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. 1981 Jul 1;90(1):243–248. doi: 10.1083/jcb.90.1.243

Isolation of the intercellular glycoproteins of desmosomes

PMCID: PMC2111822  PMID: 6166625

Abstract

To characterize the desmosome components that mediate intercellular adhesion and cytoskeletal-plasma membrane attachment, we prepared whole desmosomes and isolated desmosomal intercellular regions (desmosomal "cores") from the living cell layers of bovine muzzle epidermis. The tissue was disrupted in a nonionic detergent at low pH, sonicated, and the insoluble residue fractionated by differential centrifugation and metrizamide gradient centrifugation. Transmission electron microscopic analyses reveal that a fraction obtained after differential centrifugation is greatly enriched in whole desmosomes that possess intracellular plaques. Metrizamide gradient centrifugation removes most of the plaque material, leaving the intercellular components and the adjoining plasma membranes. Sodium dodecyl sulfate polyacrylamide gel electrophoresis coupled with methods that reveal carbohydrate- containing moieties on gels demonstrate that certain proteins present in whole desmosomes are glycosylated. These glycoproteins are specifically and greatly enriched in the desmosome cores of which they are the principal protein constituents, and thus may function as the intercellular adhesive of the desmosome.

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Selected References

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  1. Allen T. D., Potten C. S. Desmosomal form, fate, and function in mammalian epidermis. J Ultrastruct Res. 1975 Apr;51(1):94–105. doi: 10.1016/s0022-5320(75)80011-6. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Eckhardt A. E., Hayes C. E., Goldstein I. J. A sensitive fluorescent method for the detection of glycoproteins in polyacrylamide gels. Anal Biochem. 1976 May 21;73(1):192–197. doi: 10.1016/0003-2697(76)90154-8. [DOI] [PubMed] [Google Scholar]
  4. Franke W. W., Weber K., Osborn M., Schmid E., Freudenstein C. Antibody to prekeratin. Decoration of tonofilament like arrays in various cells of epithelial character. Exp Cell Res. 1978 Oct 15;116(2):429–445. doi: 10.1016/0014-4827(78)90466-4. [DOI] [PubMed] [Google Scholar]
  5. Hausman R. E., Moscona A. A. Purification and characterization of the retina-specific cell-aggregating factor. Proc Natl Acad Sci U S A. 1975 Mar;72(3):916–920. doi: 10.1073/pnas.72.3.916. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Henkart P., Humphreys S., Humphreys T. Characterization of sponge aggregation factor. A unique proteoglycan complex. Biochemistry. 1973 Jul 31;12(16):3045–3050. doi: 10.1021/bi00740a016. [DOI] [PubMed] [Google Scholar]
  7. Hopwood D., Logan K. R., Milne G. Mucosubstances in the normal human oesophageal epithelium. A comparison of periodic acid-silver and phophotungstic acid techniques. Histochemistry. 1977 Oct 3;54(1):67–74. doi: 10.1007/BF00493330. [DOI] [PubMed] [Google Scholar]
  8. Hüttermann A., Wendlberger-Schieweg G. Studies on metrizamide-protein interactions. Biochim Biophys Acta. 1976 Nov 26;453(1):176–184. doi: 10.1016/0005-2795(76)90261-0. [DOI] [PubMed] [Google Scholar]
  9. Kelly D. E. Fine structure of desmosomes. , hemidesmosomes, and an adepidermal globular layer in developing newt epidermis. J Cell Biol. 1966 Jan;28(1):51–72. doi: 10.1083/jcb.28.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kelly D. E., Shienvold F. L. The desmosome: fine structural studies with freeze-fracture replication and tannic acid staining of sectioned epidermis. Cell Tissue Res. 1976 Sep 20;172(3):309–323. doi: 10.1007/BF00399514. [DOI] [PubMed] [Google Scholar]
  11. Krawczyk W. S., Wilgram G. F. Hemidesmosome and desmosome morphogenesis during epidermal wound healing. J Ultrastruct Res. 1973 Oct;45(1):93–101. doi: 10.1016/s0022-5320(73)90035-x. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. 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]
  15. McNutt N. S., Weinstein R. S. Membrane ultrastructure at mammalian intercellular junctions. Prog Biophys Mol Biol. 1973;26:45–101. doi: 10.1016/0079-6107(73)90017-5. [DOI] [PubMed] [Google Scholar]
  16. Müller K., Gerisch G. A specific glycoprotein as the target site of adhesion blocking Fab in aggregating Dictyostelium cells. Nature. 1978 Aug 3;274(5670):445–449. doi: 10.1038/274445a0. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Overton J. Formation of junctions and cell sorting in aggregates of chick and mouse cells. Dev Biol. 1977 Jan;55(1):103–116. doi: 10.1016/0012-1606(77)90323-2. [DOI] [PubMed] [Google Scholar]
  19. Rambourg A., Leblond C. P. Electron microscope observations on the carbohydrate-rich cell coat present at the surface of cells in the rat. J Cell Biol. 1967 Jan;32(1):27–53. doi: 10.1083/jcb.32.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rayns D. G., Simpson F. O., Ledingham J. M. Ultrastructure of desmosomes in mammalian intercalated disc; appearances after lanthanum treatment. J Cell Biol. 1969 Jul;42(1):322–326. doi: 10.1083/jcb.42.1.322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rickwood D., Birnie G. D. Metrizamide, a new density-gradient medium. FEBS Lett. 1975 Feb 1;50(2):102–110. doi: 10.1016/0014-5793(75)80467-4. [DOI] [PubMed] [Google Scholar]
  22. Rickwood D., Hell A., Birnie G. D., Gilhuus-Moe C. C. Reversible interaction of metrizamide with protein. Biochim Biophys Acta. 1974 Apr 11;342(2):367–371. doi: 10.1016/0005-2795(74)90092-0. [DOI] [PubMed] [Google Scholar]
  23. Rutishauser U., Thiery J. P., Brackenbury R., Sela B. A., Edelman G. M. Mechanisms of adhesion among cells from neural tissues of the chick embryo. Proc Natl Acad Sci U S A. 1976 Feb;73(2):577–581. doi: 10.1073/pnas.73.2.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Skerrow C. J., Matoltsy A. G. Chemical characterization of isolated epidermal desmosomes. J Cell Biol. 1974 Nov;63(2 Pt 1):524–530. doi: 10.1083/jcb.63.2.524. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Skerrow C. J., Matoltsy A. G. Isolation of epidermal desmosomes. J Cell Biol. 1974 Nov;63(2 Pt 1):515–523. doi: 10.1083/jcb.63.2.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Skerrow C. J. Selective extraction of desmosomal proteins by low ionic strength media. Biochim Biophys Acta. 1979 Jul 25;579(1):241–245. doi: 10.1016/0005-2795(79)90103-x. [DOI] [PubMed] [Google Scholar]
  27. Steinert P. M. The extraction and characterization of bovine epidermal alpha-keratin. Biochem J. 1975 Jul;149(1):39–48. doi: 10.1042/bj1490039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Takeichi M. Functional correlation between cell adhesive properties and some cell surface proteins. J Cell Biol. 1977 Nov;75(2 Pt 1):464–474. doi: 10.1083/jcb.75.2.464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Weinstein R. S., Merk F. B., Alroy J. The structure and function of intercellular junctions in cancer. Adv Cancer Res. 1976;23:23–89. doi: 10.1016/s0065-230x(08)60543-6. [DOI] [PubMed] [Google Scholar]

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