Skip to main content
PMC home page
Cell Regulation logoLink to Cell Regulation
. 1991 Aug;2(8):599–612. doi: 10.1091/mbc.2.8.599

Epidermal growth factor and transforming growth factor-alpha: differential intracellular routing and processing of ligand-receptor complexes.

R Ebner 1, R Derynck 1
PMCID: PMC361851  PMID: 1777504

Abstract

Two structurally related but different polypeptide growth factors, epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha), exert their activities after interaction with a common cell-surface EGF/TGF-alpha-receptor. Comparative studies of the effects of both ligands have established that TGF-alpha is more potent than EGF in a variety of biological systems. This observation is not explained by differences in affinities of the ligands for the receptor, because the affinity-constants of both factors are very similar. We have compared the intracellular processing of ligand-receptor complexes using either EGF or TGF-alpha in two different cell systems. We found that TGF-alpha dissociates from the EGF/TGF-alpha-receptor at much higher pH than EGF, which may reflect the substantial difference in the calculated isoelectric points. After internalization, the intracellular TGF-alpha is more rapidly cleared than EGF, and a substantial portion of the released TGF-alpha represents undegraded TGF-alpha in contrast to the mostly degraded EGF. In addition, TGF-alpha did not induce a complete down-regulation of cell surface receptors, as observed with EGF, which is at least in part responsible for a much sooner recovery of the ligand-binding ability after down-regulation, in the case of TGF-alpha. These differences in processing of the ligand-receptor complexes may explain why TGF-alpha exerts quantitatively higher activities than EGF.

Full text

PDF
599

Images in this article

602Image
on p.602
608Image
on p.608

Selected References

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

  1. Anzano M. A., Roberts A. B., Smith J. M., Sporn M. B., De Larco J. E. Sarcoma growth factor from conditioned medium of virally transformed cells is composed of both type alpha and type beta transforming growth factors. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6264–6268. doi: 10.1073/pnas.80.20.6264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barrandon Y., Green H. Cell migration is essential for sustained growth of keratinocyte colonies: the roles of transforming growth factor-alpha and epidermal growth factor. Cell. 1987 Sep 25;50(7):1131–1137. doi: 10.1016/0092-8674(87)90179-6. [DOI] [PubMed] [Google Scholar]
  3. Basu S. K., Goldstein J. L., Anderson R. G., Brown M. S. Monensin interrupts the recycling of low density lipoprotein receptors in human fibroblasts. Cell. 1981 May;24(2):493–502. doi: 10.1016/0092-8674(81)90340-8. [DOI] [PubMed] [Google Scholar]
  4. Bates S. E., Davidson N. E., Valverius E. M., Freter C. E., Dickson R. B., Tam J. P., Kudlow J. E., Lippman M. E., Salomon D. S. Expression of transforming growth factor alpha and its messenger ribonucleic acid in human breast cancer: its regulation by estrogen and its possible functional significance. Mol Endocrinol. 1988 Jun;2(6):543–555. doi: 10.1210/mend-2-6-543. [DOI] [PubMed] [Google Scholar]
  5. Beauchamp R. D., Barnard J. A., McCutchen C. M., Cherner J. A., Coffey R. J., Jr Localization of transforming growth factor alpha and its receptor in gastric mucosal cells. Implications for a regulatory role in acid secretion and mucosal renewal. J Clin Invest. 1989 Sep;84(3):1017–1023. doi: 10.1172/JCI114223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brenner D. A., Koch K. S., Leffert H. L. Transforming growth factor-alpha stimulates proto-oncogene c-jun expression and a mitogenic program in primary cultures of adult rat hepatocytes. DNA. 1989 May;8(4):279–285. doi: 10.1089/dna.1.1989.8.279. [DOI] [PubMed] [Google Scholar]
  7. Brown G. L., Curtsinger L., 3rd, Brightwell J. R., Ackerman D. M., Tobin G. R., Polk H. C., Jr, George-Nascimento C., Valenzuela P., Schultz G. S. Enhancement of epidermal regeneration by biosynthetic epidermal growth factor. J Exp Med. 1986 May 1;163(5):1319–1324. doi: 10.1084/jem.163.5.1319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Carpenter G., Cohen S. Epidermal growth factor. Annu Rev Biochem. 1979;48:193–216. doi: 10.1146/annurev.bi.48.070179.001205. [DOI] [PubMed] [Google Scholar]
  11. Chen W. S., Lazar C. S., Lund K. A., Welsh J. B., Chang C. P., Walton G. M., Der C. J., Wiley H. S., Gill G. N., Rosenfeld M. G. Functional independence of the epidermal growth factor receptor from a domain required for ligand-induced internalization and calcium regulation. Cell. 1989 Oct 6;59(1):33–43. doi: 10.1016/0092-8674(89)90867-2. [DOI] [PubMed] [Google Scholar]
  12. Coffey R. J., Jr, Derynck R., Wilcox J. N., Bringman T. S., Goustin A. S., Moses H. L., Pittelkow M. R. Production and auto-induction of transforming growth factor-alpha in human keratinocytes. 1987 Aug 27-Sep 2Nature. 328(6133):817–820. doi: 10.1038/328817a0. [DOI] [PubMed] [Google Scholar]
  13. Coleman S., Daniel C. W. Inhibition of mouse mammary ductal morphogenesis and down-regulation of the EGF receptor by epidermal growth factor. Dev Biol. 1990 Feb;137(2):425–433. doi: 10.1016/0012-1606(90)90267-m. [DOI] [PubMed] [Google Scholar]
  14. De Larco J. E., Todaro G. J. Sarcoma growth factor (SGF): specific binding to epidermal growth factor (EGF) membrane receptors. J Cell Physiol. 1980 Feb;102(2):267–277. doi: 10.1002/jcp.1041020218. [DOI] [PubMed] [Google Scholar]
  15. Derynck R., Goeddel D. V., Ullrich A., Gutterman J. U., Williams R. D., Bringman T. S., Berger W. H. Synthesis of messenger RNAs for transforming growth factors alpha and beta and the epidermal growth factor receptor by human tumors. Cancer Res. 1987 Feb 1;47(3):707–712. [PubMed] [Google Scholar]
  16. Derynck R. Transforming growth factor alpha. Cell. 1988 Aug 26;54(5):593–595. doi: 10.1016/s0092-8674(88)80001-1. [DOI] [PubMed] [Google Scholar]
  17. Di Fiore P. P., Pierce J. H., Fleming T. P., Hazan R., Ullrich A., King C. R., Schlessinger J., Aaronson S. A. Overexpression of the human EGF receptor confers an EGF-dependent transformed phenotype to NIH 3T3 cells. Cell. 1987 Dec 24;51(6):1063–1070. doi: 10.1016/0092-8674(87)90592-7. [DOI] [PubMed] [Google Scholar]
  18. Di Marco E., Pierce J. H., Fleming T. P., Kraus M. H., Molloy C. J., Aaronson S. A., Di Fiore P. P. Autocrine interaction between TGF alpha and the EGF-receptor: quantitative requirements for induction of the malignant phenotype. Oncogene. 1989 Jul;4(7):831–838. [PubMed] [Google Scholar]
  19. Felder S., Miller K., Moehren G., Ullrich A., Schlessinger J., Hopkins C. R. Kinase activity controls the sorting of the epidermal growth factor receptor within the multivesicular body. Cell. 1990 May 18;61(4):623–634. doi: 10.1016/0092-8674(90)90474-s. [DOI] [PubMed] [Google Scholar]
  20. Frolik C. A., Wakefield L. M., Smith D. M., Sporn M. B. Characterization of a membrane receptor for transforming growth factor-beta in normal rat kidney fibroblasts. J Biol Chem. 1984 Sep 10;259(17):10995–11000. [PubMed] [Google Scholar]
  21. Gan B. S., Hollenberg M. D., MacCannell K. L., Lederis K., Winkler M. E., Derynck R. Distinct vascular actions of epidermal growth factor-urogastrone and transforming growth factor-alpha. J Pharmacol Exp Ther. 1987 Jul;242(1):331–337. [PubMed] [Google Scholar]
  22. Gladhaug I. P., Christoffersen T. Rapid constitutive internalization and externalization of epidermal growth factor receptors in isolated rat hepatocytes. Monensin inhibits receptor externalization and reduces the capacity for continued endocytosis of epidermal growth factor. J Biol Chem. 1988 Sep 5;263(25):12199–12203. [PubMed] [Google Scholar]
  23. Glenney J. R., Jr, Chen W. S., Lazar C. S., Walton G. M., Zokas L. M., Rosenfeld M. G., Gill G. N. Ligand-induced endocytosis of the EGF receptor is blocked by mutational inactivation and by microinjection of anti-phosphotyrosine antibodies. Cell. 1988 Mar 11;52(5):675–684. doi: 10.1016/0092-8674(88)90405-9. [DOI] [PubMed] [Google Scholar]
  24. Gutierrez G. E., Mundy G. R., Derynck R., Hewlett E. L., Katz M. S. Inhibition of parathyroid hormone-responsive adenylate cyclase in clonal osteoblast-like cells by transforming growth factor alpha and epidermal growth factor. J Biol Chem. 1987 Nov 25;262(33):15845–15850. [PubMed] [Google Scholar]
  25. Haigler H. T., Maxfield F. R., Willingham M. C., Pastan I. Dansylcadaverine inhibits internalization of 125I-epidermal growth factor in BALB 3T3 cells. J Biol Chem. 1980 Feb 25;255(4):1239–1241. [PubMed] [Google Scholar]
  26. Han V. K., Hunter E. S., 3rd, Pratt R. M., Zendegui J. G., Lee D. C. Expression of rat transforming growth factor alpha mRNA during development occurs predominantly in the maternal decidua. Mol Cell Biol. 1987 Jul;7(7):2335–2343. doi: 10.1128/mcb.7.7.2335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Higashiyama S., Abraham J. A., Miller J., Fiddes J. C., Klagsbrun M. A heparin-binding growth factor secreted by macrophage-like cells that is related to EGF. Science. 1991 Feb 22;251(4996):936–939. doi: 10.1126/science.1840698. [DOI] [PubMed] [Google Scholar]
  28. Ibbotson K. J., Harrod J., Gowen M., D'Souza S., Smith D. D., Winkler M. E., Derynck R., Mundy G. R. Human recombinant transforming growth factor alpha stimulates bone resorption and inhibits formation in vitro. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2228–2232. doi: 10.1073/pnas.83.7.2228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Jhappan C., Stahle C., Harkins R. N., Fausto N., Smith G. H., Merlino G. T. TGF alpha overexpression in transgenic mice induces liver neoplasia and abnormal development of the mammary gland and pancreas. Cell. 1990 Jun 15;61(6):1137–1146. doi: 10.1016/0092-8674(90)90076-q. [DOI] [PubMed] [Google Scholar]
  30. Kaplan J., Ward D. M., Wiley H. S. Phenylarsine oxide-induced increase in alveolar macrophage surface receptors: evidence for fusion of internal receptor pools with the cell surface. J Cell Biol. 1985 Jul;101(1):121–129. doi: 10.1083/jcb.101.1.121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kawamoto T., Sato J. D., Le A., Polikoff J., Sato G. H., 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. doi: 10.1073/pnas.80.5.1337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Kobrin M. S., Samsoondar J., Kudlow J. E. Alpha-transforming growth factor secreted by untransformed bovine anterior pituitary cells in culture. II. Identification using a sequence-specific monoclonal antibody. J Biol Chem. 1986 Nov 5;261(31):14414–14419. [PubMed] [Google Scholar]
  33. Korc M., Finman J. E. Attenuated processing of epidermal growth factor in the face of marked degradation of transforming growth factor-alpha. J Biol Chem. 1989 Sep 5;264(25):14990–14999. [PubMed] [Google Scholar]
  34. Korc M., Haussler C. A., Trookman N. S. Divergent effects of epidermal growth factor and transforming growth factors on a human endometrial carcinoma cell line. Cancer Res. 1987 Sep 15;47(18):4909–4914. [PubMed] [Google Scholar]
  35. 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]
  36. Lax I., Johnson A., Howk R., Sap J., Bellot F., Winkler M., Ullrich A., Vennstrom B., Schlessinger J., Givol D. Chicken epidermal growth factor (EGF) receptor: cDNA cloning, expression in mouse cells, and differential binding of EGF and transforming growth factor alpha. Mol Cell Biol. 1988 May;8(5):1970–1978. doi: 10.1128/mcb.8.5.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Madtes D. K., Raines E. W., Sakariassen K. S., Assoian R. K., Sporn M. B., Bell G. I., Ross R. Induction of transforming growth factor-alpha in activated human alveolar macrophages. Cell. 1988 Apr 22;53(2):285–293. doi: 10.1016/0092-8674(88)90390-x. [DOI] [PubMed] [Google Scholar]
  38. Massague J., Czech M. P., Iwata K., DeLarco J. E., Todaro G. J. Affinity labeling of a transforming growth factor receptor that does not interact with epidermal growth factor. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6822–6826. doi: 10.1073/pnas.79.22.6822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Massagué J. Epidermal growth factor-like transforming growth factor. II. Interaction with epidermal growth factor receptors in human placenta membranes and A431 cells. J Biol Chem. 1983 Nov 25;258(22):13614–13620. [PubMed] [Google Scholar]
  40. Matsui Y., Halter S. A., Holt J. T., Hogan B. L., Coffey R. J. Development of mammary hyperplasia and neoplasia in MMTV-TGF alpha transgenic mice. Cell. 1990 Jun 15;61(6):1147–1155. doi: 10.1016/0092-8674(90)90077-r. [DOI] [PubMed] [Google Scholar]
  41. Moore G. P., Panaretto B. A., Wallace A. L. Treatment of ewes at different stages of pregnancy with epidermal growth factor: effects on wool growth and plasma concentrations of growth hormone, prolactin, placental lactogen and thyroxine and on foetal development. Acta Endocrinol (Copenh) 1984 Apr;105(4):558–566. doi: 10.1530/acta.0.1050558. [DOI] [PubMed] [Google Scholar]
  42. Myrdal S. E., Twardzik D. R., Auersperg N. Cell-mediated co-action of transforming growth factors: incubation of type beta with normal rat kidney cells produces a soluble activity that prolongs the ruffling response to type alpha. J Cell Biol. 1986 Apr;102(4):1230–1234. doi: 10.1083/jcb.102.4.1230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Münger K., Phelps W. C., Bubb V., Howley P. M., Schlegel R. The E6 and E7 genes of the human papillomavirus type 16 together are necessary and sufficient for transformation of primary human keratinocytes. J Virol. 1989 Oct;63(10):4417–4421. doi: 10.1128/jvi.63.10.4417-4421.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Nistér M., Libermann T. A., Betsholtz C., Pettersson M., Claesson-Welsh L., Heldin C. H., Schlessinger J., Westermark B. Expression of messenger RNAs for platelet-derived growth factor and transforming growth factor-alpha and their receptors in human malignant glioma cell lines. Cancer Res. 1988 Jul 15;48(14):3910–3918. [PubMed] [Google Scholar]
  45. Plowman G. D., Green J. M., McDonald V. L., Neubauer M. G., Disteche C. M., Todaro G. J., Shoyab M. The amphiregulin gene encodes a novel epidermal growth factor-related protein with tumor-inhibitory activity. Mol Cell Biol. 1990 May;10(5):1969–1981. doi: 10.1128/mcb.10.5.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Rall L. B., Scott J., Bell G. I., Crawford R. J., Penschow J. D., Niall H. D., Coghlan J. P. Mouse prepro-epidermal growth factor synthesis by the kidney and other tissues. Nature. 1985 Jan 17;313(5999):228–231. doi: 10.1038/313228a0. [DOI] [PubMed] [Google Scholar]
  47. Rosenthal A., Lindquist P. B., Bringman T. S., Goeddel D. V., Derynck R. Expression in rat fibroblasts of a human transforming growth factor-alpha cDNA results in transformation. Cell. 1986 Jul 18;46(2):301–309. doi: 10.1016/0092-8674(86)90747-6. [DOI] [PubMed] [Google Scholar]
  48. Salomon D. S., Perroteau I., Kidwell W. R., Tam J., Derynck R. Loss of growth responsiveness to epidermal growth factor and enhanced production of alpha-transforming growth factors in ras-transformed mouse mammary epithelial cells. J Cell Physiol. 1987 Mar;130(3):397–409. doi: 10.1002/jcp.1041300313. [DOI] [PubMed] [Google Scholar]
  49. Sandgren E. P., Luetteke N. C., Palmiter R. D., Brinster R. L., Lee D. C. Overexpression of TGF alpha in transgenic mice: induction of epithelial hyperplasia, pancreatic metaplasia, and carcinoma of the breast. Cell. 1990 Jun 15;61(6):1121–1135. doi: 10.1016/0092-8674(90)90075-p. [DOI] [PubMed] [Google Scholar]
  50. Schlessinger J. Allosteric regulation of the epidermal growth factor receptor kinase. J Cell Biol. 1986 Dec;103(6 Pt 1):2067–2072. doi: 10.1083/jcb.103.6.2067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Schreiber A. B., Winkler M. E., Derynck R. Transforming growth factor-alpha: a more potent angiogenic mediator than epidermal growth factor. Science. 1986 Jun 6;232(4755):1250–1253. doi: 10.1126/science.2422759. [DOI] [PubMed] [Google Scholar]
  52. Shoyab M., Plowman G. D., McDonald V. L., Bradley J. G., Todaro G. J. Structure and function of human amphiregulin: a member of the epidermal growth factor family. Science. 1989 Feb 24;243(4894 Pt 1):1074–1076. doi: 10.1126/science.2466334. [DOI] [PubMed] [Google Scholar]
  53. Siegfried J. M. Detection of human lung epithelial cell growth factors produced by a lung carcinoma cell line: use in culture of primary solid lung tumors. Cancer Res. 1987 Jun 1;47(11):2903–2910. [PubMed] [Google Scholar]
  54. Smith J. M., Sporn M. B., Roberts A. B., Derynck R., Winkler M. E., Gregory H. Human transforming growth factor-alpha causes precocious eyelid opening in newborn mice. Nature. 1985 Jun 6;315(6019):515–516. doi: 10.1038/315515a0. [DOI] [PubMed] [Google Scholar]
  55. Stern P. H., Krieger N. S., Nissenson R. A., Williams R. D., Winkler M. E., Derynck R., Strewler G. J. Human transforming growth factor-alpha stimulates bone resorption in vitro. J Clin Invest. 1985 Nov;76(5):2016–2019. doi: 10.1172/JCI112202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Stroobant P., Rice A. P., Gullick W. J., Cheng D. J., Kerr I. M., Waterfield M. D. Purification and characterization of vaccinia virus growth factor. Cell. 1985 Aug;42(1):383–393. doi: 10.1016/s0092-8674(85)80133-1. [DOI] [PubMed] [Google Scholar]
  57. Ullrich A., Schlessinger J. Signal transduction by receptors with tyrosine kinase activity. Cell. 1990 Apr 20;61(2):203–212. doi: 10.1016/0092-8674(90)90801-k. [DOI] [PubMed] [Google Scholar]
  58. Valverius E. M., Bates S. E., Stampfer M. R., Clark R., McCormick F., Salomon D. S., Lippman M. E., Dickson R. B. Transforming growth factor alpha production and epidermal growth factor receptor expression in normal and oncogene transformed human mammary epithelial cells. Mol Endocrinol. 1989 Jan;3(1):203–214. doi: 10.1210/mend-3-1-203. [DOI] [PubMed] [Google Scholar]
  59. Vonderhaar B. K. Local effects of EGF, alpha-TGF, and EGF-like growth factors on lobuloalveolar development of the mouse mammary gland in vivo. J Cell Physiol. 1987 Sep;132(3):581–584. doi: 10.1002/jcp.1041320324. [DOI] [PubMed] [Google Scholar]
  60. Wilcox J. N., Derynck R. Developmental expression of transforming growth factors alpha and beta in mouse fetus. Mol Cell Biol. 1988 Aug;8(8):3415–3422. doi: 10.1128/mcb.8.8.3415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Wilcox J. N., Derynck R. Localization of cells synthesizing transforming growth factor-alpha mRNA in the mouse brain. J Neurosci. 1988 Jun;8(6):1901–1904. doi: 10.1523/JNEUROSCI.08-06-01901.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Wiley H. S., Walsh B. J., Lund K. A. Global modulation of the epidermal growth factor receptor is triggered by occupancy of only a few receptors. Evidence for a binary regulatory system in normal human fibroblasts. J Biol Chem. 1989 Nov 15;264(32):18912–18920. [PubMed] [Google Scholar]
  63. de Larco J. E., Todaro G. J. Growth factors from murine sarcoma virus-transformed cells. Proc Natl Acad Sci U S A. 1978 Aug;75(8):4001–4005. doi: 10.1073/pnas.75.8.4001. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Cell Regulation are provided here courtesy of American Society for Cell Biology

RESOURCES