Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1996 Feb 2;132(4):717–726. doi: 10.1083/jcb.132.4.717

MDC9, a widely expressed cellular disintegrin containing cytoplasmic SH3 ligand domains

PMCID: PMC2199860  PMID: 8647900

Abstract

Cellular disintegrins are a family of proteins that are related to snake venom integrin ligands and metalloproteases. We have cloned and sequenced the mouse and human homologue of a widely expressed cellular disintegrin, which we have termed MDC9 (for metalloprotease/disintegrin/cysteine-rich protein 9). The deduced mouse and human protein sequences are 82% identical. MDC9 contains several distinct protein domains: a signal sequence is followed by a prodomain and a domain with sequence similarity to snake venom metalloproteases, a disintegrin domain, a cysteine-rich region, an EGF repeat, a membrane anchor, and a cytoplasmic tail. The cytoplasmic tail of MDC9 has two proline-rich sequences which can bind the SH3 domain of Src, and may therefore function as SH3 ligand domains. Western blot analysis shows that MDC9 is an approximately 84-kD glycoprotein in all mouse tissues examined, and in NIH 3T3 fibroblast and C2C12 myoblast mouse cell lines. MDC9 can be both cell surface biotinylated and 125I-labeled in NIH 3T3 mouse fibroblasts, indicating that the protein is present on the plasma membrane. Expression of MDC9 in COS-7 cells yields an 84-kD protein, and immunofluorescence analysis of COS-7 cells expressing MDC9 shows a staining pattern that is consistent with a plasma membrane localization. The apparent molecular mass of 84 kD suggests that MDC9 contains a membrane-anchored metalloprotease and disintegrin domain. We propose that MDC9 might function as a membrane-anchored integrin ligand or metalloprotease, or that MDC9 may combine both activities in one protein.

Full Text

The Full Text of this article is available as a PDF (1.9 MB).

Selected References

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

  1. Albelda S. M., Buck C. A. Integrins and other cell adhesion molecules. FASEB J. 1990 Aug;4(11):2868–2880. [PubMed] [Google Scholar]
  2. Alexandropoulos K., Cheng G., Baltimore D. Proline-rich sequences that bind to Src homology 3 domains with individual specificities. Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3110–3114. doi: 10.1073/pnas.92.8.3110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Almeida E. A., Huovila A. P., Sutherland A. E., Stephens L. E., Calarco P. G., Shaw L. M., Mercurio A. M., Sonnenberg A., Primakoff P., Myles D. G. Mouse egg integrin alpha 6 beta 1 functions as a sperm receptor. Cell. 1995 Jun 30;81(7):1095–1104. doi: 10.1016/s0092-8674(05)80014-5. [DOI] [PubMed] [Google Scholar]
  4. Barker H. L., Perry A. C., Jones R., Hall L. Sequence and expression of a monkey testicular transcript encoding tMDC I, a novel member of the metalloproteinase-like, disintegrin-like, cysteine-rich (MDC) protein family. Biochim Biophys Acta. 1994 Aug 2;1218(3):429–431. doi: 10.1016/0167-4781(94)90198-8. [DOI] [PubMed] [Google Scholar]
  5. Birkedal-Hansen H. Proteolytic remodeling of extracellular matrix. Curr Opin Cell Biol. 1995 Oct;7(5):728–735. doi: 10.1016/0955-0674(95)80116-2. [DOI] [PubMed] [Google Scholar]
  6. Blobel C. P., Myles D. G., Primakoff P., White J. M. Proteolytic processing of a protein involved in sperm-egg fusion correlates with acquisition of fertilization competence. J Cell Biol. 1990 Jul;111(1):69–78. doi: 10.1083/jcb.111.1.69. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Blobel C. P., White J. M. Structure, function and evolutionary relationship of proteins containing a disintegrin domain. Curr Opin Cell Biol. 1992 Oct;4(5):760–765. doi: 10.1016/0955-0674(92)90098-w. [DOI] [PubMed] [Google Scholar]
  8. Blobel C. P., Wolfsberg T. G., Turck C. W., Myles D. G., Primakoff P., White J. M. A potential fusion peptide and an integrin ligand domain in a protein active in sperm-egg fusion. Nature. 1992 Mar 19;356(6366):248–252. doi: 10.1038/356248a0. [DOI] [PubMed] [Google Scholar]
  9. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  10. Creemers J. W., Siezen R. J., Roebroek A. J., Ayoubi T. A., Huylebroeck D., Van de Ven W. J. Modulation of furin-mediated proprotein processing activity by site-directed mutagenesis. J Biol Chem. 1993 Oct 15;268(29):21826–21834. [PubMed] [Google Scholar]
  11. Emi M., Katagiri T., Harada Y., Saito H., Inazawa J., Ito I., Kasumi F., Nakamura Y. A novel metalloprotease/disintegrin-like gene at 17q21.3 is somatically rearranged in two primary breast cancers. Nat Genet. 1993 Oct;5(2):151–157. doi: 10.1038/ng1093-151. [DOI] [PubMed] [Google Scholar]
  12. Feng S., Chen J. K., Yu H., Simon J. A., Schreiber S. L. Two binding orientations for peptides to the Src SH3 domain: development of a general model for SH3-ligand interactions. Science. 1994 Nov 18;266(5188):1241–1247. doi: 10.1126/science.7526465. [DOI] [PubMed] [Google Scholar]
  13. Fuller R. S., Brake A. J., Thorner J. Intracellular targeting and structural conservation of a prohormone-processing endoprotease. Science. 1989 Oct 27;246(4929):482–486. doi: 10.1126/science.2683070. [DOI] [PubMed] [Google Scholar]
  14. Gould R. J., Polokoff M. A., Friedman P. A., Huang T. F., Holt J. C., Cook J. J., Niewiarowski S. Disintegrins: a family of integrin inhibitory proteins from viper venoms. Proc Soc Exp Biol Med. 1990 Nov;195(2):168–171. doi: 10.3181/00379727-195-43129b. [DOI] [PubMed] [Google Scholar]
  15. Huang T. F., Holt J. C., Kirby E. P., Niewiarowski S. Trigramin: primary structure and its inhibition of von Willebrand factor binding to glycoprotein IIb/IIIa complex on human platelets. Biochemistry. 1989 Jan 24;28(2):661–666. doi: 10.1021/bi00428a037. [DOI] [PubMed] [Google Scholar]
  16. Huang T. F., Holt J. C., Lukasiewicz H., Niewiarowski S. Trigramin. A low molecular weight peptide inhibiting fibrinogen interaction with platelet receptors expressed on glycoprotein IIb-IIIa complex. J Biol Chem. 1987 Nov 25;262(33):16157–16163. [PubMed] [Google Scholar]
  17. Hynes R. O. Integrins: a family of cell surface receptors. Cell. 1987 Feb 27;48(4):549–554. doi: 10.1016/0092-8674(87)90233-9. [DOI] [PubMed] [Google Scholar]
  18. Hynes R. O. Integrins: versatility, modulation, and signaling in cell adhesion. Cell. 1992 Apr 3;69(1):11–25. doi: 10.1016/0092-8674(92)90115-s. [DOI] [PubMed] [Google Scholar]
  19. Katagiri T., Harada Y., Emi M., Nakamura Y. Human metalloprotease/disintegrin-like (MDC) gene: exon-intron organization and alternative splicing. Cytogenet Cell Genet. 1995;68(1-2):39–44. doi: 10.1159/000133884. [DOI] [PubMed] [Google Scholar]
  20. Kemble G. W., Bodian D. L., Rosé J., Wilson I. A., White J. M. Intermonomer disulfide bonds impair the fusion activity of influenza virus hemagglutinin. J Virol. 1992 Aug;66(8):4940–4950. doi: 10.1128/jvi.66.8.4940-4950.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Krätzschmar J., Haendler B., Langer G., Boidol W., Bringmann P., Alagon A., Donner P., Schleuning W. D. The plasminogen activator family from the salivary gland of the vampire bat Desmodus rotundus: cloning and expression. Gene. 1991 Sep 15;105(2):229–237. doi: 10.1016/0378-1119(91)90155-5. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Lim W. A., Richards F. M., Fox R. O. Structural determinants of peptide-binding orientation and of sequence specificity in SH3 domains. Nature. 1994 Nov 24;372(6504):375–379. doi: 10.1038/372375a0. [DOI] [PubMed] [Google Scholar]
  24. Mayer B. J., Eck M. J. SH3 domains. Minding your p's and q's. Curr Biol. 1995 Apr 1;5(4):364–367. doi: 10.1016/s0960-9822(95)00073-x. [DOI] [PubMed] [Google Scholar]
  25. Miller K. G., Kiehart D. P. Fly division. J Cell Biol. 1995 Oct;131(1):1–5. doi: 10.1083/jcb.131.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Musial J., Niewiarowski S., Rucinski B., Stewart G. J., Cook J. J., Williams J. A., Edmunds L. H., Jr Inhibition of platelet adhesion to surfaces of extracorporeal circuits by disintegrins. RGD-containing peptides from viper venoms. Circulation. 1990 Jul;82(1):261–273. doi: 10.1161/01.cir.82.1.261. [DOI] [PubMed] [Google Scholar]
  27. Myles D. G., Kimmel L. H., Blobel C. P., White J. M., Primakoff P. Identification of a binding site in the disintegrin domain of fertilin required for sperm-egg fusion. Proc Natl Acad Sci U S A. 1994 May 10;91(10):4195–4198. doi: 10.1073/pnas.91.10.4195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pawson T. Protein modules and signalling networks. Nature. 1995 Feb 16;373(6515):573–580. doi: 10.1038/373573a0. [DOI] [PubMed] [Google Scholar]
  29. Pei D., Weiss S. J. Furin-dependent intracellular activation of the human stromelysin-3 zymogen. Nature. 1995 May 18;375(6528):244–247. doi: 10.1038/375244a0. [DOI] [PubMed] [Google Scholar]
  30. Perry A. C., Barker H. L., Jones R., Hall L. Genetic evidence for an additional member of the metalloproteinase-like, disintegrin-like, cysteine-rich (MDC) family of mammalian proteins and its abundant expression in the testis. Biochim Biophys Acta. 1994 Jul 20;1207(1):134–137. doi: 10.1016/0167-4838(94)90062-0. [DOI] [PubMed] [Google Scholar]
  31. Perry A. C., Gichuhi P. M., Jones R., Hall L. Cloning and analysis of monkey fertilin reveals novel alpha subunit isoforms. Biochem J. 1995 May 1;307(Pt 3):843–850. doi: 10.1042/bj3070843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Perry A. C., Jones R., Barker P. J., Hall L. A mammalian epididymal protein with remarkable sequence similarity to snake venom haemorrhagic peptides. Biochem J. 1992 Sep 15;286(Pt 3):671–675. doi: 10.1042/bj2860671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Primakoff P., Hyatt H., Tredick-Kline J. Identification and purification of a sperm surface protein with a potential role in sperm-egg membrane fusion. J Cell Biol. 1987 Jan;104(1):141–149. doi: 10.1083/jcb.104.1.141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Scarborough R. M., Rose J. W., Naughton M. A., Phillips D. R., Nannizzi L., Arfsten A., Campbell A. M., Charo I. F. Characterization of the integrin specificities of disintegrins isolated from American pit viper venoms. J Biol Chem. 1993 Jan 15;268(2):1058–1065. [PubMed] [Google Scholar]
  35. Sparks A. B., Quilliam L. A., Thorn J. M., Der C. J., Kay B. K. Identification and characterization of Src SH3 ligands from phage-displayed random peptide libraries. J Biol Chem. 1994 Sep 30;269(39):23853–23856. [PubMed] [Google Scholar]
  36. Watanabe T., Nakagawa T., Ikemizu J., Nagahama M., Murakami K., Nakayama K. Sequence requirements for precursor cleavage within the constitutive secretory pathway. J Biol Chem. 1992 Apr 25;267(12):8270–8274. [PubMed] [Google Scholar]
  37. Weskamp G., Blobel C. P. A family of cellular proteins related to snake venom disintegrins. Proc Natl Acad Sci U S A. 1994 Mar 29;91(7):2748–2751. doi: 10.1073/pnas.91.7.2748. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Weskamp G., Reichardt L. F. Evidence that biological activity of NGF is mediated through a novel subclass of high affinity receptors. Neuron. 1991 Apr;6(4):649–663. doi: 10.1016/0896-6273(91)90067-a. [DOI] [PubMed] [Google Scholar]
  39. White J. M. Membrane fusion. Science. 1992 Nov 6;258(5084):917–924. doi: 10.1126/science.1439803. [DOI] [PubMed] [Google Scholar]
  40. Wolfsberg T. G., Bazan J. F., Blobel C. P., Myles D. G., Primakoff P., White J. M. The precursor region of a protein active in sperm-egg fusion contains a metalloprotease and a disintegrin domain: structural, functional, and evolutionary implications. Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10783–10787. doi: 10.1073/pnas.90.22.10783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wolfsberg T. G., Straight P. D., Gerena R. L., Huovila A. P., Primakoff P., Myles D. G., White J. M. ADAM, a widely distributed and developmentally regulated gene family encoding membrane proteins with a disintegrin and metalloprotease domain. Dev Biol. 1995 May;169(1):378–383. doi: 10.1006/dbio.1995.1152. [DOI] [PubMed] [Google Scholar]
  42. Yaffe D., Saxel O. Serial passaging and differentiation of myogenic cells isolated from dystrophic mouse muscle. Nature. 1977 Dec 22;270(5639):725–727. doi: 10.1038/270725a0. [DOI] [PubMed] [Google Scholar]
  43. 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]
  44. Yoshida S., Setoguchi M., Higuchi Y., Akizuki S., Yamamoto S. Molecular cloning of cDNA encoding MS2 antigen, a novel cell surface antigen strongly expressed in murine monocytic lineage. Int Immunol. 1990;2(6):585–591. doi: 10.1093/intimm/2.6.585. [DOI] [PubMed] [Google Scholar]
  45. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES