Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1992 Mar 2;116(6):1497–1506. doi: 10.1083/jcb.116.6.1497

Identification of a new hemidesmosomal protein, HD1: a major, high molecular mass component of isolated hemidesmosomes

PMCID: PMC2289367  PMID: 1541639

Abstract

Hemidesmosomes (HDs) mediate cell adhesion to the extracellular matrix and have morphological association with intermediate-sized filaments (IFs) through cytoplasmic plaques. Though several proteins have been located in HDs, most of them have not been well characterized, with the exception of the 230-kD antigen of bullous pemphigoid (BP), an autoimmune skin blistering disease. Only recently we have succeeded in isolating HDs from bovine corneal epithelial cells and in identifying five major components on SDS-PAGE (Owaribe K., Y. Nishizawa, and W. W. Franke. 1991. Exp. Cell Res. 192:622-630). In this study we report on immunological characterization of one of the major components, termed HD1, with an apparent molecular mass of 500 kD. Immunofluorescence microscopy showed colocalization of HD1 with BP antigen at the basement membrane zone of those tissues that have typical HDs, including skin epidermis, corneal and tracheal epithelia, and myoepithelium. In cultured keratinocytes, HD1 demonstrated colocalization with BP antigen in the precise way, while being absent from focal adhesions. Immunoelectron microscopy revealed that an epitope of HD1 was located on the cytoplasmic side of HDs. Taking all these results together, we conclude that HD1 is a new hemidesmosomal component. Interestingly, HD1 also exists in endothelial and glial cells, which lack typical HDs.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

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

  1. Beckerle M. C., Yeh R. K. Talin: role at sites of cell-substratum adhesion. Cell Motil Cytoskeleton. 1990;16(1):7–13. doi: 10.1002/cm.970160103. [DOI] [PubMed] [Google Scholar]
  2. Burridge K., Fath K., Kelly T., Nuckolls G., Turner C. Focal adhesions: transmembrane junctions between the extracellular matrix and the cytoskeleton. Annu Rev Cell Biol. 1988;4:487–525. doi: 10.1146/annurev.cb.04.110188.002415. [DOI] [PubMed] [Google Scholar]
  3. Carter W. G., Kaur P., Gil S. G., Gahr P. J., Wayner E. A. Distinct functions for integrins alpha 3 beta 1 in focal adhesions and alpha 6 beta 4/bullous pemphigoid antigen in a new stable anchoring contact (SAC) of keratinocytes: relation to hemidesmosomes. J Cell Biol. 1990 Dec;111(6 Pt 2):3141–3154. doi: 10.1083/jcb.111.6.3141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Damsky C. H., Knudsen K. A., Bradley D., Buck C. A., Horwitz A. F. Distribution of the cell substratum attachment (CSAT) antigen on myogenic and fibroblastic cells in culture. J Cell Biol. 1985 May;100(5):1528–1539. doi: 10.1083/jcb.100.5.1528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Franke W. W., Schmid E., Freudenstein C., Appelhans B., Osborn M., Weber K., Keenan T. W. Intermediate-sized filaments of the prekeratin type in myoepithelial cells. J Cell Biol. 1980 Mar;84(3):633–654. doi: 10.1083/jcb.84.3.633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gipson I. K., Grill S. M., Spurr S. J., Brennan S. J. Hemidesmosome formation in vitro. J Cell Biol. 1983 Sep;97(3):849–857. doi: 10.1083/jcb.97.3.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Giudice G. J., Squiquera H. L., Elias P. M., Diaz L. A. Identification of two collagen domains within the bullous pemphigoid autoantigen, BP180. J Clin Invest. 1991 Feb;87(2):734–738. doi: 10.1172/JCI115054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Hogervorst F., Kuikman I., von dem Borne A. E., Sonnenberg A. Cloning and sequence analysis of beta-4 cDNA: an integrin subunit that contains a unique 118 kd cytoplasmic domain. EMBO J. 1990 Mar;9(3):765–770. doi: 10.1002/j.1460-2075.1990.tb08171.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hynes R. O. Integrins: a family of cell surface receptors. Cell. 1987 Feb 27;48(4):549–554. doi: 10.1016/0092-8674(87)90233-9. [DOI] [PubMed] [Google Scholar]
  12. Jones J. C., Kurpakus M. A., Cooper H. M., Quaranta V. A function for the integrin alpha 6 beta 4 in the hemidesmosome. Cell Regul. 1991 Jun;2(6):427–438. doi: 10.1091/mbc.2.6.427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jones J. C., Yokoo K. M., Goldman R. D. Is the hemidesmosome a half desmosome? An immunological comparison of mammalian desmosomes and hemidesmosomes. Cell Motil Cytoskeleton. 1986;6(6):560–569. doi: 10.1002/cm.970060604. [DOI] [PubMed] [Google Scholar]
  14. Keene D. R., Sakai L. Y., Lunstrum G. P., Morris N. P., Burgeson R. E. Type VII collagen forms an extended network of anchoring fibrils. J Cell Biol. 1987 Mar;104(3):611–621. doi: 10.1083/jcb.104.3.611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kelly T., Molony L., Burridge K. Purification of two smooth muscle glycoproteins related to integrin. Distribution in cultured chicken embryo fibroblasts. J Biol Chem. 1987 Dec 15;262(35):17189–17199. [PubMed] [Google Scholar]
  16. King S. M., Witman G. B. Structure of the alpha and beta heavy chains of the outer arm dynein from Chlamydomonas flagella. Masses of chains and sites of ultraviolet-induced vanadate-dependent cleavage. J Biol Chem. 1987 Dec 25;262(36):17596–17604. [PubMed] [Google Scholar]
  17. Klatte D. H., Kurpakus M. A., Grelling K. A., Jones J. C. Immunochemical characterization of three components of the hemidesmosome and their expression in cultured epithelial cells. J Cell Biol. 1989 Dec;109(6 Pt 2):3377–3390. doi: 10.1083/jcb.109.6.3377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. Kurpakus M. A., Jones J. C. A novel hemidesmosomal plaque component: tissue distribution and incorporation into assembling hemidesmosomes in an in vitro model. Exp Cell Res. 1991 May;194(1):139–146. doi: 10.1016/0014-4827(91)90143-i. [DOI] [PubMed] [Google Scholar]
  21. Kurpakus M. A., Stock E. L., Jones J. C. Analysis of wound healing in an in vitro model: early appearance of laminin and a 125 x 10(3) Mr polypeptide during adhesion complex formation. J Cell Sci. 1990 Aug;96(Pt 4):651–660. doi: 10.1242/jcs.96.4.651. [DOI] [PubMed] [Google Scholar]
  22. Kyhse-Andersen J. Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J Biochem Biophys Methods. 1984 Dec;10(3-4):203–209. doi: 10.1016/0165-022x(84)90040-x. [DOI] [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. 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]
  25. Mueller S., Klaus-Kovtun V., Stanley J. R. A 230-kD basic protein is the major bullous pemphigoid antigen. J Invest Dermatol. 1989 Jan;92(1):33–38. doi: 10.1111/1523-1747.ep13070476. [DOI] [PubMed] [Google Scholar]
  26. Mutasim D. F., Takahashi Y., Labib R. S., Anhalt G. J., Patel H. P., Diaz L. A. A pool of bullous pemphigoid antigen(s) is intracellular and associated with the basal cell cytoskeleton-hemidesmosome complex. J Invest Dermatol. 1985 Jan;84(1):47–53. doi: 10.1111/1523-1747.ep12274684. [DOI] [PubMed] [Google Scholar]
  27. Otto J. J. Vinculin. Cell Motil Cytoskeleton. 1990;16(1):1–6. doi: 10.1002/cm.970160102. [DOI] [PubMed] [Google Scholar]
  28. Owaribe K., Kartenbeck J., Stumpp S., Magin T. M., Krieg T., Diaz L. A., Franke W. W. The hemidesmosomal plaque. I. Characterization of a major constituent protein as a differentiation marker for certain forms of epithelia. Differentiation. 1990 Dec;45(3):207–220. doi: 10.1111/j.1432-0436.1990.tb00475.x. [DOI] [PubMed] [Google Scholar]
  29. Owaribe K., Nishizawa Y., Franke W. W. Isolation and characterization of hemidesmosomes from bovine corneal epithelial cells. Exp Cell Res. 1991 Feb;192(2):622–630. doi: 10.1016/0014-4827(91)90084-8. [DOI] [PubMed] [Google Scholar]
  30. Regnier M., Vaigot P., Michel S., Prunieras M. Localization of bullous pemphigoid antigen (BPA) in isolated human keratinocytes. J Invest Dermatol. 1985 Sep;85(3):187–190. doi: 10.1111/1523-1747.ep12276656. [DOI] [PubMed] [Google Scholar]
  31. Ruoslahti E., Pierschbacher M. D. New perspectives in cell adhesion: RGD and integrins. Science. 1987 Oct 23;238(4826):491–497. doi: 10.1126/science.2821619. [DOI] [PubMed] [Google Scholar]
  32. Sakai L. Y., Keene D. R., Morris N. P., Burgeson R. E. Type VII collagen is a major structural component of anchoring fibrils. J Cell Biol. 1986 Oct;103(4):1577–1586. doi: 10.1083/jcb.103.4.1577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. 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]
  35. Sonnenberg A., Calafat J., Janssen H., Daams H., van der Raaij-Helmer L. M., Falcioni R., Kennel S. J., Aplin J. D., Baker J., Loizidou M. Integrin alpha 6/beta 4 complex is located in hemidesmosomes, suggesting a major role in epidermal cell-basement membrane adhesion. J Cell Biol. 1991 May;113(4):907–917. doi: 10.1083/jcb.113.4.907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sonnenberg A., Linders C. J., Daams J. H., Kennel S. J. The alpha 6 beta 1 (VLA-6) and alpha 6 beta 4 protein complexes: tissue distribution and biochemical properties. J Cell Sci. 1990 Jun;96(Pt 2):207–217. doi: 10.1242/jcs.96.2.207. [DOI] [PubMed] [Google Scholar]
  37. 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]
  38. Stanley J. R., Hawley-Nelson P., Yuspa S. H., Shevach E. M., Katz S. I. Characterization of bullous pemphigoid antigen: a unique basement membrane protein of stratified squamous epithelia. Cell. 1981 Jun;24(3):897–903. doi: 10.1016/0092-8674(81)90115-x. [DOI] [PubMed] [Google Scholar]
  39. Stanley J. R., Tanaka T., Mueller S., Klaus-Kovtun V., Roop D. Isolation of complementary DNA for bullous pemphigoid antigen by use of patients' autoantibodies. J Clin Invest. 1988 Dec;82(6):1864–1870. doi: 10.1172/JCI113803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Stepp M. A., Spurr-Michaud S., Tisdale A., Elwell J., Gipson I. K. Alpha 6 beta 4 integrin heterodimer is a component of hemidesmosomes. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8970–8974. doi: 10.1073/pnas.87.22.8970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Sugi T., Hashimoto T., Hibi T., Nishikawa T. Production of human monoclonal anti-basement membrane zone (BMZ) antibodies from a patient with bullous pemphigoid (BP) by Epstein-Barr virus transformation. Analyses of the heterogeneity of anti-BMZ antibodies in BP sera using them. J Clin Invest. 1989 Oct;84(4):1050–1055. doi: 10.1172/JCI114266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Suzuki S., Naitoh Y. Amino acid sequence of a novel integrin beta 4 subunit and primary expression of the mRNA in epithelial cells. EMBO J. 1990 Mar;9(3):757–763. doi: 10.1002/j.1460-2075.1990.tb08170.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Takahashi Y., Mutasim D. F., Patel H. P., Anhalt G. J., Labib R. S., Diaz L. A. The use of human pemphigoid autoantibodies to study the fate of epidermal basal cell hemidesmosomes after trypsin dissociation. J Invest Dermatol. 1985 Oct;85(4):309–313. doi: 10.1111/1523-1747.ep12276893. [DOI] [PubMed] [Google Scholar]
  44. Tamura R. N., Rozzo C., Starr L., Chambers J., Reichardt L. F., Cooper H. M., Quaranta V. Epithelial integrin alpha 6 beta 4: complete primary structure of alpha 6 and variant forms of beta 4. J Cell Biol. 1990 Oct;111(4):1593–1604. doi: 10.1083/jcb.111.4.1593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Thacher S. M., Malone K. L., Dave K., Zhao S. M. Localization of the 230-kilodalton bullous pemphigoid antigen in cultured keratinocytes: formation of a prehemidesmosome. Exp Cell Res. 1991 Jun;194(2):238–247. doi: 10.1016/0014-4827(91)90360-7. [DOI] [PubMed] [Google Scholar]
  46. 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]
  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]
  48. Tsukita S., Tsukita S., Nagafuchi A. The undercoat of adherens junctions: a key specialized structure in organogenesis and carcinogenesis. Cell Struct Funct. 1990 Feb;15(1):7–12. doi: 10.1247/csf.15.7. [DOI] [PubMed] [Google Scholar]
  49. Verrando P., Hsi B. L., Yeh C. J., Pisani A., Serieys N., Ortonne J. P. Monoclonal antibody GB3, a new probe for the study of human basement membranes and hemidesmosomes. Exp Cell Res. 1987 May;170(1):116–128. doi: 10.1016/0014-4827(87)90121-2. [DOI] [PubMed] [Google Scholar]
  50. Westgate G. E., Weaver A. C., Couchman J. R. Bullous pemphigoid antigen localization suggests an intracellular association with hemidesmosomes. J Invest Dermatol. 1985 Mar;84(3):218–224. doi: 10.1111/1523-1747.ep12265229. [DOI] [PubMed] [Google Scholar]

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

RESOURCES