Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1988 Feb 15;250(1):95–101. doi: 10.1042/bj2500095

Hydrogen peroxide stimulates tyrosine phosphorylation of the insulin receptor and its tyrosine kinase activity in intact cells.

O Koshio 1, Y Akanuma 1, M Kasuga 1
PMCID: PMC1148820  PMID: 2833239

Abstract

H-35 rat hepatoma cells were labelled with [32P]orthophosphate and their insulin receptors isolated on wheat germ agglutinin (WGA)-agarose and anti-(insulin receptor) serum. The incubation of these cells with 10 mM-H2O2 for 10 min increased the phosphorylation of both the serine and tyrosine residues of the beta subunit of the insulin receptor. Next, insulin receptors were purified on WGA-agarose from control and H2O2-treated H-35 cells and the purified fractions incubated with [gamma-32P]ATP and Mn2+. Phosphorylation of the beta subunit of insulin receptors obtained from H2O2-treated cells was 150% of that of control cells. The kinase activity of the WGA-purified receptor preparation obtained from H2O2-treated cells, as measured by phosphorylation of src-related synthetic peptide, was increased about 4-fold over control cells. These data suggest that in intact cell systems, H2O2 may increase the insulin receptor kinase activity by inducing phosphorylation of the beta subunit of insulin receptor.

Full text

PDF
95

Images in this article

Selected References

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

  1. Cuatrecasas P., Tell G. P. Insulin-like activity of concanavalin A and wheat germ agglutinin--direct interactions with insulin receptors. Proc Natl Acad Sci U S A. 1973 Feb;70(2):485–489. doi: 10.1073/pnas.70.2.485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Czech M. P., Lawrence J. C., Jr, Lynn W. S. Evidence for the involvement of sulfhydryl oxidation in the regulation of fat cell hexose transport by insulin. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4173–4177. doi: 10.1073/pnas.71.10.4173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dubyak G. R., Kleinzeller A. The insulin-mimetic effects of vanadate in isolated rat adipocytes. Dissociation from effects of vanadate as a (Na+-K+)ATPase inhibitor. J Biol Chem. 1980 Jun 10;255(11):5306–5312. [PubMed] [Google Scholar]
  4. Ebina Y., Ellis L., Jarnagin K., Edery M., Graf L., Clauser E., Ou J. H., Masiarz F., Kan Y. W., Goldfine I. D. The human insulin receptor cDNA: the structural basis for hormone-activated transmembrane signalling. Cell. 1985 Apr;40(4):747–758. doi: 10.1016/0092-8674(85)90334-4. [DOI] [PubMed] [Google Scholar]
  5. Ellis L., Clauser E., Morgan D. O., Edery M., Roth R. A., Rutter W. J. Replacement of insulin receptor tyrosine residues 1162 and 1163 compromises insulin-stimulated kinase activity and uptake of 2-deoxyglucose. Cell. 1986 Jun 6;45(5):721–732. doi: 10.1016/0092-8674(86)90786-5. [DOI] [PubMed] [Google Scholar]
  6. Kadowaki T., Kasuga M., Akanuma Y., Ezaki O., Takaku F. Decreased autophosphorylation of the insulin receptor-kinase in streptozotocin-diabetic rats. J Biol Chem. 1984 Nov 25;259(22):14208–14216. [PubMed] [Google Scholar]
  7. Kahn C. R., Baird K., Filier J. S., Jarrett D. B. Effects of autoantibodies to the insulin receptor on isolated adipocytes. Studies of insulin binding and insulin action. J Clin Invest. 1977 Nov;60(5):1094–1106. doi: 10.1172/JCI108861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kasuga M., Akanuma Y., Tsushima T., Suzuki K., Kosaka K., Kibata M. Effects of antiinsulin receptor autoantibody on the metabolism of rat adipocytes. J Clin Endocrinol Metab. 1978 Jul;47(1):66–77. doi: 10.1210/jcem-47-1-66. [DOI] [PubMed] [Google Scholar]
  9. Kasuga M., Fujita-Yamaguchi Y., Blithe D. L., White M. F., Kahn C. R. Characterization of the insulin receptor kinase purified from human placental membranes. J Biol Chem. 1983 Sep 25;258(18):10973–10980. [PubMed] [Google Scholar]
  10. 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]
  11. Kasuga M., Zick Y., Blithe D. L., Crettaz M., Kahn C. R. Insulin stimulates tyrosine phosphorylation of the insulin receptor in a cell-free system. Nature. 1982 Aug 12;298(5875):667–669. doi: 10.1038/298667a0. [DOI] [PubMed] [Google Scholar]
  12. Klein H. H., Freidenberg G. R., Kladde M., Olefsky J. M. Insulin activation of insulin receptor tyrosine kinase in intact rat adipocytes. An in vitro system to measure histone kinase activity of insulin receptors activated in vivo. J Biol Chem. 1986 Apr 5;261(10):4691–4697. [PubMed] [Google Scholar]
  13. 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]
  14. Le Marchand-Brustel Y., Grémeaux T., Ballotti R., Van Obberghen E. Insulin receptor tyrosine kinase is defective in skeletal muscle of insulin-resistant obese mice. Nature. 1985 Jun 20;315(6021):676–679. doi: 10.1038/315676a0. [DOI] [PubMed] [Google Scholar]
  15. Livingston J. N., Gurny P. A., Lockwood D. H. Insulin-like effects of polyamines in fat cells. Mediation by H2O2 formation. J Biol Chem. 1977 Jan 25;252(2):560–562. [PubMed] [Google Scholar]
  16. Lockwood D. H., East L. E. Studies of the insulin-like actions of polyamines on lipid and glucose metabolism in adipose tissue cells. J Biol Chem. 1974 Dec 25;249(24):7717–7722. [PubMed] [Google Scholar]
  17. Massague J., Pilch P. F., Czech M. P. Electrophoretic resolution of three major insulin receptor structures with unique subunit stoichiometries. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7137–7141. doi: 10.1073/pnas.77.12.7137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. May J. M., de Haën C. The insulin-like effect of hydrogen peroxide on pathways of lipid synthesis in rat adipocytes. J Biol Chem. 1979 Sep 25;254(18):9017–9021. [PubMed] [Google Scholar]
  19. 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]
  20. Tamura S., Brown T. A., Whipple J. H., Fujita-Yamaguchi Y., Dubler R. E., Cheng K., Larner J. A novel mechanism for the insulin-like effect of vanadate on glycogen synthase in rat adipocytes. J Biol Chem. 1984 May 25;259(10):6650–6658. [PubMed] [Google Scholar]
  21. 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]
  22. Van Obberghen E., Kowalski A. Phosphorylation of the hepatic insulin receptor: stimulating effect of insulin on intact cells and in a cell-free system. FEBS Lett. 1982 Jul 5;143(2):179–182. doi: 10.1016/0014-5793(82)80094-x. [DOI] [PubMed] [Google Scholar]
  23. Van Obberghen E., Ksauga M., Le Cam A., Hedo J. A., Itin A., Harrison L. C. Biosynthetic labeling of insulin receptor: studies of subunits in cultured human IM-9 lymphocytes. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1052–1056. doi: 10.1073/pnas.78.2.1052. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Yu K. T., Czech M. P. Tyrosine phosphorylation of insulin receptor beta subunit activates the receptor tyrosine kinase in intact H-35 hepatoma cells. J Biol Chem. 1986 Apr 5;261(10):4715–4722. [PubMed] [Google Scholar]

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

RESOURCES