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J Cell Biol. 1989 Dec 1; 109(6): 2731–2739.
PMCID: PMC2115909

Distinct endocytotic pathways in epidermal growth factor-stimulated human carcinoma A431 cells [published erratum appears in J Cell Biol 1990 Mar;110(3):859]


Addition of EGF to human epidermoid carcinoma A431 cells increases the rate of fluid-phase pinocytosis 6-10-fold as measured by horseradish peroxidase uptake (Haigler, H.T., J. A. McKanna, and S. Cohen. 1979. J. Cell Biol. 83:82-90). We show here that in the absence of extracellular Na+ or in the presence of amiloride the stimulation of pinocytosis by EGF is substantially reduced. Amiloride had no effect on the endocytosis of EGF itself or of transferrin, demonstrating that the receptor-mediated endocytotic pathway operated normally under conditions that blocked stimulated pinocytosis. Amiloride blocked EGF- stimulated pinocytosis in both HCO3(-)-containing and HCO3(-)-free media. The EGF-stimulated pinocytotic activity can frequently be localized to areas of the cell where membrane spreading and ruffling are taking place. These results demonstrate that (a) EGF induces a distinct amiloride-sensitive endocytotic pathway on A431 cells; (b) occupied EGF receptors do not utilize this pathway for their own entry; (c) endocytosis of occupied EGF receptors is not in itself sufficient to stimulate pinocytosis.

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Selected References

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  • ABERCROMBIE M, AMBROSE EJ. Interference microscope studies of cell contacts in tissue culture. Exp Cell Res. 1958 Oct;15(2):332–345. [PubMed]
  • Bar-Sagi D, Feramisco JR. Induction of membrane ruffling and fluid-phase pinocytosis in quiescent fibroblasts by ras proteins. Science. 1986 Sep 5;233(4768):1061–1068. [PubMed]
  • Boron WF, De Weer P. Intracellular pH transients in squid giant axons caused by CO2, NH3, and metabolic inhibitors. J Gen Physiol. 1976 Jan;67(1):91–112. [PMC free article] [PubMed]
  • Bretscher MS, Thomson JN. Distribution of ferritin receptors and coated pits on giant HeLa cells. EMBO J. 1983;2(4):599–603. [PMC free article] [PubMed]
  • Brunk U, Schellens J, Westermark B. Influence of epidermal growth factor (EGF) on ruffling activity, pinocytosis and proliferation of cultivated human glia cells. Exp Cell Res. 1976 Dec;103(2):295–302. [PubMed]
  • Cassel D, Whiteley B, Zhuang YX, Glaser L. Mitogen-independent activation of Na+/H+ exchange in human epidermoid carcinoma A431 cells: regulation by medium osmolarity. J Cell Physiol. 1985 Feb;122(2):178–186. [PubMed]
  • Chinkers M, McKanna JA, Cohen S. Rapid induction of morphological changes in human carcinoma cells A-431 by epidermal growth factors. J Cell Biol. 1979 Oct;83(1):260–265. [PMC free article] [PubMed]
  • Connolly JL, Green SA, Greene LA. Comparison of rapid changes in surface morphology and coated pit formation of PC12 cells in response to nerve growth factor, epidermal growth factor, and dibutyryl cyclic AMP. J Cell Biol. 1984 Feb;98(2):457–465. [PMC free article] [PubMed]
  • Cosson P, de Curtis I, Pouysségur J, Griffiths G, Davoust J. Low cytoplasmic pH inhibits endocytosis and transport from the trans-Golgi network to the cell surface. J Cell Biol. 1989 Feb;108(2):377–387. [PMC free article] [PubMed]
  • Davis RJ, Czech MP. Regulation of transferrin receptor expression at the cell surface by insulin-like growth factors, epidermal growth factor and platelet-derived growth factor. EMBO J. 1986 Apr;5(4):653–658. [PMC free article] [PubMed]
  • Davis RJ, Faucher M, Racaniello LK, Carruthers A, Czech MP. Insulin-like growth factor I and epidermal growth factor regulate the expression of transferrin receptors at the cell surface by distinct mechanisms. J Biol Chem. 1987 Sep 25;262(27):13126–13134. [PubMed]
  • Davoust J, Gruenberg J, Howell KE. Two threshold values of low pH block endocytosis at different stages. EMBO J. 1987 Dec 1;6(12):3601–3609. [PMC free article] [PubMed]
  • FAWCETT DW. SURFACE SPECIALIZATIONS OF ABSORBING CELLS. J Histochem Cytochem. 1965 Feb;13:75–91. [PubMed]
  • Fraker PJ, Speck JC., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. [PubMed]
  • Ganz MB, Boyarsky G, Sterzel RB, Boron WF. Arginine vasopressin enhances pHi regulation in the presence of HCO3- by stimulating three acid-base transport systems. Nature. 1989 Feb 16;337(6208):648–651. [PubMed]
  • Goldstein JL, Brown MS, Anderson RG, Russell DW, Schneider WJ. Receptor-mediated endocytosis: concepts emerging from the LDL receptor system. Annu Rev Cell Biol. 1985;1:1–39. [PubMed]
  • Haigler HT, McKanna JA, Cohen S. Direct visualization of the binding and internalization of a ferritin conjugate of epidermal growth factor in human carcinoma cells A-431. J Cell Biol. 1979 May;81(2):382–395. [PMC free article] [PubMed]
  • Haigler HT, McKanna JA, Cohen S. Rapid stimulation of pinocytosis in human carcinoma cells A-431 by epidermal growth factor. J Cell Biol. 1979 Oct;83(1):82–90. [PMC free article] [PubMed]
  • Heuser J. Changes in lysosome shape and distribution correlated with changes in cytoplasmic pH. J Cell Biol. 1989 Mar;108(3):855–864. [PMC free article] [PubMed]
  • Hopkins CR, Trowbridge IS. Internalization and processing of transferrin and the transferrin receptor in human carcinoma A431 cells. J Cell Biol. 1983 Aug;97(2):508–521. [PMC free article] [PubMed]
  • Hopkins CR, Miller K, Beardmore JM. Receptor-mediated endocytosis of transferrin and epidermal growth factor receptors: a comparison of constitutive and ligand-induced uptake. J Cell Sci Suppl. 1985;3:173–186. [PubMed]
  • Huet C, Ash JF, Singer SJ. The antibody-induced clustering and endocytosis of HLA antigens on cultured human fibroblasts. Cell. 1980 Sep;21(2):429–438. [PubMed]
  • L'Allemain G, Franchi A, Cragoe E, Jr, Pouysségur J. Blockade of the Na+/H+ antiport abolishes growth factor-induced DNA synthesis in fibroblasts. Structure-activity relationships in the amiloride series. J Biol Chem. 1984 Apr 10;259(7):4313–4319. [PubMed]
  • Marsh M, Helenius A. Adsorptive endocytosis of Semliki Forest virus. J Mol Biol. 1980 Sep 25;142(3):439–454. [PubMed]
  • Miyata Y, Nishida E, Koyasu S, Yahara I, Sakai H. Regulation by intracellular Ca2+ and cyclic AMP of the growth factor-induced ruffling membrane formation and stimulation of fluid-phase endocytosis and exocytosis. Exp Cell Res. 1989 Apr;181(2):454–462. [PubMed]
  • Montesano R, Roth J, Robert A, Orci L. Non-coated membrane invaginations are involved in binding and internalization of cholera and tetanus toxins. Nature. 1982 Apr 15;296(5858):651–653. [PubMed]
  • Moolenaar WH, Yarden Y, de Laat SW, Schlessinger J. Epidermal growth factor induces electrically silent Na+ influx in human fibroblasts. J Biol Chem. 1982 Jul 25;257(14):8502–8506. [PubMed]
  • Moolenaar WH, Tsien RY, van der Saag PT, de Laat SW. Na+/H+ exchange and cytoplasmic pH in the action of growth factors in human fibroblasts. Nature. 1983 Aug 18;304(5927):645–648. [PubMed]
  • Moolenaar WH, Tertoolen LG, de Laat SW. Phorbol ester and diacylglycerol mimic growth factors in raising cytoplasmic pH. Nature. 1984 Nov 22;312(5992):371–374. [PubMed]
  • Rothenberg P, Glaser L, Schlesinger P, Cassel D. Epidermal growth factor stimulates amiloride-sensitive 22Na+ uptake in A431 cells. Evidence for Na+/H+ exchange. J Biol Chem. 1983 Apr 25;258(8):4883–4889. [PubMed]
  • Rothenberg P, Glaser L, Schlesinger P, Cassel D. Activation of Na+/H+ exchange by epidermal growth factor elevates intracellular pH in A431 cells. J Biol Chem. 1983 Oct 25;258(20):12644–12653. [PubMed]
  • Rozengurt E. Early signals in the mitogenic response. Science. 1986 Oct 10;234(4773):161–166. [PubMed]
  • Sandvig K, Olsnes S, Petersen OW, van Deurs B. Acidification of the cytosol inhibits endocytosis from coated pits. J Cell Biol. 1987 Aug;105(2):679–689. [PMC free article] [PubMed]
  • Schlessinger J. Allosteric regulation of the epidermal growth factor receptor kinase. J Cell Biol. 1986 Dec;103(6 Pt 1):2067–2072. [PMC free article] [PubMed]
  • Smith JB, Rozengurt E. Serum stimulates the Na+,K+ pump in quiescent fibroblasts by increasing Na+ entry. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5560–5564. [PMC free article] [PubMed]
  • Steinman RM, Cohn ZA. The interaction of soluble horseradish peroxidase with mouse peritoneal macrophages in vitro. J Cell Biol. 1972 Oct;55(1):186–204. [PMC free article] [PubMed]
  • Wileman T, Harding C, Stahl P. Receptor-mediated endocytosis. Biochem J. 1985 Nov 15;232(1):1–14. [PMC free article] [PubMed]
  • Wiley HS. Anomalous binding of epidermal growth factor to A431 cells is due to the effect of high receptor densities and a saturable endocytic system. J Cell Biol. 1988 Aug;107(2):801–810. [PMC free article] [PubMed]
  • Zhuang Y, Cragoe EJ, Jr, Shaikewitz T, Glaser L, Cassel D. Characterization of potent Na+/H+ exchange inhibitors from the amiloride series in A431 cells. Biochemistry. 1984 Sep 11;23(19):4481–4488. [PubMed]

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