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. 1992 Sep 2;118(6):1477–1488. doi: 10.1083/jcb.118.6.1477

Identification of an epithelial protein related to the desmosome and intermediate filament network

PMCID: PMC2289612  PMID: 1522118

Abstract

Using a mAb, referred to as 08L, we have identified a protein, of M(r) approximately 140,000, associated with desmosomes of epithelial cells. The 08L antibody stained the intracellular side of lateral cell margins of monolayer epithelial cells but did not stain cell margins free of cell contact. Immunoelectron microscopy revealed that the 08L antigen was localized to the cytosolic surface of the desmosomal plaque near points of intermediate filament convergence with apparently little staining of the desmosomal plaque proper. Western blots revealed the 08L antigen to be a protein, of M(r) approximately 140,000, found in the Triton-X 100 insoluble pellet. High salt-containing buffers extracted the 08L antigen from the insoluble material. Examination of the assembly of 08L to the desmosome complex, in cells grown in low confluent culture or in calcium-switch assays, by double immunofluorescence with 08L and anti-desmoplakin antibody, revealed that 08L was recruited to morphologically identifiable desmosomes. 08L antigen may exist in a cytosolic pool prior to assembly to the cell surface. The solubility of 08L in low calcium and normal calcium conditions, however, was similar. 08L association to the desmosome was correlated with increased organization of the intermediate filament network. We suggest that the 08L antigen may be involved in the organization and stabilization of the desmosome-IF complexes of epithelia.

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Selected References

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  1. Bologna M., Allen R., Dulbecco R. Organization of cytokeratin bundles by desmosomes in rat mammary cells. J Cell Biol. 1986 Feb;102(2):560–567. doi: 10.1083/jcb.102.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cartaud A., Ludosky M. A., Courvalin J. C., Cartaud J. A protein antigenically related to nuclear lamin B mediates the association of intermediate filaments with desmosomes. J Cell Biol. 1990 Aug;111(2):581–588. doi: 10.1083/jcb.111.2.581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. Cowin P., Mattey D., Garrod D. Identification of desmosomal surface components (desmocollins) and inhibition of desmosome formation by specific Fab'. J Cell Sci. 1984 Aug;70:41–60. doi: 10.1242/jcs.70.1.41. [DOI] [PubMed] [Google Scholar]
  6. Dodson J. W., Hay E. D. Secretion of collagenous stroma by isolated epithelium grown in vitro. Exp Cell Res. 1971 Mar;65(1):215–220. doi: 10.1016/s0014-4827(71)80069-1. [DOI] [PubMed] [Google Scholar]
  7. Drochmans P., Freudenstein C., Wanson J. C., Laurent L., Keenan T. W., Stadler J., Leloup R., Franke W. W. Structure and biochemical composition of desmosomes and tonofilaments isolated from calf muzzle epidermis. J Cell Biol. 1978 Nov;79(2 Pt 1):427–443. doi: 10.1083/jcb.79.2.427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Duden R., Franke W. W. Organization of desmosomal plaque proteins in cells growing at low calcium concentrations. J Cell Biol. 1988 Sep;107(3):1049–1063. doi: 10.1083/jcb.107.3.1049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fey E. G., Wan K. M., Penman S. Epithelial cytoskeletal framework and nuclear matrix-intermediate filament scaffold: three-dimensional organization and protein composition. J Cell Biol. 1984 Jun;98(6):1973–1984. doi: 10.1083/jcb.98.6.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Geiger B., Ginsberg D. The cytoplasmic domain of adherens-type junctions. Cell Motil Cytoskeleton. 1991;20(1):1–6. doi: 10.1002/cm.970200102. [DOI] [PubMed] [Google Scholar]
  12. Gorbsky G., Cohen S. M., Shida H., Giudice G. J., Steinberg M. S. Isolation of the non-glycosylated proteins of desmosomes and immunolocalization of a third plaque protein: desmoplakin III. Proc Natl Acad Sci U S A. 1985 Feb;82(3):810–814. doi: 10.1073/pnas.82.3.810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Green K. J., Parry D. A., Steinert P. M., Virata M. L., Wagner R. M., Angst B. D., Nilles L. A. Structure of the human desmoplakins. Implications for function in the desmosomal plaque. J Biol Chem. 1990 Feb 15;265(5):2603–2612. [PubMed] [Google Scholar]
  14. Hennings H., Michael D., Cheng C., Steinert P., Holbrook K., Yuspa S. H. Calcium regulation of growth and differentiation of mouse epidermal cells in culture. Cell. 1980 Jan;19(1):245–254. doi: 10.1016/0092-8674(80)90406-7. [DOI] [PubMed] [Google Scholar]
  15. Hieda Y., Tsukita S., Tsukita S. A new high molecular mass protein showing unique localization in desmosomal plaque. J Cell Biol. 1989 Oct;109(4 Pt 1):1511–1518. doi: 10.1083/jcb.109.4.1511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Holton J. L., Kenny T. P., Legan P. K., Collins J. E., Keen J. N., Sharma R., Garrod D. R. Desmosomal glycoproteins 2 and 3 (desmocollins) show N-terminal similarity to calcium-dependent cell-cell adhesion molecules. J Cell Sci. 1990 Oct;97(Pt 2):239–246. doi: 10.1242/jcs.97.2.239. [DOI] [PubMed] [Google Scholar]
  17. Inagaki M., Nishi Y., Nishizawa K., Matsuyama M., Sato C. Site-specific phosphorylation induces disassembly of vimentin filaments in vitro. Nature. 1987 Aug 13;328(6131):649–652. doi: 10.1038/328649a0. [DOI] [PubMed] [Google Scholar]
  18. Jones J. C. Characterization of a 125K glycoprotein associated with bovine epithelial desmosomes. J Cell Sci. 1988 Feb;89(Pt 2):207–216. doi: 10.1242/jcs.89.2.207. [DOI] [PubMed] [Google Scholar]
  19. Jones J. C., Goldman R. D. Intermediate filaments and the initiation of desmosome assembly. J Cell Biol. 1985 Aug;101(2):506–517. doi: 10.1083/jcb.101.2.506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jones J. C., Green K. J. Intermediate filament-plasma membrane interactions. Curr Opin Cell Biol. 1991 Feb;3(1):127–132. doi: 10.1016/0955-0674(91)90175-x. [DOI] [PubMed] [Google Scholar]
  21. Jones J. C., Yokoo K. M., Goldman R. D. A cell surface desmosome-associated component: identification of tissue-specific cell adhesion molecule. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7282–7286. doi: 10.1073/pnas.83.19.7282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kapprell H. P., Owaribe K., Franke W. W. Identification of a basic protein of Mr 75,000 as an accessory desmosomal plaque protein in stratified and complex epithelia. J Cell Biol. 1988 May;106(5):1679–1691. doi: 10.1083/jcb.106.5.1679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Lentz T. L., Trinkaus J. P. Differentiation of the junctional complex of surface cells in the developing Fundulus blastoderm. J Cell Biol. 1971 Mar;48(3):455–472. doi: 10.1083/jcb.48.3.455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Magee A. I., Buxton R. S. Transmembrane molecular assemblies regulated by the greater cadherin family. Curr Opin Cell Biol. 1991 Oct;3(5):854–861. doi: 10.1016/0955-0674(91)90060-c. [DOI] [PubMed] [Google Scholar]
  28. Mattey D. L., Garrod D. R. Calcium-induced desmosome formation in cultured kidney epithelial cells. J Cell Sci. 1986 Sep;85:95–111. doi: 10.1242/jcs.85.1.95. [DOI] [PubMed] [Google Scholar]
  29. Mattey D. L., Garrod D. R. Calcium-induced desmosome formation in cultured kidney epithelial cells. J Cell Sci. 1986 Sep;85:95–111. doi: 10.1242/jcs.85.1.95. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Mechanic S., Raynor K., Hill J. E., Cowin P. Desmocollins form a distinct subset of the cadherin family of cell adhesion molecules. Proc Natl Acad Sci U S A. 1991 May 15;88(10):4476–4480. doi: 10.1073/pnas.88.10.4476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Mueller H., Franke W. W. Biochemical and immunological characterization of desmoplakins I and II, the major polypeptides of the desmosomal plaque. J Mol Biol. 1983 Feb 5;163(4):647–671. doi: 10.1016/0022-2836(83)90116-x. [DOI] [PubMed] [Google Scholar]
  33. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  34. O'Keefe E. J., Erickson H. P., Bennett V. Desmoplakin I and desmoplakin II. Purification and characterization. J Biol Chem. 1989 May 15;264(14):8310–8318. [PubMed] [Google Scholar]
  35. OVERTON J. Desmosome development in normal and reassociating cells in the early chick blastoderm. Dev Biol. 1962 Jun;4:532–548. doi: 10.1016/0012-1606(62)90056-8. [DOI] [PubMed] [Google Scholar]
  36. Pasdar M., Krzeminski K. A., Nelson W. J. Regulation of desmosome assembly in MDCK epithelial cells: coordination of membrane core and cytoplasmic plaque domain assembly at the plasma membrane. J Cell Biol. 1991 May;113(3):645–655. doi: 10.1083/jcb.113.3.645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Pasdar M., Nelson W. J. Kinetics of desmosome assembly in Madin-Darby canine kidney epithelial cells: temporal and spatial regulation of desmoplakin organization and stabilization upon cell-cell contact. I. Biochemical analysis. J Cell Biol. 1988 Mar;106(3):677–685. doi: 10.1083/jcb.106.3.677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Pasdar M., Nelson W. J. Kinetics of desmosome assembly in Madin-Darby canine kidney epithelial cells: temporal and spatial regulation of desmoplakin organization and stabilization upon cell-cell contact. II. Morphological analysis. J Cell Biol. 1988 Mar;106(3):687–695. doi: 10.1083/jcb.106.3.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Pasdar M., Nelson W. J. Regulation of desmosome assembly in epithelial cells: kinetics of synthesis, transport, and stabilization of desmoglein I, a major protein of the membrane core domain. J Cell Biol. 1989 Jul;109(1):163–177. doi: 10.1083/jcb.109.1.163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. 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]
  41. Schmelz M., Duden R., Cowin P., Franke W. W. A constitutive transmembrane glycoprotein of Mr 165,000 (desmoglein) in epidermal and non-epidermal desmosomes. II. Immunolocalization and microinjection studies. Eur J Cell Biol. 1986 Dec;42(2):184–199. [PubMed] [Google Scholar]
  42. 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]
  43. Stappenbeck T. S., Green K. J. The desmoplakin carboxyl terminus coaligns with and specifically disrupts intermediate filament networks when expressed in cultured cells. J Cell Biol. 1992 Mar;116(5):1197–1209. doi: 10.1083/jcb.116.5.1197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Steinert P. M., Roop D. R. Molecular and cellular biology of intermediate filaments. Annu Rev Biochem. 1988;57:593–625. doi: 10.1146/annurev.bi.57.070188.003113. [DOI] [PubMed] [Google Scholar]
  45. Suhrbier A., Garrod D. An investigation of the molecular components of desmosomes in epithelial cells of five vertebrates. J Cell Sci. 1986 Mar;81:223–242. doi: 10.1242/jcs.81.1.223. [DOI] [PubMed] [Google Scholar]
  46. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Tsukita S., Tsukita S. Desmocalmin: a calmodulin-binding high molecular weight protein isolated from desmosomes. J Cell Biol. 1985 Dec;101(6):2070–2080. doi: 10.1083/jcb.101.6.2070. [DOI] [PMC free article] [PubMed] [Google Scholar]

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