Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1998 Dec 15;17(24):7260–7272. doi: 10.1093/emboj/17.24.7260

A metalloprotease-disintegrin, MDC9/meltrin-gamma/ADAM9 and PKCdelta are involved in TPA-induced ectodomain shedding of membrane-anchored heparin-binding EGF-like growth factor.

Y Izumi 1, M Hirata 1, H Hasuwa 1, R Iwamoto 1, T Umata 1, K Miyado 1, Y Tamai 1, T Kurisaki 1, A Sehara-Fujisawa 1, S Ohno 1, E Mekada 1
PMCID: PMC1171072  PMID: 9857183

Abstract

The ectodomains of many proteins located at the cell surface are shed upon cell stimulation. One such protein is the heparin-binding EGF-like growth factor (HB-EGF) that exists in a membrane-anchored form which is converted to a soluble form upon cell stimulation with TPA, an activator of protein kinase C (PKC). We show that PKCdelta binds in vivo and in vitro to the cytoplasmic domain of MDC9/meltrin-gamma/ADAM9, a member of the metalloprotease-disintegrin family. Furthermore, the presence of constitutively active PKCdelta or MDC9 results in the shedding of the ectodomain of proHB-EGF, whereas MDC9 mutants lacking the metalloprotease domain, as well as kinase-negative PKCdelta, suppress the TPA-induced shedding of the ectodomain. These results suggest that MDC9 and PKCdelta are involved in the stimulus-coupled shedding of the proHB-EGF ectodomain.

Full Text

The Full Text of this article is available as a PDF (598.5 KB).

Selected References

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

  1. Aicher B., Lerch M. M., Müller T., Schilling J., Ullrich A. Cellular redistribution of protein tyrosine phosphatases LAR and PTPsigma by inducible proteolytic processing. J Cell Biol. 1997 Aug 11;138(3):681–696. doi: 10.1083/jcb.138.3.681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arribas J., Coodly L., Vollmer P., Kishimoto T. K., Rose-John S., Massagué J. Diverse cell surface protein ectodomains are shed by a system sensitive to metalloprotease inhibitors. J Biol Chem. 1996 May 10;271(19):11376–11382. doi: 10.1074/jbc.271.19.11376. [DOI] [PubMed] [Google Scholar]
  3. Arribas J., Massagué J. Transforming growth factor-alpha and beta-amyloid precursor protein share a secretory mechanism. J Cell Biol. 1995 Feb;128(3):433–441. doi: 10.1083/jcb.128.3.433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Black R. A., Rauch C. T., Kozlosky C. J., Peschon J. J., Slack J. L., Wolfson M. F., Castner B. J., Stocking K. L., Reddy P., Srinivasan S. A metalloproteinase disintegrin that releases tumour-necrosis factor-alpha from cells. Nature. 1997 Feb 20;385(6618):729–733. doi: 10.1038/385729a0. [DOI] [PubMed] [Google Scholar]
  5. Blobel C. P. Metalloprotease-disintegrins: links to cell adhesion and cleavage of TNF alpha and Notch. Cell. 1997 Aug 22;90(4):589–592. doi: 10.1016/s0092-8674(00)80519-x. [DOI] [PubMed] [Google Scholar]
  6. Brachmann R., Lindquist P. B., Nagashima M., Kohr W., Lipari T., Napier M., Derynck R. Transmembrane TGF-alpha precursors activate EGF/TGF-alpha receptors. Cell. 1989 Feb 24;56(4):691–700. doi: 10.1016/0092-8674(89)90591-6. [DOI] [PubMed] [Google Scholar]
  7. Chan Y. M., Jan Y. N. Roles for proteolysis and trafficking in notch maturation and signal transduction. Cell. 1998 Aug 21;94(4):423–426. doi: 10.1016/s0092-8674(00)81583-4. [DOI] [PubMed] [Google Scholar]
  8. Davis S., Gale N. W., Aldrich T. H., Maisonpierre P. C., Lhotak V., Pawson T., Goldfarb M., Yancopoulos G. D. Ligands for EPH-related receptor tyrosine kinases that require membrane attachment or clustering for activity. Science. 1994 Nov 4;266(5186):816–819. doi: 10.1126/science.7973638. [DOI] [PubMed] [Google Scholar]
  9. Dethlefsen S. M., Raab G., Moses M. A., Adam R. M., Klagsbrun M., Freeman M. R. Extracellular calcium influx stimulates metalloproteinase cleavage and secretion of heparin-binding EGF-like growth factor independently of protein kinase C. J Cell Biochem. 1998 May 1;69(2):143–153. doi: 10.1002/(sici)1097-4644(19980501)69:2<143::aid-jcb5>3.0.co;2-s. [DOI] [PubMed] [Google Scholar]
  10. Esch F. S., Keim P. S., Beattie E. C., Blacher R. W., Culwell A. R., Oltersdorf T., McClure D., Ward P. J. Cleavage of amyloid beta peptide during constitutive processing of its precursor. Science. 1990 Jun 1;248(4959):1122–1124. doi: 10.1126/science.2111583. [DOI] [PubMed] [Google Scholar]
  11. Feehan C., Darlak K., Kahn J., Walcheck B., Spatola A. F., Kishimoto T. K. Shedding of the lymphocyte L-selectin adhesion molecule is inhibited by a hydroxamic acid-based protease inhibitor. Identification with an L-selectin-alkaline phosphatase reporter. J Biol Chem. 1996 Mar 22;271(12):7019–7024. doi: 10.1074/jbc.271.12.7019. [DOI] [PubMed] [Google Scholar]
  12. Fernandez-Botran R. Soluble cytokine receptors: their role in immunoregulation. FASEB J. 1991 Aug;5(11):2567–2574. doi: 10.1096/fasebj.5.11.1868981. [DOI] [PubMed] [Google Scholar]
  13. Flanagan J. G., Chan D. C., Leder P. Transmembrane form of the kit ligand growth factor is determined by alternative splicing and is missing in the Sld mutant. Cell. 1991 Mar 8;64(5):1025–1035. doi: 10.1016/0092-8674(91)90326-t. [DOI] [PubMed] [Google Scholar]
  14. Fujise A., Mizuno K., Ueda Y., Osada S., Hirai S., Takayanagi A., Shimizu N., Owada M. K., Nakajima H., Ohno S. Specificity of the high affinity interaction of protein kinase C with a physiological substrate, myristoylated alanine-rich protein kinase C substrate. J Biol Chem. 1994 Dec 16;269(50):31642–31648. [PubMed] [Google Scholar]
  15. Goishi K., Higashiyama S., Klagsbrun M., Nakano N., Umata T., Ishikawa M., Mekada E., Taniguchi N. Phorbol ester induces the rapid processing of cell surface heparin-binding EGF-like growth factor: conversion from juxtacrine to paracrine growth factor activity. Mol Biol Cell. 1995 Aug;6(8):967–980. doi: 10.1091/mbc.6.8.967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Golembo M., Raz E., Shilo B. Z. The Drosophila embryonic midline is the site of Spitz processing, and induces activation of the EGF receptor in the ventral ectoderm. Development. 1996 Nov;122(11):3363–3370. doi: 10.1242/dev.122.11.3363. [DOI] [PubMed] [Google Scholar]
  17. Haass C., Selkoe D. J. Cellular processing of beta-amyloid precursor protein and the genesis of amyloid beta-peptide. Cell. 1993 Dec 17;75(6):1039–1042. doi: 10.1016/0092-8674(93)90312-e. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Higashiyama S., Iwamoto R., Goishi K., Raab G., Taniguchi N., Klagsbrun M., Mekada E. The membrane protein CD9/DRAP 27 potentiates the juxtacrine growth factor activity of the membrane-anchored heparin-binding EGF-like growth factor. J Cell Biol. 1995 Mar;128(5):929–938. doi: 10.1083/jcb.128.5.929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hirai S., Izumi Y., Higa K., Kaibuchi K., Mizuno K., Osada S., Suzuki K., Ohno S. Ras-dependent signal transduction is indispensable but not sufficient for the activation of AP1/Jun by PKC delta. EMBO J. 1994 May 15;13(10):2331–2340. doi: 10.1002/j.1460-2075.1994.tb06517.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Howard L., Lu X., Mitchell S., Griffiths S., Glynn P. Molecular cloning of MADM: a catalytically active mammalian disintegrin-metalloprotease expressed in various cell types. Biochem J. 1996 Jul 1;317(Pt 1):45–50. doi: 10.1042/bj3170045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Iwamoto R., Higashiyama S., Mitamura T., Taniguchi N., Klagsbrun M., Mekada E. Heparin-binding EGF-like growth factor, which acts as the diphtheria toxin receptor, forms a complex with membrane protein DRAP27/CD9, which up-regulates functional receptors and diphtheria toxin sensitivity. EMBO J. 1994 May 15;13(10):2322–2330. doi: 10.1002/j.1460-2075.1994.tb06516.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Izumi Y., Hirai S. i., Tamai Y., Fujise-Matsuoka A., Nishimura Y., Ohno S. A protein kinase Cdelta-binding protein SRBC whose expression is induced by serum starvation. J Biol Chem. 1997 Mar 14;272(11):7381–7389. doi: 10.1074/jbc.272.11.7381. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Lanzrein M., Garred O., Olsnes S., Sandvig K. Diphtheria toxin endocytosis and membrane translocation are dependent on the intact membrane-anchored receptor (HB-EGF precursor): studies on the cell-associated receptor cleaved by a metalloprotease in phorbol-ester-treated cells. Biochem J. 1995 Aug 15;310(Pt 1):285–289. doi: 10.1042/bj3100285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Loechel F., Gilpin B. J., Engvall E., Albrechtsen R., Wewer U. M. Human ADAM 12 (meltrin alpha) is an active metalloprotease. J Biol Chem. 1998 Jul 3;273(27):16993–16997. doi: 10.1074/jbc.273.27.16993. [DOI] [PubMed] [Google Scholar]
  27. Massagué J., Pandiella A. Membrane-anchored growth factors. Annu Rev Biochem. 1993;62:515–541. doi: 10.1146/annurev.bi.62.070193.002503. [DOI] [PubMed] [Google Scholar]
  28. Mekada E., Uchida T. Binding properties of diphtheria toxin to cells are altered by mutation in the fragment A domain. J Biol Chem. 1985 Oct 5;260(22):12148–12153. [PubMed] [Google Scholar]
  29. Mitamura T., Higashiyama S., Taniguchi N., Klagsbrun M., Mekada E. Diphtheria toxin binds to the epidermal growth factor (EGF)-like domain of human heparin-binding EGF-like growth factor/diphtheria toxin receptor and inhibits specifically its mitogenic activity. J Biol Chem. 1995 Jan 20;270(3):1015–1019. doi: 10.1074/jbc.270.3.1015. [DOI] [PubMed] [Google Scholar]
  30. Mitamura T., Iwamoto R., Umata T., Yomo T., Urabe I., Tsuneoka M., Mekada E. The 27-kD diphtheria toxin receptor-associated protein (DRAP27) from vero cells is the monkey homologue of human CD9 antigen: expression of DRAP27 elevates the number of diphtheria toxin receptors on toxin-sensitive cells. J Cell Biol. 1992 Sep;118(6):1389–1399. doi: 10.1083/jcb.118.6.1389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Miyoshi E., Higashiyama S., Nakagawa T., Hayashi N., Taniguchi N. Membrane-anchored heparin-binding epidermal growth factor-like growth factor acts as a tumor survival factor in a hepatoma cell line. J Biol Chem. 1997 May 30;272(22):14349–14355. doi: 10.1074/jbc.272.22.14349. [DOI] [PubMed] [Google Scholar]
