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. 1996 Apr 2;133(2):359–369. doi: 10.1083/jcb.133.2.359

The binding of plakoglobin to desmosomal cadherins: patterns of binding sites and topogenic potential

PMCID: PMC2120792  PMID: 8609168

Abstract

Plakoglobin is the only protein that occurs in the cytoplasmic plaques of all known adhering junctions and has been shown to be crucially involved in the formation and maintenance of desmosomes anchoring intermediate-sized filaments (IFs) by its interaction with the desmosomal cadherins, desmoglein (Dsg), and desmocollin (Dsc). This topogenic importance of plakoglobin is now directly shown in living cells as well as in binding assays in vitro. We show that, in transfected human A-431 carcinoma cells, a chimeric protein combining the vesicle-forming transmembrane glycoprotein synaptophysin, with the complete human plakoglobin sequence, is sorted to small vesicles many of which associate with desmosomal plaques and their attached IFs. Immunoprecipitation experiments have further revealed that the chimeric plakoglobin-containing transmembrane molecules of these vesicles are tightly bound to Dsg and Dsc but not to endogenous plakoglobin, thus demonstrating that the binding of plakoglobin to desmosomal cadherins does not require its soluble state and is strong enough to attach large structures such as vesicles to desmosomes. To identify the binding domains and the mechanisms involved in the interaction of plakoglobin with desmosomal cadherins, we have developed direct binding assays in vitro in which plakoglobin or parts thereof, produced by recombinant DNA technology in E. coli, are exposed to molecules containing the "C- domains" of several cadherins. These assays have shown that plakoglobin associates most tightly with the C-domain of Dsg, to a lesser degree with that of Dsc and only weakly with the C-domain of E-cadherin. Three separate segments of plakoglobin containing various numbers of the so- called arm repeats exhibit distinct binding to the desmosomal cadherins comparable in strength to that of the entire molecule. The binding pattern of plakoglobin segments in vitro is compared with that in vivo. Paradoxically, in vitro some internal plakoglobin fragments bind even better to the C-domain of E-cadherin than the entire molecule, indicating that elements exist in native plakoglobin that interfere with the interaction of this protein with its various cadherin partners.

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Selected References

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  1. Aberle H., Butz S., Stappert J., Weissig H., Kemler R., Hoschuetzky H. Assembly of the cadherin-catenin complex in vitro with recombinant proteins. J Cell Sci. 1994 Dec;107(Pt 12):3655–3663. doi: 10.1242/jcs.107.12.3655. [DOI] [PubMed] [Google Scholar]
  2. Butz S., Stappert J., Weissig H., Kemler R. Plakoglobin and beta-catenin: distinct but closely related. Science. 1992 Aug 21;257(5073):1142–1144. doi: 10.1126/science.257.5073.1142-a. [DOI] [PubMed] [Google Scholar]
  3. Buxton R. S., Cowin P., Franke W. W., Garrod D. R., Green K. J., King I. A., Koch P. J., Magee A. I., Rees D. A., Stanley J. R. Nomenclature of the desmosomal cadherins. J Cell Biol. 1993 May;121(3):481–483. doi: 10.1083/jcb.121.3.481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cameron P. L., Südhof T. C., Jahn R., De Camilli P. Colocalization of synaptophysin with transferrin receptors: implications for synaptic vesicle biogenesis. J Cell Biol. 1991 Oct;115(1):151–164. doi: 10.1083/jcb.115.1.151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cowin P., Kapprell H. P., Franke W. W., Tamkun J., Hynes R. O. Plakoglobin: a protein common to different kinds of intercellular adhering junctions. Cell. 1986 Sep 26;46(7):1063–1073. doi: 10.1016/0092-8674(86)90706-3. [DOI] [PubMed] [Google Scholar]
  6. Cowin P., Kapprell H. P., Franke W. W. The complement of desmosomal plaque proteins in different cell types. J Cell Biol. 1985 Oct;101(4):1442–1454. doi: 10.1083/jcb.101.4.1442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Demlehner M. P., Schäfer S., Grund C., Franke W. W. Continual assembly of half-desmosomal structures in the absence of cell contacts and their frustrated endocytosis: a coordinated Sisyphus cycle. J Cell Biol. 1995 Nov;131(3):745–760. doi: 10.1083/jcb.131.3.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fouquet B., Zimbelmann R., Franke W. W. Identification of plakoglobin in oocytes and early embryos of Xenopus laevis: maternal expression of a gene encoding a junctional plaque protein. Differentiation. 1992 Nov;51(3):187–194. doi: 10.1111/j.1432-0436.1992.tb00695.x. [DOI] [PubMed] [Google Scholar]
  9. Franke W. W., Goldschmidt M. D., Zimbelmann R., Mueller H. M., Schiller D. L., Cowin P. Molecular cloning and amino acid sequence of human plakoglobin, the common junctional plaque protein. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4027–4031. doi: 10.1073/pnas.86.11.4027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Franke W. W., Kapprell H. P., Cowin P. Immunolocalization of plakoglobin in endothelial junctions: identification as a special type of Zonulae adhaerentes. Biol Cell. 1987;59(3):205–218. doi: 10.1111/j.1768-322x.1987.tb00532.x. [DOI] [PubMed] [Google Scholar]
  11. Franke W. W., Kapprell H. P., Cowin P. Plakoglobin is a component of the filamentous subplasmalemmal coat of lens cells. Eur J Cell Biol. 1987 Jun;43(3):301–315. [PubMed] [Google Scholar]
  12. Franke W. W., Troyanovsky S. M., Koch P. J., Troyanovsky R., Fouquet B., Leube R. E. Desmosomal proteins: mediators of intercellular coupling and intermediate filament anchorage. Cold Spring Harb Symp Quant Biol. 1992;57:643–652. doi: 10.1101/sqb.1992.057.01.070. [DOI] [PubMed] [Google Scholar]
  13. Franke W. W., Winter S., von Overbeck J., Gudat F., Heitz P. U., Stähli C. Identification of the conserved, conformation-dependent cytokeratin epitope recognized by monoclonal antibody (lu-5). Virchows Arch A Pathol Anat Histopathol. 1987;411(2):137–147. doi: 10.1007/BF00712737. [DOI] [PubMed] [Google Scholar]
  14. Garrod D. R. Desmosomes and hemidesmosomes. Curr Opin Cell Biol. 1993 Feb;5(1):30–40. doi: 10.1016/s0955-0674(05)80005-5. [DOI] [PubMed] [Google Scholar]
  15. Grunwald G. B. The structural and functional analysis of cadherin calcium-dependent cell adhesion molecules. Curr Opin Cell Biol. 1993 Oct;5(5):797–805. doi: 10.1016/0955-0674(93)90028-o. [DOI] [PubMed] [Google Scholar]
  16. Hatzfeld M., Kristjansson G. I., Plessmann U., Weber K. Band 6 protein, a major constituent of desmosomes from stratified epithelia, is a novel member of the armadillo multigene family. J Cell Sci. 1994 Aug;107(Pt 8):2259–2270. doi: 10.1242/jcs.107.8.2259. [DOI] [PubMed] [Google Scholar]
  17. Heid H. W., Schmidt A., Zimbelmann R., Schäfer S., Winter-Simanowski S., Stumpp S., Keith M., Figge U., Schnölzer M., Franke W. W. Cell type-specific desmosomal plaque proteins of the plakoglobin family: plakophilin 1 (band 6 protein). Differentiation. 1994 Dec;58(2):113–131. doi: 10.1046/j.1432-0436.1995.5820113.x. [DOI] [PubMed] [Google Scholar]
  18. Hinck L., Näthke I. S., Papkoff J., Nelson W. J. Dynamics of cadherin/catenin complex formation: novel protein interactions and pathways of complex assembly. J Cell Biol. 1994 Jun;125(6):1327–1340. doi: 10.1083/jcb.125.6.1327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Horst M., Harth N., Hasilik A. Biosynthesis of glycosylated human lysozyme mutants. J Biol Chem. 1991 Jul 25;266(21):13914–13919. [PubMed] [Google Scholar]
  20. Hoschuetzky H., Aberle H., Kemler R. Beta-catenin mediates the interaction of the cadherin-catenin complex with epidermal growth factor receptor. J Cell Biol. 1994 Dec;127(5):1375–1380. doi: 10.1083/jcb.127.5.1375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hülsken J., Birchmeier W., Behrens J. E-cadherin and APC compete for the interaction with beta-catenin and the cytoskeleton. J Cell Biol. 1994 Dec;127(6 Pt 2):2061–2069. doi: 10.1083/jcb.127.6.2061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jou T. S., Stewart D. B., Stappert J., Nelson W. J., Marrs J. A. Genetic and biochemical dissection of protein linkages in the cadherin-catenin complex. Proc Natl Acad Sci U S A. 1995 May 23;92(11):5067–5071. doi: 10.1073/pnas.92.11.5067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kapprell H. P., Cowin P., Franke W. W. Biochemical characterization of the soluble form of the junctional plaque protein, plakoglobin, from different cell types. Eur J Biochem. 1987 Aug 3;166(3):505–517. doi: 10.1111/j.1432-1033.1987.tb13543.x. [DOI] [PubMed] [Google Scholar]
  24. Kemler R. From cadherins to catenins: cytoplasmic protein interactions and regulation of cell adhesion. Trends Genet. 1993 Sep;9(9):317–321. doi: 10.1016/0168-9525(93)90250-l. [DOI] [PubMed] [Google Scholar]
  25. Knudsen K. A., Wheelock M. J. Plakoglobin, or an 83-kD homologue distinct from beta-catenin, interacts with E-cadherin and N-cadherin. J Cell Biol. 1992 Aug;118(3):671–679. doi: 10.1083/jcb.118.3.671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Koch P. J., Franke W. W. Desmosomal cadherins: another growing multigene family of adhesion molecules. Curr Opin Cell Biol. 1994 Oct;6(5):682–687. doi: 10.1016/0955-0674(94)90094-9. [DOI] [PubMed] [Google Scholar]
  27. Koch P. J., Goldschmidt M. D., Zimbelmann R., Troyanovsky R., Franke W. W. Complexity and expression patterns of the desmosomal cadherins. Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):353–357. doi: 10.1073/pnas.89.1.353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Koch P. J., Walsh M. J., Schmelz M., Goldschmidt M. D., Zimbelmann R., Franke W. W. Identification of desmoglein, a constitutive desmosomal glycoprotein, as a member of the cadherin family of cell adhesion molecules. Eur J Cell Biol. 1990 Oct;53(1):1–12. [PubMed] [Google Scholar]
  29. Kowalczyk A. P., Palka H. L., Luu H. H., Nilles L. A., Anderson J. E., Wheelock M. J., Green K. J. Posttranslational regulation of plakoglobin expression. Influence of the desmosomal cadherins on plakoglobin metabolic stability. J Biol Chem. 1994 Dec 9;269(49):31214–31223. [PubMed] [Google Scholar]
  30. Leube R. E., Kaiser P., Seiter A., Zimbelmann R., Franke W. W., Rehm H., Knaus P., Prior P., Betz H., Reinke H. Synaptophysin: molecular organization and mRNA expression as determined from cloned cDNA. EMBO J. 1987 Nov;6(11):3261–3268. doi: 10.1002/j.1460-2075.1987.tb02644.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Leube R. E., Leimer U., Grund C., Franke W. W., Harth N., Wiedenmann B. Sorting of synaptophysin into special vesicles in nonneuroendocrine epithelial cells. J Cell Biol. 1994 Dec;127(6 Pt 1):1589–1601. doi: 10.1083/jcb.127.6.1589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Leube R. E., Wiedenmann B., Franke W. W. Topogenesis and sorting of synaptophysin: synthesis of a synaptic vesicle protein from a gene transfected into nonneuroendocrine cells. Cell. 1989 Nov 3;59(3):433–446. doi: 10.1016/0092-8674(89)90028-7. [DOI] [PubMed] [Google Scholar]
  33. Linstedt A. D., Kelly R. B. Synaptophysin is sorted from endocytotic markers in neuroendocrine PC12 cells but not transfected fibroblasts. Neuron. 1991 Aug;7(2):309–317. doi: 10.1016/0896-6273(91)90269-6. [DOI] [PubMed] [Google Scholar]
  34. Mathur M., Goodwin L., Cowin P. Interactions of the cytoplasmic domain of the desmosomal cadherin Dsg1 with plakoglobin. J Biol Chem. 1994 May 13;269(19):14075–14080. [PubMed] [Google Scholar]
  35. McCrea P. D., Turck C. W., Gumbiner B. A homolog of the armadillo protein in Drosophila (plakoglobin) associated with E-cadherin. Science. 1991 Nov 29;254(5036):1359–1361. doi: 10.1126/science.1962194. [DOI] [PubMed] [Google Scholar]
  36. Nagafuchi A., Takeichi M. Cell binding function of E-cadherin is regulated by the cytoplasmic domain. EMBO J. 1988 Dec 1;7(12):3679–3684. doi: 10.1002/j.1460-2075.1988.tb03249.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Näthke I. S., Hinck L., Swedlow J. R., Papkoff J., Nelson W. J. Defining interactions and distributions of cadherin and catenin complexes in polarized epithelial cells. J Cell Biol. 1994 Jun;125(6):1341–1352. doi: 10.1083/jcb.125.6.1341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ozawa M., Baribault H., Kemler R. The cytoplasmic domain of the cell adhesion molecule uvomorulin associates with three independent proteins structurally related in different species. EMBO J. 1989 Jun;8(6):1711–1717. doi: 10.1002/j.1460-2075.1989.tb03563.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Ozawa M., Ringwald M., Kemler R. Uvomorulin-catenin complex formation is regulated by a specific domain in the cytoplasmic region of the cell adhesion molecule. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4246–4250. doi: 10.1073/pnas.87.11.4246. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Peifer M., Berg S., Reynolds A. B. A repeating amino acid motif shared by proteins with diverse cellular roles. Cell. 1994 Mar 11;76(5):789–791. doi: 10.1016/0092-8674(94)90353-0. [DOI] [PubMed] [Google Scholar]
  41. Peifer M. The product of the Drosophila segment polarity gene armadillo is part of a multi-protein complex resembling the vertebrate adherens junction. J Cell Sci. 1993 Aug;105(Pt 4):993–1000. doi: 10.1242/jcs.105.4.993. [DOI] [PubMed] [Google Scholar]
  42. Peifer M., Wieschaus E. The segment polarity gene armadillo encodes a functionally modular protein that is the Drosophila homolog of human plakoglobin. Cell. 1990 Dec 21;63(6):1167–1176. doi: 10.1016/0092-8674(90)90413-9. [DOI] [PubMed] [Google Scholar]
  43. Rose O., Grund C., Reinhardt S., Starzinski-Powitz A., Franke W. W. Contactus adherens, a special type of plaque-bearing adhering junction containing M-cadherin, in the granule cell layer of the cerebellar glomerulus. Proc Natl Acad Sci U S A. 1995 Jun 20;92(13):6022–6026. doi: 10.1073/pnas.92.13.6022. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Rosenthal E. Identification of homologues to beta-catenin/plakoglobin/armadillo in two invertebrates, Urechis caupo and Tripneustes gratilla. Biochim Biophys Acta. 1993 Jun 25;1173(3):337–341. doi: 10.1016/0167-4781(93)90134-y. [DOI] [PubMed] [Google Scholar]
  45. Rubinfeld B., Souza B., Albert I., Munemitsu S., Polakis P. The APC protein and E-cadherin form similar but independent complexes with alpha-catenin, beta-catenin, and plakoglobin. J Biol Chem. 1995 Mar 10;270(10):5549–5555. doi: 10.1074/jbc.270.10.5549. [DOI] [PubMed] [Google Scholar]
  46. Sacco P. A., McGranahan T. M., Wheelock M. J., Johnson K. R. Identification of plakoglobin domains required for association with N-cadherin and alpha-catenin. J Biol Chem. 1995 Aug 25;270(34):20201–20206. doi: 10.1074/jbc.270.34.20201. [DOI] [PubMed] [Google Scholar]
  47. Schmelz M., Duden R., Cowin P., Franke W. W. A constitutive transmembrane glycoprotein of Mr 165,000 (desmoglein) in epidermal and non-epidermal desmosomes. I. Biochemical identification of the polypeptide. Eur J Cell Biol. 1986 Dec;42(2):177–183. [PubMed] [Google Scholar]
  48. Schmidt A., Heid H. W., Schäfer S., Nuber U. A., Zimbelmann R., Franke W. W. Desmosomes and cytoskeletal architecture in epithelial differentiation: cell type-specific plaque components and intermediate filament anchorage. Eur J Cell Biol. 1994 Dec;65(2):229–245. [PubMed] [Google Scholar]
  49. Schwarz M. A., Owaribe K., Kartenbeck J., Franke W. W. Desmosomes and hemidesmosomes: constitutive molecular components. Annu Rev Cell Biol. 1990;6:461–491. doi: 10.1146/annurev.cb.06.110190.002333. [DOI] [PubMed] [Google Scholar]
  50. Schäfer S., Koch P. J., Franke W. W. Identification of the ubiquitous human desmoglein, Dsg2, and the expression catalogue of the desmoglein subfamily of desmosomal cadherins. Exp Cell Res. 1994 Apr;211(2):391–399. doi: 10.1006/excr.1994.1103. [DOI] [PubMed] [Google Scholar]
  51. Schäfer S., Troyanovsky S. M., Heid H. W., Eshkind L. G., Koch P. J., Franke W. W. Cytoskeletal architecture and epithelial differentiation: molecular determinants of cell interaction and cytoskeletal filament anchorage. C R Acad Sci III. 1993 Nov;316(11):1316–1323. [PubMed] [Google Scholar]
  52. Shibamoto S., Hayakawa M., Takeuchi K., Hori T., Miyazawa K., Kitamura N., Johnson K. R., Wheelock M. J., Matsuyoshi N., Takeichi M. Association of p120, a tyrosine kinase substrate, with E-cadherin/catenin complexes. J Cell Biol. 1995 Mar;128(5):949–957. doi: 10.1083/jcb.128.5.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  54. Stappert J., Kemler R. A short core region of E-cadherin is essential for catenin binding and is highly phosphorylated. Cell Adhes Commun. 1994 Aug;2(4):319–327. doi: 10.3109/15419069409014207. [DOI] [PubMed] [Google Scholar]
  55. Su L. K., Vogelstein B., Kinzler K. W. Association of the APC tumor suppressor protein with catenins. Science. 1993 Dec 10;262(5140):1734–1737. doi: 10.1126/science.8259519. [DOI] [PubMed] [Google Scholar]
  56. Takeichi M. Cadherin cell adhesion receptors as a morphogenetic regulator. Science. 1991 Mar 22;251(5000):1451–1455. doi: 10.1126/science.2006419. [DOI] [PubMed] [Google Scholar]
  57. Takeichi M. Cadherins: a molecular family important in selective cell-cell adhesion. Annu Rev Biochem. 1990;59:237–252. doi: 10.1146/annurev.bi.59.070190.001321. [DOI] [PubMed] [Google Scholar]
  58. Troyanovsky S. M., Eshkind L. G., Troyanovsky R. B., Leube R. E., Franke W. W. Contributions of cytoplasmic domains of desmosomal cadherins to desmosome assembly and intermediate filament anchorage. Cell. 1993 Feb 26;72(4):561–574. doi: 10.1016/0092-8674(93)90075-2. [DOI] [PubMed] [Google Scholar]
  59. Troyanovsky S. M., Troyanovsky R. B., Eshkind L. G., Krutovskikh V. A., Leube R. E., Franke W. W. Identification of the plakoglobin-binding domain in desmoglein and its role in plaque assembly and intermediate filament anchorage. J Cell Biol. 1994 Oct;127(1):151–160. doi: 10.1083/jcb.127.1.151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Troyanovsky S. M., Troyanovsky R. B., Eshkind L. G., Leube R. E., Franke W. W. Identification of amino acid sequence motifs in desmocollin, a desmosomal glycoprotein, that are required for plakoglobin binding and plaque formation. Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):10790–10794. doi: 10.1073/pnas.91.23.10790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Wiedenmann B., Franke W. W. Identification and localization of synaptophysin, an integral membrane glycoprotein of Mr 38,000 characteristic of presynaptic vesicles. Cell. 1985 Jul;41(3):1017–1028. doi: 10.1016/s0092-8674(85)80082-9. [DOI] [PubMed] [Google Scholar]

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