Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1985 Dec 1;101(6):2070–2080. doi: 10.1083/jcb.101.6.2070

Desmocalmin: a calmodulin-binding high molecular weight protein isolated from desmosomes

PMCID: PMC2114022  PMID: 2415534

Abstract

A unique high molecular weight protein (240,000 mol wt) has been purified from isolated desmosomes of bovine muzzle epidermis, using low- salt extraction at pH 9.5-10.5 and gel-filtration followed by calmodulin-affinity column chromatography. This protein was shown to bind to calmodulin in a Ca2+-dependent manner, so we called it desmocalmin here. Desmocalmin also bound to the reconstituted keratin filaments in vitro in the presence of Mg2+, but not to actin filaments. By use of the antibody raised against the purified desmocalmin, desmocalmin was shown by both immunoelectron and immunofluorescence microscopy to be localized at the desmosomal plaque just beneath the plasma membrane. Judging from its isoelectric point and antigenicity, desmocalmin was clearly distinct from desmoplakins I and II, which were identified in the desmosomal plaque by Mueller and Franke (1983, J. Mol. Biol., 163:647-671). In the low-angle, rotary-shadowing electron microscope, the desmocalmin molecules looked like flexible rods approximately 100-nm long consisting of two polypeptide chains lying side by side. The similar rodlike structures were clearly identified in the freeze-etch replica images of desmosomes. Taken together, these findings indicate that desmocalmin could function as a key protein responsible for the formation of desmosomes in a calmodulin-dependent manner (Trinkaus-Randall, V., and I.K. Gipson, 1984, J. Cell Biol., 98:1565-1571).

Full Text

The Full Text of this article is available as a PDF (2.1 MB).

Selected References

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

  1. 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]
  2. 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]
  3. Dembitzer H. M., Herz F., Schermer A., Wolley R. C., Koss L. G. Desmosome development in an in vitro model. J Cell Biol. 1980 Jun;85(3):695–702. doi: 10.1083/jcb.85.3.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Franke W. W., Heid H. W., Grund C., Winter S., Freudenstein C., Schmid E., Jarasch E. D., Keenan T. W. Antibodies to the major insoluble milk fat globule membrane-associated protein: specific location in apical regions of lactating epithelial cells. J Cell Biol. 1981 Jun;89(3):485–494. doi: 10.1083/jcb.89.3.485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Geiger B., Avnur Z., Rinnerthaler G., Hinssen H., Small V. J. Microfilament-organizing centers in areas of cell contact: cytoskeletal interactions during cell attachment and locomotion. J Cell Biol. 1984 Jul;99(1 Pt 2):83s–91s. doi: 10.1083/jcb.99.1.83s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Glenney J. R., Jr, Glenney P., Osborn M., Weber K. An F-actin- and calmodulin-binding protein from isolated intestinal brush borders has a morphology related to spectrin. Cell. 1982 Apr;28(4):843–854. doi: 10.1016/0092-8674(82)90063-0. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. 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]
  12. 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]
  13. Jarasch E. D., Grund C., Bruder G., Heid H. W., Keenan T. W., Franke W. W. Localization of xanthine oxidase in mammary-gland epithelium and capillary endothelium. Cell. 1981 Jul;25(1):67–82. doi: 10.1016/0092-8674(81)90232-4. [DOI] [PubMed] [Google Scholar]
  14. Jones J. C., Goldman A. E., Steinert P. M., Yuspa S., Goldman R. D. Dynamic aspects of the supramolecular organization of intermediate filament networks in cultured epidermal cells. Cell Motil. 1982;2(3):197–213. doi: 10.1002/cm.970020302. [DOI] [PubMed] [Google Scholar]
  15. Kakiuchi S., Sobue K., Fujita M. Purification of a 240 000 Mr calmodulin-binding protein from a microsomal fraction of brain. FEBS Lett. 1981 Sep 14;132(1):144–148. doi: 10.1016/0014-5793(81)80449-8. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Lazarides E., Nelson W. J. Expression of spectrin in nonerythroid cells. Cell. 1982 Dec;31(3 Pt 2):505–508. doi: 10.1016/0092-8674(82)90306-3. [DOI] [PubMed] [Google Scholar]
  18. Levine J., Willard M. Fodrin: axonally transported polypeptides associated with the internal periphery of many cells. J Cell Biol. 1981 Sep;90(3):631–642. doi: 10.1083/jcb.90.3.631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mangeat P., Burridge K. Actin-membrane interaction in fibroblasts: what proteins are involved in this association? J Cell Biol. 1984 Jul;99(1 Pt 2):95s–103s. doi: 10.1083/jcb.99.1.95s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Oakley B. R., Kirsch D. R., Morris N. R. A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels. Anal Biochem. 1980 Jul 1;105(2):361–363. doi: 10.1016/0003-2697(80)90470-4. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Repasky E. A., Granger B. L., Lazarides E. Widespread occurrence of avian spectrin in nonerythroid cells. Cell. 1982 Jul;29(3):821–833. doi: 10.1016/0092-8674(82)90444-5. [DOI] [PubMed] [Google Scholar]
  24. Shimo-Oka T., Watanabe Y. Stimulation of actomyosin Mg2+-ATPase activity by a brain microtubule-associated protein fraction. High-molecular-weight actin-binding protein is the stimulating factor. J Biochem. 1981 Nov;90(5):1297–1307. doi: 10.1093/oxfordjournals.jbchem.a133595. [DOI] [PubMed] [Google Scholar]
  25. Singer S. J. The molecular organization of membranes. Annu Rev Biochem. 1974;43(0):805–833. doi: 10.1146/annurev.bi.43.070174.004105. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. 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]
  28. Spudich J. A., Watt S. The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. J Biol Chem. 1971 Aug 10;246(15):4866–4871. [PubMed] [Google Scholar]
  29. Staehelin L. A. Structure and function of intercellular junctions. Int Rev Cytol. 1974;39:191–283. doi: 10.1016/s0074-7696(08)60940-7. [DOI] [PubMed] [Google Scholar]
  30. Steinert P. M., Idler W. W., Zimmerman S. B. Self-assembly of bovine epidermal keratin filaments in vitro. J Mol Biol. 1976 Dec 15;108(3):547–567. doi: 10.1016/s0022-2836(76)80136-2. [DOI] [PubMed] [Google Scholar]
  31. Trinkaus-Randall V., Gipson I. K. Role of calcium and calmodulin in hemidesmosome formation in vitro. J Cell Biol. 1984 Apr;98(4):1565–1571. doi: 10.1083/jcb.98.4.1565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tsukita S., Tsukita S., Ishikawa H. Association of actin and 10 nm filaments with the dense body in smooth muscle cells of the chicken gizzard. Cell Tissue Res. 1983;229(2):233–242. doi: 10.1007/BF00214972. [DOI] [PubMed] [Google Scholar]
  33. Tsukita S., Tsukita S., Ishikawa H. Cytoskeletal network underlying the human erythrocyte membrane. Thin-section electron microscopy. J Cell Biol. 1980 Jun;85(3):567–576. doi: 10.1083/jcb.85.3.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tsukita S., Tsukita S., Ishikawa H., Kurokawa M., Morimoto K., Sobue K., Kakiuchi S. Binding sites of calmodulin and actin on the brain spectrin, calspectin. J Cell Biol. 1983 Aug;97(2):574–578. doi: 10.1083/jcb.97.2.574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tsukita S., Tsukita S., Ishikawa H., Sato S., Nakao M. Electron microscopic study of reassociation of spectrin and actin with the human erythrocyte membrane. J Cell Biol. 1981 Jul;90(1):70–77. doi: 10.1083/jcb.90.1.70. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Tsukita S., Tsukita S., Usukura J., Ishikawa H. ATP-dependent structural changes of the outer dynein arm in Tetrahymena cilia: a freeze-etch replica study. J Cell Biol. 1983 May;96(5):1480–1485. doi: 10.1083/jcb.96.5.1480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Tyler J. M., Branton D. Rotary shadowing of extended molecules dried from glycerol. J Ultrastruct Res. 1980 May;71(2):95–102. doi: 10.1016/s0022-5320(80)90098-2. [DOI] [PubMed] [Google Scholar]
  38. 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]
  39. 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]
  40. Wiche G., Krepler R., Artlieb U., Pytela R., Denk H. Occurrence and immunolocalization of plectin in tissues. J Cell Biol. 1983 Sep;97(3):887–901. doi: 10.1083/jcb.97.3.887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wiseman L. L., Strickler J. Desmosome frequency: Experimental alteration may correlate with differential cell adhesion. J Cell Sci. 1981 Jun;49:217–223. doi: 10.1242/jcs.49.1.217. [DOI] [PubMed] [Google Scholar]

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

RESOURCES