Internalization of Insulin: Structures Involved and Significance

  • Barry I. Posner
  • Masood N. Kahn
  • John J. M. Bergeron
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 189)


The binding of insulin to its receptor is followed by aggregation of hormone-receptor complexes and their internalization into the cell. Internalized hormone is concentrated in Golgi-enriched not lysosomal endocytotic structures which, in rat liver, contain lipoprotein particles and can be resolved by centrifugation techniques into three different entities. Recent work has shown that the bulk of endocytotic structures can be resolved from biochemically defined (i.e., galactosyltransferase-containing) Golgi elements. The endosomal apparatus or endosomes appear to function as a sorting center wherein internalized hormone-receptor complexes are concentrated and dissociated prior to directing hormone to lysosomes and receptor back to the cell surface for reutilization. Endosomes are heterogeneous and different functions might be subserved by different endosomal structures. Since an insulin stimulable receptor kinase activity can be identified in endosomes certain aspects of insulin action might be initiated herein.


Insulin Receptor Peptide Hormone Percoll Gradient Centrifugation Golgi Element Cell BioI 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    A.D. Attie, R.C. Pittman, and D. Steinberg, Hepatic catabolism of low density lipoprotein: mechanisms and metabolic consequences, Hepatology, 2: 269–281 (1982).PubMedCrossRefGoogle Scholar
  2. 2.
    S.K. Basu, J.K. Goldstein, R.G.W. Anderson, and M.S. Brown, Monensin interrupts the recycling of low density lipoprotein receptors in human fibroblasts, Cell, 24: 493–502 (1981).PubMedCrossRefGoogle Scholar
  3. 3.
    J.J.M. Bergeron, J.H. Ehrenreich, P. Siekevitz, and G.E. Palade, Golgi fractions prepared from rat liver homogenates: isolation procedure and morphological characterization, J. Cell Biol, 59: 73–88 (1973).PubMedCrossRefGoogle Scholar
  4. 4.
    S.J.M. Bergeron, L. Resch, R. Rachubinski, B. Patel, and B.I. Posner, Effect of colchicine on internalization of prolactin in female rat liver: an in vivo radioautographic study, J. Cell Biol, 96: 875–886 (1983).PubMedCrossRefGoogle Scholar
  5. 5.
    P. Bernahu, J.H. Olefsky, P. Tsai, P. Thamm, D. Saunders, and D. Brandenburg, Internalization and molecular processing of insulin receptors in isolated rat adipocytes, Proc Natl. Acad. Sci. U.S.A. 79: 4069–4073 (1982).CrossRefGoogle Scholar
  6. 6.
    M.S. Brown, and J.L. Goldstein, Receptor-mediated endocytosis: insights from the lipoprotein receptor system, Proc. Natl. Acad. Sci. U.S.A. 76: 3330–3337 (1979).PubMedCrossRefGoogle Scholar
  7. 7.
    Y. S. Chao, A.L. Jones, G.T. Hradek, E.E.T. Windier, and R.J. Havel, Autoradiographic localization of the sites of uptake, cellular transport, and catobolism of low density lipoproteins in the liver of normal and estrogen-treated rats, Proc. Natl. Acad. Sci. U.S.A. 78: 597–601 (1981).PubMedCrossRefGoogle Scholar
  8. 8.
    P.J. Courtoy, J. Quintart, and P. Baudhuin, Shift of equilibrium density induced by 3, 3-diaminobenzidine cytochemistry: a new procedure for the analysis and purification of peroxidase-containing organelles, J. Cell Biol, 98: 870–876 (1984).PubMedCrossRefGoogle Scholar
  9. 9.
    J. Cruz, B.I. Posner, and J.J.H. Bergeron, Receptor-mediated endocytosis of 125I-Insulin into pancreatic acinar cells in vivo, Endocrinology 115: 1996–2008 (1984).PubMedCrossRefGoogle Scholar
  10. 10.