  32. Mizuno K., Kubo K., Saido T. C., Akita Y., Osada S., Kuroki T., Ohno S., Suzuki K. Structure and properties of a ubiquitously expressed protein kinase C, nPKC delta. Eur J Biochem. 1991 Dec 18;202(3):931–940. doi: 10.1111/j.1432-1033.1991.tb16453.x. [DOI] [PubMed] [Google Scholar]
  33. Mizushima S., Nagata S. pEF-BOS, a powerful mammalian expression vector. Nucleic Acids Res. 1990 Sep 11;18(17):5322–5322. doi: 10.1093/nar/18.17.5322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Mohler K. M., Sleath P. R., Fitzner J. N., Cerretti D. P., Alderson M., Kerwar S. S., Torrance D. S., Otten-Evans C., Greenstreet T., Weerawarna K. Protection against a lethal dose of endotoxin by an inhibitor of tumour necrosis factor processing. Nature. 1994 Jul 21;370(6486):218–220. doi: 10.1038/370218a0. [DOI] [PubMed] [Google Scholar]
  35. Moss M. L., Jin S. L., Milla M. E., Bickett D. M., Burkhart W., Carter H. L., Chen W. J., Clay W. C., Didsbury J. R., Hassler D. Cloning of a disintegrin metalloproteinase that processes precursor tumour-necrosis factor-alpha. Nature. 1997 Feb 20;385(6618):733–736. doi: 10.1038/385733a0. [DOI] [PubMed] [Google Scholar]
  36. Naglich J. G., Metherall J. E., Russell D. W., Eidels L. Expression cloning of a diphtheria toxin receptor: identity with a heparin-binding EGF-like growth factor precursor. Cell. 1992 Jun 12;69(6):1051–1061. doi: 10.1016/0092-8674(92)90623-k. [DOI] [PubMed] [Google Scholar]
  37. Nakagawa T., Higashiyama S., Mitamura T., Mekada E., Taniguchi N. Amino-terminal processing of cell surface heparin-binding epidermal growth factor-like growth factor up-regulates its juxtacrine but not its paracrine growth factor activity. J Biol Chem. 1996 Nov 29;271(48):30858–30863. doi: 10.1074/jbc.271.48.30858. [DOI] [PubMed] [Google Scholar]
  38. Nakamura K., Iwamoto R., Mekada E. Membrane-anchored heparin-binding EGF-like growth factor (HB-EGF) and diphtheria toxin receptor-associated protein (DRAP27)/CD9 form a complex with integrin alpha 3 beta 1 at cell-cell contact sites. J Cell Biol. 1995 Jun;129(6):1691–1705. doi: 10.1083/jcb.129.6.1691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Ohno S., Mizuno K., Adachi Y., Hata A., Akita Y., Akimoto K., Osada S., Hirai S., Suzuki K. Activation of novel protein kinases C delta and C epsilon upon mitogenic stimulation of quiescent rat 3Y1 fibroblasts. J Biol Chem. 1994 Jul 1;269(26):17495–17501. [PubMed] [Google Scholar]
  40. Pan D., Rubin G. M. Kuzbanian controls proteolytic processing of Notch and mediates lateral inhibition during Drosophila and vertebrate neurogenesis. Cell. 1997 Jul 25;90(2):271–280. doi: 10.1016/s0092-8674(00)80335-9. [DOI] [PubMed] [Google Scholar]
  41. Pandiella A., Massagué J. Cleavage of the membrane precursor for transforming growth factor alpha is a regulated process. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1726–1730. doi: 10.1073/pnas.88.5.1726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Pandiella A., Massagué J. Multiple signals activate cleavage of the membrane transforming growth factor-alpha precursor. J Biol Chem. 1991 Mar 25;266(9):5769–5773. [PubMed] [Google Scholar]
  43. Pierce J. H., Ruggiero M., Fleming T. P., Di Fiore P. P., Greenberger J. S., Varticovski L., Schlessinger J., Rovera G., Aaronson S. A. Signal transduction through the EGF receptor transfected in IL-3-dependent hematopoietic cells. Science. 1988 Feb 5;239(4840):628–631. doi: 10.1126/science.3257584. [DOI] [PubMed] [Google Scholar]
  44. Rose-John S., Heinrich P. C. Soluble receptors for cytokines and growth factors: generation and biological function. Biochem J. 1994 Jun 1;300(Pt 2):281–290. doi: 10.1042/bj3000281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Shing Y., Christofori G., Hanahan D., Ono Y., Sasada R., Igarashi K., Folkman J. Betacellulin: a mitogen from pancreatic beta cell tumors. Science. 1993 Mar 12;259(5101):1604–1607. doi: 10.1126/science.8456283. [DOI] [PubMed] [Google Scholar]
  46. 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]
  47. Sisodia S. S., Price D. L. Role of the beta-amyloid protein in Alzheimer's disease. FASEB J. 1995 Mar;9(5):366–370. doi: 10.1096/fasebj.9.5.7896005. [DOI] [PubMed] [Google Scholar]
  48. Stein J., Borzillo G. V., Rettenmier C. W. Direct stimulation of cells expressing receptors for macrophage colony-stimulating factor (CSF-1) by a plasma membrane-bound precursor of human CSF-1. Blood. 1990 Oct 1;76(7):1308–1314. [PubMed] [Google Scholar]
  49. Subramanian S. V., Fitzgerald M. L., Bernfield M. Regulated shedding of syndecan-1 and -4 ectodomains by thrombin and growth factor receptor activation. J Biol Chem. 1997 Jun 6;272(23):14713–14720. doi: 10.1074/jbc.272.23.14713. [DOI] [PubMed] [Google Scholar]
  50. Suzuki M., Raab G., Moses M. A., Fernandez C. A., Klagsbrun M. Matrix metalloproteinase-3 releases active heparin-binding EGF-like growth factor by cleavage at a specific juxtamembrane site. J Biol Chem. 1997 Dec 12;272(50):31730–31737. doi: 10.1074/jbc.272.50.31730. [DOI] [PubMed] [Google Scholar]
  51. Takemura T., Kondo S., Homma T., Sakai M., Harris R. C. The membrane-bound form of heparin-binding epidermal growth factor-like growth factor promotes survival of cultured renal epithelial cells. J Biol Chem. 1997 Dec 5;272(49):31036–31042. doi: 10.1074/jbc.272.49.31036. [DOI] [PubMed] [Google Scholar]
  52. Tanaka M., Itai T., Adachi M., Nagata S. Downregulation of Fas ligand by shedding. Nat Med. 1998 Jan;4(1):31–36. doi: 10.1038/nm0198-031. [DOI] [PubMed] [Google Scholar]
  53. Toyoda H., Komurasaki T., Uchida D., Takayama Y., Isobe T., Okuyama T., Hanada K. Epiregulin. A novel epidermal growth factor with mitogenic activity for rat primary hepatocytes. J Biol Chem. 1995 Mar 31;270(13):7495–7500. doi: 10.1074/jbc.270.13.7495. [DOI] [PubMed] [Google Scholar]
  54. Tsuneoka M., Nakayama K., Hatsuzawa K., Komada M., Kitamura N., Mekada E. Evidence for involvement of furin in cleavage and activation of diphtheria toxin. J Biol Chem. 1993 Dec 15;268(35):26461–26465. [PubMed] [Google Scholar]
  55. Uchida T., Pappenheimer A. M., Jr, Greany R. Diphtheria toxin and related proteins. I. Isolation and properties of mutant proteins serologically related to diphtheria toxin. J Biol Chem. 1973 Jun 10;248(11):3838–3844. [PubMed] [Google Scholar]
  56. Ueda Y., Hirai S. i., Osada S. i., Suzuki A., Mizuno K., Ohno S. Protein kinase C activates the MEK-ERK pathway in a manner independent of Ras and dependent on Raf. J Biol Chem. 1996 Sep 20;271(38):23512–23519. doi: 10.1074/jbc.271.38.23512. [DOI] [PubMed] [Google Scholar]
  57. Vecchi M., Baulida J., Carpenter G. Selective cleavage of the heregulin receptor ErbB-4 by protein kinase C activation. J Biol Chem. 1996 Aug 2;271(31):18989–18995. doi: 10.1074/jbc.271.31.18989. [DOI] [PubMed] [Google Scholar]
  58. Werb Z. ECM and cell surface proteolysis: regulating cellular ecology. Cell. 1997 Nov 14;91(4):439–442. doi: 10.1016/s0092-8674(00)80429-8. [DOI] [PubMed] [Google Scholar]
  59. Weskamp G., Krätzschmar J., Reid M. S., Blobel C. P. MDC9, a widely expressed cellular disintegrin containing cytoplasmic SH3 ligand domains. J Cell Biol. 1996 Feb;132(4):717–726. doi: 10.1083/jcb.132.4.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Wolfsberg T. G., Primakoff P., Myles D. G., White J. M. ADAM, a novel family of membrane proteins containing A Disintegrin And Metalloprotease domain: multipotential functions in cell-cell and cell-matrix interactions. J Cell Biol. 1995 Oct;131(2):275–278. doi: 10.1083/jcb.131.2.275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Wong S. T., Winchell L. F., McCune B. K., Earp H. S., Teixidó J., Massagué J., Herman B., Lee D. C. The TGF-alpha precursor expressed on the cell surface binds to the EGF receptor on adjacent cells, leading to signal transduction. Cell. 1989 Feb 10;56(3):495–506. doi: 10.1016/0092-8674(89)90252-3. [DOI] [PubMed] [Google Scholar]
  62. Yagami-Hiromasa T., Sato T., Kurisaki T., Kamijo K., Nabeshima Y., Fujisawa-Sehara A. A metalloprotease-disintegrin participating in myoblast fusion. Nature. 1995 Oct 19;377(6550):652–656. doi: 10.1038/377652a0. [DOI] [PubMed] [Google Scholar]
  63. Yamamoto M., Tsujishita H., Hori N., Ohishi Y., Inoue S., Ikeda S., Okada Y. Inhibition of membrane-type 1 matrix metalloproteinase by hydroxamate inhibitors: an examination of the subsite pocket. J Med Chem. 1998 Apr 9;41(8):1209–1217. doi: 10.1021/jm970404a. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES