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. 1978 Oct;75(10):5025–5029. doi: 10.1073/pnas.75.10.5025

Epidermal growth factor: morphological demonstration of binding, internalization, and lysosomal association in human fibroblasts.

P Gorden, J L Carpentier, S Cohen, L Orci
PMCID: PMC336255  PMID: 311005

Abstract

Using a quantitative electron microscopic autoradiographic technique, we have localized the initial binding step of 125I-labeled epidermal growth factor (125I-EGF) to the plasma membrane of the human fibroblast. After initial binding, labeled EGF is internalized progressively by the cell in a time- and temperature-dependent fashion; when cell-associated radioactivity comes to steady state, approximately 1/3 of the autoradiographic grains are related to the plasma membrane and approximately 2/3 have been internalized. Under these conditions the internalized grains are almost exclusively related to lysosomal structures. When 125I-EGF associates with the cells for 2 hr at 4 degrees or 2 min at 37 degrees, 34% of grains localize to coated regions of the membrane. These coated regions make up less than 2% of the membrane surface. These data directly confirm kinetic studies and suggest that saturable binding of EGF is followed by adsorptive pinocytosis and cellular degradation of the ligand and possibly its cell surface receptor.

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Selected References

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

  1. Anderson R. G., Brown M. S., Goldstein J. L. Role of the coated endocytic vesicle in the uptake of receptor-bound low density lipoprotein in human fibroblasts. Cell. 1977 Mar;10(3):351–364. doi: 10.1016/0092-8674(77)90022-8. [DOI] [PubMed] [Google Scholar]
  2. Anderson R. G., Goldstein J. L., Brown M. S. Localization of low density lipoprotein receptors on plasma membrane of normal human fibroblasts and their absence in cells from a familial hypercholesterolemia homozygote. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2434–2438. doi: 10.1073/pnas.73.7.2434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. COHEN S. Isolation of a mouse submaxillary gland protein accelerating incisor eruption and eyelid opening in the new-born animal. J Biol Chem. 1962 May;237:1555–1562. [PubMed] [Google Scholar]
  4. Carpenter G., Cohen S. 125I-labeled human epidermal growth factor. Binding, internalization, and degradation in human fibroblasts. J Cell Biol. 1976 Oct;71(1):159–171. doi: 10.1083/jcb.71.1.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carpenter G., Lembach K. J., Morrison M. M., Cohen S. Characterization of the binding of 125-I-labeled epidermal growth factor to human fibroblasts. J Biol Chem. 1975 Jun 10;250(11):4297–4304. [PubMed] [Google Scholar]
  6. Carpentier J. L., Gorden P., Amherdt M., Van Obberghen E., Kahn C. R., Orci L. 125I-insulin binding to cultured human lymphocytes. Initial localization and fate of hormone determined by quantitative electron microscopic autoradiography. J Clin Invest. 1978 Apr;61(4):1057–1070. doi: 10.1172/JCI109005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cohen S., Carpenter G. Human epidermal growth factor: isolation and chemical and biological properties. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1317–1321. doi: 10.1073/pnas.72.4.1317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gavin J. R., 3rd, Roth J., Neville D. M., Jr, de Meyts P., Buell D. N. Insulin-dependent regulation of insulin receptor concentrations: a direct demonstration in cell culture. Proc Natl Acad Sci U S A. 1974 Jan;71(1):84–88. doi: 10.1073/pnas.71.1.84. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gorden P., Carpentier J. L., Cohen S., Orci L. Interaction du facteur de croissance épidermique avec des fibroblastes humains en culture : une étude autoradiographique à l'échelon ultrastructural. C R Acad Sci Hebd Seances Acad Sci D. 1978 May 22;286(20):1471–1474. [PubMed] [Google Scholar]
  10. Gorden P., Carpentier J. L., Freychet P., LeCam A., Orci L. Intracellular translocation of iodine-125-labeled insulin: direct demonstration in isolated hepatocytes. Science. 1978 May 19;200(4343):782–785. doi: 10.1126/science.644321. [DOI] [PubMed] [Google Scholar]
  11. Hollenberg M. D., Cuatrecasas P. Epidermal growth factor: receptors in human fibroblasts and modulation of action by cholera toxin. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2964–2968. doi: 10.1073/pnas.70.10.2964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Nadler N. J. The interpretation of grain counts in electron microscope radioautography. J Cell Biol. 1971 Jun;49(3):877–882. [PubMed] [Google Scholar]
  13. Orci L., Carpentier J. L., Perrelet A., Anderson R. G., Goldstein J. L., Brown M. S. Occurrence of low density lipoprotein receptors within large pits on the surface of human fibroblasts as demonstrated by freeze-etching. Exp Cell Res. 1978 Apr;113(1):1–13. doi: 10.1016/0014-4827(78)90081-2. [DOI] [PubMed] [Google Scholar]
  14. Pearse B. M. Clathrin: a unique protein associated with intracellular transfer of membrane by coated vesicles. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1255–1259. doi: 10.1073/pnas.73.4.1255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. ROTH T. F., PORTER K. R. YOLK PROTEIN UPTAKE IN THE OOCYTE OF THE MOSQUITO AEDES AEGYPTI. L. J Cell Biol. 1964 Feb;20:313–332. doi: 10.1083/jcb.20.2.313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Silverstein S. C., Steinman R. M., Cohn Z. A. Endocytosis. Annu Rev Biochem. 1977;46:669–722. doi: 10.1146/annurev.bi.46.070177.003321. [DOI] [PubMed] [Google Scholar]
  17. Terris S., Steiner D. F. Binding and degradation of 125I-insulin by rat hepatocytes. J Biol Chem. 1975 Nov 10;250(21):8389–8398. [PubMed] [Google Scholar]
  18. Woods J. W., Woodward M. P., Roth T. F. Common features of coated vesicles from dissimilar tissues: composition and structure. J Cell Sci. 1978 Apr;30:87–97. doi: 10.1242/jcs.30.1.87. [DOI] [PubMed] [Google Scholar]

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