    R.A. Defronzo, and E. Ferrannini, The pathogenesis of noninsulin-dependent diabetes: an update, Medicine, 61: 125–140 (1982).PubMedCrossRefGoogle Scholar
  11. 11.
    C. de Haen, The non-stoichiometric floating receptor model for hormone-sensitive adenylyl cyclase, J. Theor. Biol. 58: 383–400 (1976).PubMedCrossRefGoogle Scholar
  12. 12.
    B. Desbuquois, S. Lopez, and H. Burlet, Ligand-induced translocation of insulin receptors in intact rat liver, J. Biol. Chem. 257: 10852–10860 (1982).PubMedGoogle Scholar
  13. 13.
    R.B. Dickson, M.C. Willingham, and I. Pastan, a2-Macroglobulin adsorbed to colloidal gold: a new probe in the study of receptor-mediated endocytosis, J. Cell Biol, 89: 29–34 (1981).PubMedCrossRefGoogle Scholar
  14. 14.
    J.H. Ehrenreich, J.J.M. Bergeron, P. Siekevitz, and G.E. Palade, Golgi fractions prepared from rat liver homogenates: isolation procedure and morphological characterization, J. Cell Biol. 59: 45–72 (1973).PubMedCrossRefGoogle Scholar
  15. 15.
    C.A. Enns, J.W. Larrick, H. Suomalainen, J. Schroder, and H.H. Sussman, Co-migration and internalization of transferrin and its receptor on K562 cells, J. Cell Biol. 97: 579–585 (1983).PubMedCrossRefGoogle Scholar
  16. 16.
    M. Fehlmann, J.-L. Carpentier, A. Le Cam, P. Thamm, D. Saunders, D. Brandenburg, L. Orci, and P. Freychet, Biochemical and morphological evidence that the insulin receptor is internalized with insulin in hepatocytes, J. Cell Biol. 93: 82–87 (1982).PubMedCrossRefGoogle Scholar
  17. 17.
    M. Fehlmann, J.L. Carpentier, E. Van Obberghen, P. Freychet, P. Thamm, D. Saunders, D. Brandenburg, and L. Orci, Internalized insulin receptors are recycled to the cell surface in rat hepatocytes, Proc. Natl. Acad. Sci. USA, 79: 5921–5925 (1982).PubMedCrossRefGoogle Scholar
  18. 18.
    H.J. Geuze, J.W. Slot, G.J.A.M. Strous, H.F. Lodish, and A.L. Schwartz, Intracellular site of asialoglycoprotein receptor-ligand uncoupling: double-label immunoelectron microscopy during receptor-mediated endocytosis, Cell 32: 277–287 (1983).PubMedCrossRefGoogle Scholar
  19. 19.
    J.J. Geuze, J.W. Slot, G.J.A.M. Strous, J. Peppard, K. von Figura, A. Hasilik, and A.L. Schwartz, Intracellular receptor sorting during endocytosis: comparative immunoelectron microscopy of multiple receptors in rat liver, Cell, 37: 195–204 (1984).PubMedCrossRefGoogle Scholar
  20. 20.
    P. Gorden, J.-L. Carpentier, P. Freychet, and L. Orci, Internalization of polypeptide hormones: mechanism, intracellular localization and significance, Diabetologia, 18: 263–274 (1980).PubMedCrossRefGoogle Scholar
  21. 21.
    A. Green, and J.K. Olefsky, Evidence for insulin-induced internalization and degradation of insulin receptors in rat adipocytes, Proc. Natl. Acad. Sci. USA, 79: 427–431 (1982).PubMedCrossRefGoogle Scholar
  22. 22.
    A. Helenius, J. Hellman, D. Wall, and A. Hubbard, Endosomes, Trends Biochem. Sci. 7: 245–250 (1983).CrossRefGoogle Scholar
  23. 23.
    C.R. Hopkins, Intracellular routing of transferrin and transferrin receptors in epidermoid carcinoma A431 cells, Cell, 32: 321–330 (1983).CrossRefGoogle Scholar
  24. 24.
    S. Jacobs, and P. Cuatrecasas, The mobile receptor hypothesis and cooperativity of hormone binding: application to insulin, Biochim. Biophys. Acta, 433: 482–495 (1976).PubMedCrossRefGoogle Scholar
  25. 25.
    M. Kasuga, Y. Fujita-Yamaguchi, D.L. Blithe, H.F. White, and C.R. Kahn, Characterization of the insulin receptor kinase purified from human placental membranes, J. Biol. Chem. 258: 10973–10980 (1983).PubMedGoogle Scholar
  26. 26.
    D.G. Kay, H.N. Khan, B.I. Posner, and J.J.K. Bergeron, 125I-insulin in hepatic Golgi fractions: application of the diaminobenzidine (DAB)-shift protocol, Biochem. Biophys. Res. Commun. 123: 1144–1148 (1984).PubMedCrossRefGoogle Scholar
  27. 27.
    H.N. Khan, B.I. Posner, A.K. Verma, R.J. Khan, and J.J.H. Bergeron, Intracellular hormone receptors: evidence for insulin and lactogen receptors in a unique vesicle sedimenting in lysosome fractions of rat liver, Proc. Natl. Acad. Sci. USA, 78: 4980–4984 (1981).PubMedCrossRefGoogle Scholar
  28. 28.
    H.N. Khan, B.I. Posner, R.J. Khan, and J.J.H. Bergeron, Internalization of insulin into rat liver Golgi elements: evidence for vesicle heterogeneity and the path of intracellular processing, J. Biol. Chem. 257: 5969–5976 (1982).PubMedGoogle Scholar
  29. 29.
    H.N. Khan, S. Savoie, J.J.H. Bergeron, and B.I. Posner, Endosomes of rat liver: preparation and characterization, Abstract, Amer. Soc. Cell Biol. Meeting (1984).Google Scholar
  30. 30.
    R.J. Khan, M.N. Khan, J.J.M. Bergeron, and B.I. Posner, Prolactin internalization into rat differential effects of chloroquine. (in press).Google Scholar
  31. 31.
    K. Kikuchi, J. Lerner, R. J. insulin fragments on biological activity of insulin and desoctapeptide insulin, I. Potentiation of biological activities, J. Biol. Chem. 256: 9441–9444 (1981).Google Scholar
  32. 32.
    M. Krupp, and M.D. Lane, On the mechanism of ligand-induced down-regulation of insulin receptor level in the liver cell, J. Biol. Chem. 256: 1689–1694 (1981).PubMedGoogle Scholar
  33. 33.
    L.C. Kuhn and J.P. Kraehenbuhl, The sacrificial receptor-translocation of polymeric IgA across epithelia, Trends Biochem Sci. 7: 299–302 (1982).CrossRefGoogle Scholar
  34. 34.
    J. Larner, G. Galasko, K. Cheng, A.A. Depaoli-Roach, L. Huang, P. Daggy and J. Kellogg, Generation by insulin of a chemical mediator that controls protein phosphorylation and dephosphorylation, Science 206: 1408–1410 (1979).PubMedCrossRefGoogle Scholar
  35. 35.
    J.A. McKanna, H.T. Haigler, and S. Cohen, Hormone receptor topology and dynamics: morphological analysis using ferritinlabeled epidermal growth factor, Proc. Natl. Acad. Sci. USA 76: 5689–5693 (1979).PubMedCrossRefGoogle Scholar
  36. 36.
    I.S. Hellman, H. Plutner, R.H. Steinman, J.C. Unkeless and Z.A. Cohn, J. Cell Biol. 96: 887–895 (1983).CrossRefGoogle Scholar
  37. 37.
    V. Pezzino, R. Vigneri, N.B. Pliam, and I.D. Goldfine, Rapid regulation of plasma membrane insulin receptors, Diabetologia 19: 211–215 (1980).PubMedCrossRefGoogle Scholar
  38. 38.
    B.I. Posner, J.J.M. Bergeron, Z. Josefsberg, M.N. Khan, R.J. Khan, B.A. Patel, R.A. Sikstrom, A.K. Verma, Polypeptide hormones: intracellular receptors and internalization, Recent Prog. Norm. Res. 37: 539–582 (1981).Google Scholar
  39. 39.
    B.I. Posner, M.N. Khan and J.J.M. Bergeron, Endocytosis of peptide hormones and other ligands, Endocr. Revs. 3: 280–298 (1982).CrossRefGoogle Scholar
  40. 40.
    B.I. Posner, B. Patel, M.N. Khan, J.J.H. Bergeron, chloroquine on the internalization of 1251_ insulin cellular fractions of rat liver: evidence for an chloroquine on Golgi elements, J. Biol. Chem. 257: 5789–5799 (1982).PubMedGoogle Scholar
  41. 41.
    G.H. Reaven, Insulin resistance in noninsulin-dependent diabetes mellitus. Does it exist and can it be measured? Amer J. Med 74 (No. 1A): 3–17 (1983)PubMedCrossRefGoogle Scholar
  42. 42.
    D.-J. Reijngoud, and J.H. Tager, Chloroquine accumulation isolated rat liver lysosomes, FEBS Lett 64: 231–235 (1976).PubMedCrossRefGoogle Scholar
  43. 43.
    Y. Schecter, J. Schlessinger, S. Jacobs, K-J Chang, and Cuatrecasas. Fluorescent labeling of hormone receptors in viable cells: preparation and properties of highly fluorescent derivatives of epidermal growth factor and insulin, Proc. Natl. Acad. Sci. USA 75: 2135–2139 (1978).CrossRefGoogle Scholar
  44. 44.
    S.J. Singer, and G.L. Nicolson, The fluid mosaic model of the structure of cell membranes, Science 175: 720–731 (1972).PubMedCrossRefGoogle Scholar
  45. 45.
    G.D. Smith and T.J. Peters, The localization in rat liver of alkaline phosphodiesterase to a discrete organelle implicated in ligand internalization, Biochim. Biophys. Acta 716: 24–30 (1982).PubMedCrossRefGoogle Scholar
  46. 46.
    S. Terris, and D.F. Steiner, Binding and degradation of 1251-insulin by rat hepatocytes, J. Biol. Chem. 250: 8389–8398 (1975).PubMedGoogle Scholar
  47. 47.
    B. Tycko and F.R. Maxfield, Rapid acidification of endocytic vesicles containing a2-macroglobulin, Cell 28: 643–651 (1982).PubMedCrossRefGoogle Scholar
  48. 48.
    D.A. Wall, G. Wilson, and A. Hubbard, The galactose specific recognition system of mammalian liver: the route of ligand internalization in rat hepatocytes, Cell, 21: 79–93 (1980).PubMedCrossRefGoogle Scholar
  49. 49.
    R.J. Walsh, B.I. Posner, and B. Patel, Binding and uptake of 1251-iodoprolactin by epithelial cells of rat choroids plexus: an in vivo autoradiographic analysis, Endocrinology, 114: 1496–1505 (1984).PubMedCrossRefGoogle Scholar
  50. 50.
    M.C. Willingham, and I. Pastan, The receptosome: an inter- mediate organelle of receptor-mediated endocytosis in cultured fibroblasts, Cell 21: 67–77 (1980).PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1985

Authors and Affiliations

  • Barry I. Posner
    • 1
  • Masood N. Kahn
    • 1
  • John J. M. Bergeron
    • 2
  1. 1.Department of MedicineThe Royal Victoria Hospital and McGill UniversityMontrealCanada
  2. 2.Department of AnatomyThe Royal Victoria Hospital and McGill UniversityMontrealCanada

Personalised recommendations