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EMBO J. Aug 1984; 3(8): 1843–1847.
PMCID: PMC557607

High affinity epidermal growth factor receptors on the surface of A431 cells have restricted lateral diffusion.


Rhodamine-labelled epidermal growth factor (Rh-EGF) was shown to bind to A431 cells grown at low density both to a small number of high affinity receptors (KD = 2.8 X 10(-10) M; fraction of total binding sites approximately 0.12) and also to a large number of low affinity receptors (KD = 4 X 10(-9) M; fraction of total binding sites approximately 0.88). Measurements of the lateral diffusion of EGF receptors on the cell surface were made using Rh-EGF and the technique of fluorescence photobleaching recovery. The high affinity receptors (labelled with 1.6 X 10(-10) M Rh-EGF, 5% of EGF binding sites occupied) did not show lateral mobility over the temperature range 3 degrees-37 degrees C. The low affinity receptors (labelled with 2.4 X 10(-7) M Rh-EGF, 90% of EGF sites occupied) showed at least 75% fluorescence recovery after photobleaching, and lateral diffusion coefficients of approximately 2 X 10(-10) cm2/s. These results show that the two populations of EGF receptors defined by binding studies differ in their freedom to diffuse laterally. The observation that the high affinity receptors are immobile indicates that lateral diffusion of receptors, at least over a distance of a few hundred nanometres or more, may not be required for the action of low concentrations of EGF.

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Selected References

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  • King AC, Cuatrecasas P. Resolution of high and low affinity epidermal growth factor receptors. Inhibition of high affinity component by low temperature, cycloheximide, and phorbol esters. J Biol Chem. 1982 Mar 25;257(6):3053–3060. [PubMed]
  • Krupp MN, Connolly DT, Lane MD. Synthesis, turnover, and down-regulation of epidermal growth factor receptors in human A431 epidermoid carcinoma cells and skin fibroblasts. J Biol Chem. 1982 Oct 10;257(19):11489–11496. [PubMed]
  • Lee LS. Saccharin and cyclamate inhibit binding of epidermal growth factor. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1042–1046. [PMC free article] [PubMed]
  • Lifshitz A, Lazar CS, Buss JE, Gill GN. Analysis of morphology and receptor metabolism in clonal variant A431 cells with differing growth responses to epidermal growth factor. J Cell Physiol. 1983 Jun;115(3):235–242. [PubMed]
  • Magun BE, Matrisian LM, Bowden GT. Epidermal growth factor. Ability of tumor promoter to alter its degradation, receptor affinity and receptor number. J Biol Chem. 1980 Jul 10;255(13):6373–6381. [PubMed]
  • Osborene CK, Hamilton B, Nover M. Receptor binding and processing of epidermal growth factor by human breast cancer cells. J Clin Endocrinol Metab. 1982 Jul;55(1):86–93. [PubMed]
  • Rees AR, Offord RE. The preparation of protected fragments of lysozyme for semisynthesis. Biochem J. 1976 Dec 1;159(3):467–479. [PMC free article] [PubMed]
  • Rees AR, Adamson ED, Graham CF. Epidermal growth factor receptors increase during the differentiation of embryonal carcinoma cells. Nature. 1979 Sep 27;281(5729):309–311. [PubMed]
  • Rose SP, Pruss RM, Herschman HR. Initiation of 3T3 fibroblast cell division by epidermal growth factor. J Cell Physiol. 1975 Dec;86 (Suppl 2)(3 Pt 2):593–598. [PubMed]
  • Rozengurt E, Collins M, Brown KD, Pettican P. Inhibition of epidermal growth factor binding to mouse cultured cells by fibroblast-derived growth factor. Evidence for an indirect mechanism. J Biol Chem. 1982 Apr 10;257(7):3680–3686. [PubMed]
  • Saito M, Ueno I, Egawa K. Effect of retinoic acid and 12-O-tetradecanoyl phorbol-13-acetate on the binding of epidermal growth factor to its cellular receptors. Biochim Biophys Acta. 1982 Aug 6;717(2):301–304. [PubMed]
  • Schlessinger J, Shechter Y, Willingham MC, Pastan I. Direct visualization of binding, aggregation, and internalization of insulin and epidermal growth factor on living fibroblastic cells. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2659–2663. [PMC free article] [PubMed]
  • Schlessinger J, Shechter Y, Cuatrecasas P, Willingham MC, Pastan I. Quantitative determination of the lateral diffusion coefficients of the hormone-receptor complexes of insulin and epidermal growth factor on the plasma membrane of cultured fibroblasts. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5353–5357. [PMC free article] [PubMed]
  • Shechter Y, Hernaez L, Cuatrecasas P. Epidermal growth factor: biological activity requires persistent occupation of high-affinity cell surface receptors. Proc Natl Acad Sci U S A. 1978 Dec;75(12):5788–5791. [PMC free article] [PubMed]
  • Shoyab M, De Larco JE, Todaro GJ. Biologically active phorbol esters specifically alter affinity of epidermal growth factor membrane receptors. Nature. 1979 May 31;279(5712):387–391. [PubMed]
  • Todaro GJ, De Larco JE, Cohen S. Transformation by murine and feline sarcoma viruses specifically blocks binding of epidermal growth factor to cells. Nature. 1976 Nov 4;264(5581):26–31. [PubMed]
  • Willingham MC, Haigler HT, Fitzgerald DJ, Gallo MG, Rutherford AV, Pastan IH. The morphologic pathway of binding and internalization of epidermal growth factor in cultured cells. Studies on A431, KB, and 3T3 cells, using multiple methods of labelling. Exp Cell Res. 1983 Jun;146(1):163–175. [PubMed]
  • Zidovetzki R, Yarden Y, Schlessinger J, Jovin TM. Rotational diffusion of epidermal growth factor complexed to cell surface receptors reflects rapid microaggregation and endocytosis of occupied receptors. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6981–6985. [PMC free article] [PubMed]
  • Adamson ED, Rees AR. Epidermal growth factor receptors. Mol Cell Biochem. 1981 Feb 11;34(3):129–152. [PubMed]
  • Aharonov A, Pruss RM, Herschman HR. Epidermal growth factor. Relationship between receptor regulation and mitogenesis in 3T3 cells. J Biol Chem. 1978 Jun 10;253(11):3970–3977. [PubMed]
  • Axelrod D, Koppel DE, Schlessinger J, Elson E, Webb WW. Mobility measurement by analysis of fluorescence photobleaching recovery kinetics. Biophys J. 1976 Sep;16(9):1055–1069. [PMC free article] [PubMed]
  • Barnes DW. Epidermal growth factor inhibits growth of A431 human epidermoid carcinoma in serum-free cell culture. J Cell Biol. 1982 Apr;93(1):1–4. [PMC free article] [PubMed]
  • Carpenter G, Cohen S. Human epidermal growth factor and the proliferation of human fibroblasts. J Cell Physiol. 1976 Jun;88(2):227–237. [PubMed]
  • Collins MK, Sinnett-Smith JW, Rozengurt E. Platelet-derived growth factor treatment decreases the affinity of the epidermal growth factor receptors of Swiss 3T3 cells. J Biol Chem. 1983 Oct 10;258(19):11689–11693. [PubMed]
  • Das M, Fox CF. Molecular mechanism of mitogen action: processing of receptor induced by epidermal growth factor. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2644–2648. [PMC free article] [PubMed]
  • Fabricant RN, De Larco JE, Todaro GJ. Nerve growth factor receptors on human melanoma cells in culture. Proc Natl Acad Sci U S A. 1977 Feb;74(2):565–569. [PMC free article] [PubMed]
  • Fox CF, Das M. Internalization and processing of the EGF receptor in the induction of DNA synthesis in cultured fibroblasts: the endocytic activation hypothesis. J Supramol Struct. 1979;10(2):199–214. [PubMed]
  • Garland P. Fluorescence photobleaching recovery: control of laser intensities with an acousto-optic modulator. Biophys J. 1981 Mar;33(3):481–482. [PMC free article] [PubMed]
  • Gill GN, Lazar CS. Increased phosphotyrosine content and inhibition of proliferation in EGF-treated A431 cells. Nature. 1981 Sep 24;293(5830):305–307. [PubMed]
  • Gregorou M, Rees AR. Properties of a monoclonal antibody to epidermal growth factor receptor with implications for the mechanism of action of EGF. EMBO J. 1984 May;3(5):929–937. [PMC free article] [PubMed]
  • Haigler H, Ash JF, Singer SJ, Cohen S. Visualization by fluorescence of the binding and internalization of epidermal growth factor in human carcinoma cells A-431. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3317–3321. [PMC free article] [PubMed]
  • Hillman GM, Schlessinger J. Lateral diffusion of epidermal growth factor complexed to its surface receptors does not account for the thermal sensitivity of patch formation and endocytosis. Biochemistry. 1982 Mar 30;21(7):1667–1672. [PubMed]
  • Johnson P, Garland PB. Depolarization of fluorescence depletion. A microscopic method for measuring rotational diffusion of membrane proteins on the surface of a single cell. FEBS Lett. 1981 Sep 28;132(2):252–256. [PubMed]
  • Kawamoto T, Sato JD, Le A, Polikoff J, Sato GH, Mendelsohn J. Growth stimulation of A431 cells by epidermal growth factor: identification of high-affinity receptors for epidermal growth factor by an anti-receptor monoclonal antibody. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1337–1341. [PMC free article] [PubMed]

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