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. 1982 Mar 1;92(3):665–673. doi: 10.1083/jcb.92.3.665

Monoclonal antibodies provide specific intramolecular markers for the study of epithelial tonofilament organization

PMCID: PMC2112033  PMID: 6177700

Abstract

The tonofilament-associated protein antigens recognized in epithelial cells by a group of six monoclonal antibodies have been studied by immunofluorescence and gel immunoautoradiography. The monoclonal antibodies were generated against detergent insoluble cytoskeleton extracts from a cultured simple epithelium derived cell line, Ptk1 cells. They show various tissue specificities, and while they all recognize components at the low end of the molecular weight range for intermediate filament proteins, they confirm that single antibody species can react with multiple polypeptides of different molecular weights in the tonofilament complex. The monoclonal antibodies described here demonstrate the presence of a simple epithelium antigenic determinant associated with intermediate filaments that is not detectable in the specialized cells of squamous and keratinizing epithelia but can reappear in such cells after transformation.

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Selected References

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  1. Brûlet P., Babinet C., Kemler R., Jacob F. Monoclonal antibodies against trophectoderm-specific markers during mouse blastocyst formation. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4113–4117. doi: 10.1073/pnas.77.7.4113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Burridge K. Direct identification of specific glycoproteins and antigens in sodium dodecyl sulfate gels. Methods Enzymol. 1978;50:54–64. doi: 10.1016/0076-6879(78)50007-4. [DOI] [PubMed] [Google Scholar]
  3. Franke W. W., Grund C., Osborn M., Weber K. The intermediate-sized filaments in rat kangaroo PtK2 cells. I. Morphology in situ. Cytobiologie. 1978 Aug;17(2):365–391. [PubMed] [Google Scholar]
  4. Franke W. W., Schmid E., Osborn M., Weber K. The intermediate-sized filaments in rat kangaroo PtK2 cells. II. Structure and composition of isolated filaments. Cytobiologie. 1978 Aug;17(2):392–411. [PubMed] [Google Scholar]
  5. Franke W. W., Schmid E., Weber K., Osborn M. HeLa cells contain intermediate-sized filaments of the prekeratin type. Exp Cell Res. 1979 Jan;118(1):95–109. doi: 10.1016/0014-4827(79)90587-1. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Fuchs E., Green H. Changes in keratin gene expression during terminal differentiation of the keratinocyte. Cell. 1980 Apr;19(4):1033–1042. doi: 10.1016/0092-8674(80)90094-x. [DOI] [PubMed] [Google Scholar]
  8. Fuchs E., Green H. Multiple keratins of cultured human epidermal cells are translated from different mRNA molecules. Cell. 1979 Jul;17(3):573–582. doi: 10.1016/0092-8674(79)90265-4. [DOI] [PubMed] [Google Scholar]
  9. Fuchs E., Green H. The expression of keratin genes in epidermis and cultured epidermal cells. Cell. 1978 Nov;15(3):887–897. doi: 10.1016/0092-8674(78)90273-8. [DOI] [PubMed] [Google Scholar]
  10. Gordon W. E., 3rd, Bushnell A., Burridge K. Characterization of the intermediate (10 nm) filaments of cultured cells using an autoimmune rabbit antiserum. Cell. 1978 Feb;13(2):249–261. doi: 10.1016/0092-8674(78)90194-0. [DOI] [PubMed] [Google Scholar]
  11. Kemp D. J. Unique and repetitive sequences in multiple genes for feather keratin. Nature. 1975 Apr 17;254(5501):573–577. doi: 10.1038/254573a0. [DOI] [PubMed] [Google Scholar]
  12. Kennett R. H., Denis K. A., Tung A. S., Klinman N. R. Hybrid plasmacytoma production: fusions with adult spleen cells, monoclonal spleen fragments, neonatal spleen cells and human spleen cells. Curr Top Microbiol Immunol. 1978;81:77–91. doi: 10.1007/978-3-642-67448-8_13. [DOI] [PubMed] [Google Scholar]
  13. Kurki P., Linder E., Virtanen I., Stenman S. Human smooth muscle autoantibodies reacting with intermediate (100 A) filaments. Nature. 1977 Jul 21;268(5617):240–241. doi: 10.1038/268240a0. [DOI] [PubMed] [Google Scholar]
  14. Köhler G., Milstein C. Derivation of specific antibody-producing tissue culture and tumor lines by cell fusion. Eur J Immunol. 1976 Jul;6(7):511–519. doi: 10.1002/eji.1830060713. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Lawton A. R., 3rd, Asofsky R., Hylton M. B., Cooper M. D. Suppression of immunoglobulin class synthesis in mice. I. Effects of treatment with antibody to -chain. J Exp Med. 1972 Feb 1;135(2):277–297. doi: 10.1084/jem.135.2.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lazarides E. Intermediate filaments as mechanical integrators of cellular space. Nature. 1980 Jan 17;283(5744):249–256. doi: 10.1038/283249a0. [DOI] [PubMed] [Google Scholar]
  18. Lee L. D., Baden H. P. Organisation of the polypeptide chains in mammalian keratin. Nature. 1976 Nov 25;264(5584):377–379. doi: 10.1038/264377a0. [DOI] [PubMed] [Google Scholar]
  19. Matoltsy A. G. Desmosomes, filaments, and keratohyaline granules: their role in the stabilization and keratinization of the epidermis. J Invest Dermatol. 1975 Jul;65(1):127–142. doi: 10.1111/1523-1747.ep12598093. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Osborn M., Franke W., Weber K. Direct demonstration of the presence of two immunologically distinct intermediate-sized filament systems in the same cell by double immunofluorescence microscopy. Vimentin and cytokeratin fibers in cultured epithelial cells. Exp Cell Res. 1980 Jan;125(1):37–46. doi: 10.1016/0014-4827(80)90186-x. [DOI] [PubMed] [Google Scholar]
  22. Shulman M., Wilde C. D., Köhler G. A better cell line for making hybridomas secreting specific antibodies. Nature. 1978 Nov 16;276(5685):269–270. doi: 10.1038/276269a0. [DOI] [PubMed] [Google Scholar]
  23. Skerrow D. The structure of prekeratin. Biochem Biophys Res Commun. 1974 Aug 19;59(4):1311–1316. doi: 10.1016/0006-291x(74)90457-4. [DOI] [PubMed] [Google Scholar]
  24. Steinert P. M. Structure of the three-chain unit of the bovine epidermal keratin filament. J Mol Biol. 1978 Jul 25;123(1):49–70. doi: 10.1016/0022-2836(78)90376-5. [DOI] [PubMed] [Google Scholar]
  25. Sun T. T., Green H. Immunofluorescent staining of keratin fibers in cultured cells. Cell. 1978 Jul;14(3):469–476. doi: 10.1016/0092-8674(78)90233-7. [DOI] [PubMed] [Google Scholar]
  26. Sun T. T., Green H. Keratin filaments of cultured human epidermal cells. Formation of intermolecular disulfide bonds during terminal differentiation. J Biol Chem. 1978 Mar 25;253(6):2053–2060. [PubMed] [Google Scholar]
  27. Sun T. T., Shih C., Green H. Keratin cytoskeletons in epithelial cells of internal organs. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2813–2817. doi: 10.1073/pnas.76.6.2813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Viac J., Staquet M. J., Thivolet J., Goujon C. Experimental production of antibodies against stratum corneum keratin polypeptides. Arch Dermatol Res. 1980;267(2):179–188. doi: 10.1007/BF00569104. [DOI] [PubMed] [Google Scholar]
  29. WALEN K. H., BROWN S. W. Chromosomes in a marsupial (Potorous tridactylis) tissue culture. Nature. 1962 Apr 28;194:406–406. doi: 10.1038/194406a0. [DOI] [PubMed] [Google Scholar]
  30. Winter H., Schweizer J., Goerttler K. Keratins as markers of malignancy in mouse epidermal tumors. Carcinogenesis. 1980 May;1(5):391–398. doi: 10.1093/carcin/1.5.391. [DOI] [PubMed] [Google Scholar]

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