S. R., Russell D. S., Ahn J. C., Reichelderfer C. F., Rosen O. M. Another version of the human insulin receptor kinase domain: expression, purification, and characterization. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7848–7852. doi: 10.1073/pnas.86.20.7848. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. White M. F., Haring H. U., Kasuga M., Kahn C. R. Kinetic properties and sites of autophosphorylation of the partially purified insulin receptor from hepatoma cells. J Biol Chem. 1984 Jan 10;259(1):255–264. [PubMed] [Google Scholar]
  35. Yu K. T., Czech M. P. Tyrosine phosphorylation of the insulin receptor beta subunit activates the receptor-associated tyrosine kinase activity. J Biol Chem. 1984 Apr 25;259(8):5277–5286. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES