Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1987 Nov 1;105(5):2327–2334. doi: 10.1083/jcb.105.5.2327

Structure and assembly of desmosome junctions: biosynthesis and turnover of the major desmosome components of Madin-Darby canine kidney cells in low calcium medium

PMCID: PMC2114848  PMID: 3680384

Abstract

Neither stratifying (primary keratinocytes) nor simple (Madin-Darby canine kidney [MDCK] and Madin-Darby bovine kidney [MDBK]) epithelial cell types from desmosomes in low calcium medium (LCM; less than 0.1 mM), but they can be induced to do so by raising the calcium level to physiological concentrations (standard calcium medium [SCM], 2 mM). We have used polyclonal antisera to the major bovine epidermal desmosome components (greater than 100 kD) in a sensitive assay involving immunoprecipitation of the components from metabolically labeled MDCK cell monolayers to investigate the mechanism of calcium-induced desmosome formation. MDCK cells, whether cultured in LCM or SCM, were found to synthesize the desmosome protein, DPI and desmosome glycoproteins DGI and DGII/III with identical electrophoretic mobility, and also, where relevant, with similar carbohydrate addition/processing and proteolytic processing. The timings of these events and of transport of DGI to the cell surface were similar in low and high calcium. Although the rates of synthesis of the various desmosome components were also similar under both conditions, the glycoprotein turnover rates increased dramatically in cells cultured in LCM. The half-lives decreased by a factor of about 7 for DGI and 12 for DGII/III and, consistent with this, MDCK cells labeled for 48 h in SCM had three and six times the amount of DGI and DGII/III, respectively, as cells labeled for 48 h in LCM. The rate of turnover and the levels of DPI were changed in the same direction, but to much lesser extents. Possible mechanisms for the Ca2+-dependent control of desmosome formation are discussed in the light of this new evidence.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

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

  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. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  3. Brennan J. K., Mansky J., Roberts G., Lichtman M. A. Improved methods for reducing calcium and magnesium concentrations in tissue culture medium: application to studies of lymphoblast proliferation in vitro. In Vitro. 1975 Nov-Dec;11(6):354–360. doi: 10.1007/BF02616371. [DOI] [PubMed] [Google Scholar]
  4. Cohen S. M., Gorbsky G., Steinberg M. S. Immunochemical characterization of related families of glycoproteins in desmosomes. J Biol Chem. 1983 Feb 25;258(4):2621–2627. [PubMed] [Google Scholar]
  5. Cowin P., Garrod D. R. Antibodies to epithelial desmosomes show wide tissue and species cross-reactivity. Nature. 1983 Mar 10;302(5904):148–150. doi: 10.1038/302148a0. [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. Cowin P., Mattey D., Garrod D. Distribution of desmosomal components in the tissues of vertebrates, studied by fluorescent antibody staining. J Cell Sci. 1984 Mar;66:119–132. doi: 10.1242/jcs.66.1.119. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Dulbecco R., Allen W. R., Bowman M. Lumen formation and redistribution of inframembranous proteins during differentiation of ducts in the rat mammary gland. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5763–5766. doi: 10.1073/pnas.81.18.5763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Franke W. W., Cowin P., Schmelz M., Kapprell H. P. The desmosomal plaque and the cytoskeleton. Ciba Found Symp. 1987;125:26–48. doi: 10.1002/9780470513408.ch3. [DOI] [PubMed] [Google Scholar]
  11. Franke W. W., Moll R., Mueller H., Schmid E., Kuhn C., Krepler R., Artlieb U., Denk H. Immunocytochemical identification of epithelium-derived human tumors with antibodies to desmosomal plaque proteins. Proc Natl Acad Sci U S A. 1983 Jan;80(2):543–547. doi: 10.1073/pnas.80.2.543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Franke W. W., Moll R., Schiller D. L., Schmid E., Kartenbeck J., Mueller H. Desmoplakins of epithelial and myocardial desmosomes are immunologically and biochemically related. Differentiation. 1982;23(2):115–127. doi: 10.1111/j.1432-0436.1982.tb01274.x. [DOI] [PubMed] [Google Scholar]
  13. Franke W. W., Mueller H., Mittnacht S., Kapprell H. P., Jorcano J. L. Significance of two desmosome plaque-associated polypeptides of molecular weights 75 000 and 83 000. EMBO J. 1983;2(12):2211–2215. doi: 10.1002/j.1460-2075.1983.tb01725.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Franke W. W., Schmid E., Grund C., Müller H., Engelbrecht I., Moll R., Stadler J., Jarasch E. D. Antibodies to high molecular weight polypeptides of desmosomes: specific localization of a class of junctional proteins in cells and tissue. Differentiation. 1981;20(3):217–241. doi: 10.1111/j.1432-0436.1981.tb01178.x. [DOI] [PubMed] [Google Scholar]
  15. Giudice G. J., Cohen S. M., Patel N. H., Steinberg M. S. Immunological comparison of desmosomal components from several bovine tissues. J Cell Biochem. 1984;26(1):35–45. doi: 10.1002/jcb.240260104. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Gorbsky G., Steinberg M. S. Isolation of the intercellular glycoproteins of desmosomes. J Cell Biol. 1981 Jul;90(1):243–248. doi: 10.1083/jcb.90.1.243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hennings H., Holbrook K. A. Calcium regulation of cell-cell contact and differentiation of epidermal cells in culture. An ultrastructural study. Exp Cell Res. 1983 Jan;143(1):127–142. doi: 10.1016/0014-4827(83)90115-5. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. Kartenbeck J., Schmid E., Franke W. W., Geiger B. Different modes of internalization of proteins associated with adhaerens junctions and desmosomes: experimental separation of lateral contacts induces endocytosis of desmosomal plaque material. EMBO J. 1982;1(6):725–732. doi: 10.1002/j.1460-2075.1982.tb01237.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  23. 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]
  24. Lazarides E., Moon R. T. Assembly and topogenesis of the spectrin-based membrane skeleton in erythroid development. Cell. 1984 Jun;37(2):354–356. doi: 10.1016/0092-8674(84)90364-7. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Mattey D. L., Garrod D. R. Splitting and internalization of the desmosomes of cultured kidney epithelial cells by reduction in calcium concentration. J Cell Sci. 1986 Sep;85:113–124. doi: 10.1242/jcs.85.1.113. [DOI] [PubMed] [Google Scholar]
  27. Meldolesi J., Castiglioni G., Parma R., Nassivera N., De Camilli P. Ca++-dependent disassembly and reassembly of occluding junctions in guinea pig pancreatic acinar cells. Effect of drugs. J Cell Biol. 1978 Oct;79(1):156–172. doi: 10.1083/jcb.79.1.156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Miller K., Mattey D., Measures H., Hopkins C., Garrod D. Localisation of the protein and glycoprotein components of bovine nasal epithelial desmosomes by immunoelectron microscopy. EMBO J. 1987 Apr;6(4):885–889. doi: 10.1002/j.1460-2075.1987.tb04834.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Penn E. J., Hobson C., Rees D. A., Magee A. I. Structure and assembly of desmosome junctions: biosynthesis, processing, and transport of the major protein and glycoprotein components in cultured epithelial cells. J Cell Biol. 1987 Jul;105(1):57–68. doi: 10.1083/jcb.105.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pitelka D. R., Taggart B. N., Hamamoto S. T. Effects of extracellular calcium depletion on membrane topography and occluding junctions of mammary epithelial cells in culture. J Cell Biol. 1983 Mar;96(3):613–624. doi: 10.1083/jcb.96.3.613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Richardson J. C., Scalera V., Simmons N. L. Identification of two strains of MDCK cells which resemble separate nephron tubule segments. Biochim Biophys Acta. 1981 Feb 18;673(1):26–36. [PubMed] [Google Scholar]
  32. 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]
  33. Steinberg M. S., Shida H., Giudice G. J., Shida M., Patel N. H., Blaschuk O. W. On the molecular organization, diversity and functions of desmosomal proteins. Ciba Found Symp. 1987;125:3–25. doi: 10.1002/9780470513408.ch2. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. 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]
  36. Volberg T., Geiger B., Kartenbeck J., Franke W. W. Changes in membrane-microfilament interaction in intercellular adherens junctions upon removal of extracellular Ca2+ ions. J Cell Biol. 1986 May;102(5):1832–1842. doi: 10.1083/jcb.102.5.1832. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Watt F. M., Mattey D. L., Garrod D. R. Calcium-induced reorganization of desmosomal components in cultured human keratinocytes. J Cell Biol. 1984 Dec;99(6):2211–2215. doi: 10.1083/jcb.99.6.2211. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES