Skip to main content
SpringerLink
Log in

Identification of the conserved, conformation-dependent cytokeratin epitope recognized by monoclonal antibody (lu-5)

  • Published:
Virchows Archiv A Aims and scope Submit manuscript

Summary

The epitope recognized by the murine monoclonal antibody (mAB lu-5) recently described as a formaldehyde-resistant, “pan-epithelial marker” of great value in tumour diagnosis is located on the surface of cytokeratin filaments. It has been preserved during vertebrate evolution from amphibia to man. As this epitope is not reactive after SDS-polyacrylamide gel electrophoresis (SDS-PAGE), the epitope-bearing protein has been identified by a dot-blot antibody binding assay, using purified proteins in which the epitope is reconstituted. We show that the epitope is present in most cytokeratin polypeptides of both the acidic (type I) and basic (type II) subfamily but does not occur in other cytoskeletal proteins. The location of this widespread epitope is discussed with respect to homologies of amino acid sequences of cytokeratins and their conformations.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Achtstätter T, Hatzfeld M, Quinlan RA, Parmelee DC, Franke WW (1986) Separation of cytokeratin polypeptides by gel electrophoretic and chromatographic techniques and their identification by immunoblotting. Meth Enzymol 134:355–371

    Google Scholar 

  • Bader BL, Magin TM, Hatzfeld M, Franke WW (1986) Amino acid sequence and gene organization of cytokeratin no. 19, an exceptional tail-less intermediate filament protein. EMBO J 5:1865–1875

    Google Scholar 

  • Bannasch P, Zerban H, Schmid E, Franke WW (1980) Liver tumors distinguished by immunofluorescence microscopy with antibodies to proteins of intermediate-sized filaments. Proc Natl Acad Sci USA 77:4948–4952

    Google Scholar 

  • Barlow DJ, Edwards MS, Thornton JM (1986) Continuous and discontinuous protein antigenic determinants. Nature 322:747–748

    Google Scholar 

  • Bartek J, Durban EM, Hallowes RC, Taylor-Papadimitriou J (1985) A subclass of luminal epithelial cells in the human mammary gland, defined by antibodies to cytokeratins. J Cell Sci 75:17–33

    Google Scholar 

  • Bartnik E, Osborn M, Weber K (1985) IFs in non-neuronal cells of invertebrates: isolation and biochemical characterization of keratin-like intermediate filaments from the esophageal epithelium of the molluscHelix pomatia. J Cell Biol 101:427–440

    Google Scholar 

  • Bartnik E, Osborn M, Weber K (1986) Intermediate filaments in muscle and epithelial cells of nematodes. J Cell Biol 102:2033–2041

    Google Scholar 

  • Battifora H, Sun TT, Rao S, Bahu R (1980) Antikeratin antibodies in tumor diagnosis: thymoma versus lymphoma. Hum Pathol 11:635–641

    Google Scholar 

  • Benavente R, Krohne G, Franke WW (1985) Cell type-specific expression of nuclear lamina proteins during development ofXenopus laevis. Cell 41:177–190

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Google Scholar 

  • Cooper D, Schermer A, Pruss R, Sun T-T (1984) The use of aIF, AE1, and AE3 monoclonal antibodies for the identification and classification of mammalian epithelial keratins. Differentiation 28:30–35

    Google Scholar 

  • Cooper D, Schermer A, Sun T-T (1985) Classification of human epithelia and their neoplasms using monoclonal antibodies to keratins: Strategies, applications, and limitations. Lab Invest 52:243–256

    Google Scholar 

  • Corson JM (1986) Keratin protein immunohistochemistry in surgical pathology practice. In: Sommers SC, Rosen PP, Fechner RE (eds) Pathology Annual, vol 21, part 2. Appleton-Century-Crofts, Norwalk, Connecticut, pp 49–81

    Google Scholar 

  • Cowin P, Kapprell H-P, Franke WW (1985) The complement of desmosomal plaque proteins in different cell types. J Cell Biol 101:1442–1454

    Google Scholar 

  • Cowin P, Kapprell H-P, Franke WW, Tamkun J, Hynes RO (1986) Plakoglobin: A protein common to different kinds of intercellular adhering junctions. Cell 46:1063–1073

    Google Scholar 

  • Crewther WG, Dowling LM, Steinert PM, Parry DAD (1983) Structure of intermediate filaments. Int J Biol Macromol 5:267–274

    Google Scholar 

  • Debus E, Weber K, Osborn M (1982) Monoclonal cytokeratin antibodies that distinguish simple from stratified squamous epithelia: Characterization of human tissues. EMBO J 1:1641–1647

    Google Scholar 

  • Debus E, Moll R, Franke WW, Weber K, Osborn M (1984) Immunohistochemical distinction of human carcinomas by cytokeratin typing with monoctonal antibodies. Am J Pathol 114:121–130

    Google Scholar 

  • Dowling LM, Crewther WG, Inglis AS (1986) The primary structure of component 8c-1, a subunit protein of intermediate filaments in wool keratin. Biochem J 236:695–703

    Google Scholar 

  • Franke WW, Schmid E, Osborn M, Weber K (1978a) Different intermediate-sized filaments distinguished by immunofluorescence microscopy. Proc Natl Acad Sci USA 75:5034–5038

    Google Scholar 

  • Franke WW, Weber K, Osborn M, Schmid E, Freudenstein C (1978b) Antibody to prekeratin. Decoration of tonofilament-like arrays in various cells of epithelial character. Exp Cell Res 116:429–445

    Google Scholar 

  • Franke WW, Appelhans B, Schmid E, Freudenstein C, Osborn M, Weber K (1979a) Identification and characterization of epithelial cells in mammalian tissues by immunofluorescence microscopy using antibodies to prekeratin. Differentiation 15:7–25

    Google Scholar 

  • Franke WW, Schmid E, Osborn M (1979b) HeLa cells contain intermediate-sized filaments of the prekeratin type. Exp Cell Res 118:95–109

    Google Scholar 

  • Franke WW, Schmid E, Winter S, Osborn M, Weber K (1979c) Widespread occurrence of intermediate-sized filaments of the vimentin-type in cultured cells from diverse vertebrates. Exp Cell Res 123:25–46

    Google Scholar 

  • Franke WW, Schmid E, Freudenstein C, Appelhans B, Osborn M, Weber K, Keenan TW (1980) Intermediate-sized filaments of the prekeratin type in myoepithelial cells. J Cell Biol 84:633–654

    Google Scholar 

  • Franke WW, Mayer D, Schmid E, Denk H, Borenfreund E (1981a) Differences of expression of cytoskeletal proteins in cultured rat hepatocytes and hepatoma cells. Exp Cell Res 134:345–365

    Google Scholar 

  • Franke WW, Schiller DL, Moll R, Winter S, Schmid E, Engelbrecht I, Denk H (1981b) Diversity of cytokeratins. Differentiation specific expression of cytokeratin polypeptides in epithelial cells and tissues. J Mol Biol 153:933–959

    Google Scholar 

  • Franke WW, Schmid E, Schiller DL, Winter S, Jarasch E-D, Moll R, Denk H, Jackson B, Illmensee K (1982) Differentiation-related patterns of expression of proteins of intermediate-sized filaments in tissues and cultured cells. Cold Spring Harbor Symp Quant Biol 46:431–453

    Google Scholar 

  • Franke WW, Schiller DL, Hatzfeld M, Winter S (1983) Protein complexes of intermediate-sized filaments: Melting of cytokeratin complexes in urea reveals different polypeptide separation characteristics. Proc Natl Acad Sci USA 80:7113–7117

    Google Scholar 

  • Franke WW, Schmid E, Mittnacht S, Grund C, Jorcano JL (1984) Integration of different keratins into the same filament system after microinjection of mRNA for epidermal keratins into kidney epithelial cells. Cell 36:813–825

    Google Scholar 

  • Franz JK, Gall L, Williams MA, Picheral B, Franke WW (1983) Intermediate-size filaments in a germ cell: expression of cytokeratins in oocytes and eggs of the frogXenopus. Proc Natl Acad Sci USA 80:6254–6258

    Google Scholar 

  • Franz JK, Franke WW (1986) Cloning of cDNA and amino acid sequence of a cytokeratin expressed in oocytes ofXenopus laevis. Proc Natl Acad Sci USA 83:6475–6479

    Google Scholar 

  • Fuchs EV, Coppock SM, Green FH, Cleveland DW (1981) Two distinct classes of keratin genes and their evolutionary significance. Cell 27:75–84

    Google Scholar 

  • Fuchs E, Green H (1981) Regulation of terminal differentiation of cultured human keratinocytes by vitamin A. Cell 25:617–625

    Google Scholar 

  • Fuchs E, Hanukoglu I, Marchuk D, Grace MP, Kim KH (1985) The nature and significance of differential keratin gene expression. In: Wang E, Fischman D, Liem RKH, Sun T-T (eds) Intermediate Filaments, vol 455. The New York Academy of Sciences, New York, pp 436–450

    Google Scholar 

  • Gabbiani G, Kapanci Y, Barazzone P, Franke WW (1981) Immunochemical identification of intermediate-sized filaments of in human neoplastic cells: A diagnostic aid for the surgical pathologist. Am J Pathol 104:206–216

    Google Scholar 

  • Geisler N, Weber K (1980) Purification of smooth-muscle desmin and a protein-chemical comparison of desmins from chicken gizzard and hog stomach. Eur J Biochem 111:425–433

    Google Scholar 

  • Geisler N, Weber K (1981) Isolation of polymerization-competent vimentin from porcine eye lens tissue. FEBS Lett 125:253–256

    Google Scholar 

  • Geisler N, Weber K (1982) The amino acid sequence of chicken muscle desmin provides a common structural model for intermediate filament proteins. EMBO J 1:1649–1656

    Google Scholar 

  • Gigi O, Geiger B, Eshhar Z, Moll R, Schmid E, Winter S, Schiller DL, Franke WW (1982) Detection of a cytokeratin determinant common to diverse epithelial cells by a broadly cross-reacting monoclonal antibody. EMBO J 1:1429–1437

    Google Scholar 

  • Gown AM, Vogel AM (1984) Monoclonal antibodies to human intermediate filament proteins. II. Distribution of filament proteins in normal human tissues. Am J Pathol 114:309

    Google Scholar 

  • Hanukoglu I, Fuchs E (1983) The cDNA sequence of a type II cytoskeletal keratin reveals constant and variable structural domains among keratins. Cell 33:915–924

    Google Scholar 

  • Hatzfeld M, Franke WW (1985) Pair formation and promiscuity of cytokeratins: Formation in vitro of heterotypic complexes and intermediate-sized filaments by homologous and heterologous recombinations of purified polypeptides. J Cell Biol 101:1826–1841

    Google Scholar 

  • Hazan R, Denk H, Franke WW, Lackinger E, Schiller DL (1986) Change of cytokeratin organization during development of mallory bodies as revealed by a monoclonal antibody. Lab Invest 54:543–553

    Google Scholar 

  • Heid HW, Werner W, Franke WW (1986) The complement of native α-keratin polypeptides of hair-forming cells: A subset of eight polypeptides that differ from epithelial cytokeratins. Differentiation 31:141–153

    Google Scholar 

  • Hoffmann W, Frank JK, Franke WW (1985) Amino acid sequence microheterogeneities of basic (type II) cytokeratins ofXenopus laevis epidermis and evolutionary conservativity of helical and non-helical domains. J Mol Biol 184:713–724

    Google Scholar 

  • Holthöfer H, Miettinen A, Paasivuo R, Lehto V-P, Linder E, Alfthan O, Virtanen I (1983) Cellular origin and differentiation of renal carcinomas. Lab Invest 49:317–326

    Google Scholar 

  • Jonas E, Sargent TD, Dawid IB (1985) Epidermal keratin gene expressed in embryos ofXenopus laevis. Proc Natl Acad Sci USA 82:5413–5417

    Google Scholar 

  • Knapp AC, Franke WW, Heid H, Hatzfeld M, Jorcano JL, Moll R (1986) Cytokeratin no. 9, an epidermal type I keratin characteristic of a special program of keratinocyte differentiation displaying body site specificity. J Cell Biol 103:657–667

    Google Scholar 

  • Lane EB (1982) Monoclonal antibodies provide specific intramolecular markers for the study of epithelial tonofilament organization. J Cell Biol 92:665–673

    Google Scholar 

  • Lane EB, Bártek J, Purkis PE, Leigh IM (1985) Keratin antigens in differentiating skin. In: Wang E, Fischman D, Liem RKH, Sun T-T (eds) Intermediate Filaments, vol 455. The New York Academy of Sciences, New York, pp 241–258

    Google Scholar 

  • Leube RE, Bosch FX, Romano V, Zimbelmann R, Höfler H, Franke WW (1986) Cytokeratin expression in simple epithelia. III. Detection of mRNAs encoding human cytokeratins nos. 8 and 18 in normal and tumor cells by hybridization with cDNA sequences in vitro and in situ. Differentiation 33:69–85

    Google Scholar 

  • Magin TM, Jorcano JL, Franke WW (1986). Cytokeratin expression in simple epithelia. II. cDNA cloning and sequence characteristics of bovine cytokeratin A (no. 8). Differentiation 30:254–264

    Google Scholar 

  • Makin CA, Bobrow LG, Bodmer WF (1984) Monoclonal antibody to cytokeratin for use in routine histopathology. J Clin Pathol 37:975–983

    Google Scholar 

  • Marchuk D, McCrohon S, Fuchs E (1984) Complete sequence of a gene encoding a human type I keratin: Sequences homologous to enhancer elements in the regulatory region of the gene. Cell 39:491–498

    Google Scholar 

  • Miettinen M, Lehto V-P, Virtanen I (1984) Antibodies to intermediate filament proteins in the diagnosis and classification of human tumors. Ultrastruct Pathol 7:83

    Google Scholar 

  • Moll R, Franke WW, Schiller DL, Geiger B, Krepler R (1982) the catalog of human cytokeratin polypeptides: Patterns of expression of specific cytokeratins in normal epithelia, tumors and cultured cells. Cell 31:11–24

    Google Scholar 

  • Moll R, Cowin P, Kapprell H-P, Franke WW (1986) Desmosomal proteins: New Markers for identification and classification of tumors. Lab Invest 54:4–25

    Google Scholar 

  • Osborn M, Weber K (1983) Tumor diagnosis by intermediate filament typing: A novel tool for surgical pathology. Lab Invest 48:372–394

    Google Scholar 

  • Oshima RG, Millan JL, Ceceña G (1986) Comparison of mouse and human keratin 18: A component of intermediate filaments expressed prior to implantation. Differentiation 33:61–68

    Google Scholar 

  • Overbeck von J, Stähli C, Gudat F, Carmann H, Lautenschlager C, Dürmüller U, Takacs B, Miggiano V, Staehelin Th, Heitz PhU (1985) Immunohistochemical characterization of an anti-epithelial monoclonal antibody (mAB lu-5) Virchows Arch A (Pathol Anat) 407:1–12

    Google Scholar 

  • Pruss RM, Mirsky R, Ruff MC, Thorpe R, Dowding AJ, Anderton BH (1981) All classes of intermediate filaments share a common antigenic determinant defined by a monoclonal antibody. Cell 27:419–428

    Google Scholar 

  • Quinlan RA, Franke WW (1983) Molecular interactions in intermediate-sized filaments revealed by chemical crosslinking. Heteropolymers of vimentin and glial filament protein in cultured human glioma cells. Eur J Biochem 132:477–484

    Google Scholar 

  • Quinlan RA, Cohlberg JA, Schiller DL, Hatzfeld M, Franke WW (1984) Heterotypic tetramer (A2D2) complexes of non-epidermal keratins isolated from cytoskeletons of rat hepatocytes and hepatoma cells. J Mol Biol 178:365–388

    Google Scholar 

  • Quinlan RA, Hatzfeld M, Franke WW, Lustig A, Schulthess T, Engel J (1986) Characterization of dimer subunits of intermediate filament proteins. J Mol Biol 192:337–349

    Google Scholar 

  • Ramaekers F, Huysmans A, Moesker O, Kant A, Jap P, Herman C, Vooijs P (1983a) Monoclonal antibody to keratin filaments, specific for glandular epithelia and their tumors. Lab Invest 49:353–361

    Google Scholar 

  • Ramaekers FCS, Puts JJG, Moesker O, Kant A, Huysmans A, Haag D, Jap PHK, Herman CJ, Vooijs GP (1983b) Antibodies to intermediate filament proteins in the immunohistochemical identification of human tumours: an overview. Histochem J 15:691–713

    Google Scholar 

  • Roop DR, Cheng CK, Titterington L, Meyers CA, Stanley JR, Steinert PM, Yuspa SH (1984) Synthetic peptides corresponding to keratin subunits elicit highly specific antibodies. J Biol Chem 259:8037–8040

    Google Scholar 

  • Schiller DL, Franke WW, Geiger B (1982) A subfamily of relatively large and basic cytokeratin polypeptides as defined by peptide mapping is represented by one or several polypeptides in epithelial cells. EMBO J 1:761–769

    Google Scholar 

  • Schlegel R, Banks-Schlegel S, McLeod JA, Pinkus GS (1980) Immunoperoxidase localization of keratin in human neoplasms. Am J Pathol 101:41–50

    Google Scholar 

  • Schmid E, Tapscott S, Bennett GS, Croop J, Fellini SA, Holtzer H, Franke WW (1979) Differential location of different types of intermediate-sized filaments in various tissues of the chick embryo. Differentiation 15:27–40

    Google Scholar 

  • Schmid E, Schiller DL, Grund C, Stadler J, Franke WW (1983) Tissue type specific expression in a cultured epithelial cell line from bovine mammary gland. J Cell Biol 96:37–50

    Google Scholar 

  • Söllner P, Quinlan RA, Franke WW (1985) Identification of a distinct soluble subunit of an intermediate filament protein: Tetrameric vimentin from living cells. Proc Natl Acad Sci USA 82:7929–7933

    Google Scholar 

  • Spagnolo DV, Michie SA, Crabtree GS, Warnke RA, Rouse RV (1985) Monoclonal anti-keratin (AE1) reactivity in routinely processed tissue from 166 human neoplasms. AM J Clin Pathol 84:697–704

    Google Scholar 

  • Steinert PM, Parry DAD, Idler WW, Johnson LD, Steven AC, Roop DR (1985a) Amino acid sequences of mouse and human epidermal type II keratins of Mr 67,000 provide a systematic basis for the structural and functional diversity of the end domains of keratin intermediate filament subunits. J Biol Chem 260:7142–7149

    Google Scholar 

  • Steinert PM, Steven AC, Roop DR (1985b) The molecular biology of intermediate filaments. Cell 42:411–419

    Google Scholar 

  • Sun T-T, Green H (1978) Immunofluorescent staining of keratin fibers in cultured cells. Cell 14:469–476

    Google Scholar 

  • Sun T-T, Shih CH, Green H (1979) Keratin cytoskeletons in epithelial cells of internal organs. Proc Natl Acad Sci USA 76:2813–2817

    Google Scholar 

  • Sun T-T, Eichner R, Nelson WG, Tseng SCG, Weiss RA, Jarvinen M, Woodcock-Mitchell J (1983) Keratin classes: molecular markers for different types of epithelial differentiation. J Invest Dermatol 81:109s-115s

    Google Scholar 

  • Sun T-T, Eichner R, Schermer A, Cooper D, Nelson WG, Weiss RA (1984) Classification, expression, and possible mechanisms of evolution of mammalian epithelial keratins: a unifying model. In: Levine AJ, Vande Woude GF, Topp WC, Watson JD (eds) Cancer Cells 1, The transformed phenotype. Cold Spring Harbor Laboratory, Cold Spring Harbor

    Google Scholar 

  • Tseng SCG, Jarvinen MJ, Nelson WG, Huang J-W, Woodcock-Mitchell J, Sun T-T (1982) Correlation of specific keratins with different types of epithelial differentiation: monoclonal antibody studies. Cell 30:361–372

    Google Scholar 

  • Van Muijen GNP, Ruiter DJ, Franke WW, Achtstätter T, Haasnoot WHB, Ponec M, Warnaar SO (1986) Cell type heterogeneity of cytokeratin expression in complex epithelia and carcinomas demonstrated by monoclonal antibodies specific for cytokeratins nos. 4 and 13. Exp Cell Res 162:97–113

    Google Scholar 

  • Viac J, Reano A, Brochier J, Staquet M-J, Thivolet J (1983) Reactivity pattern of a monoclonal antikeratin antibody (KL1). J Invest Dermatol 81:351–354

    Google Scholar 

  • Walter MF, Biessmann H (1984) Intermediate-sized filaments in Drosophila tissue culture cells. J Cell Biol 99:1468–1477

    Google Scholar 

  • Weber K, Osborn M, Franke WW (1980) Antibodies against merokeratin from sheep wool decorate cytokeratin filaments in non-keratinizing epithelial cells. Eur J Cell Biol 23:110–114

    Google Scholar 

  • Weber K, Geisler N (1984) Intermediate filaments from wool α-keratin to neurofilaments: a structural overview. In: Levine AJ, Vande Woude GF, Topp WC, Watson JD (eds) Cancer Cells 1, The transformed phenotype. Cold Spring Harbor Laboratory, Cold Spring Harbor, pp 153–159

    Google Scholar 

  • Woods EF (1983) The number of polypeptide chains in the rod domain of bovine epidermal keratin. Biochem Int 7:769–774

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Membrane Biology and Biochemistry, Institute of Cell and Tumor Biology, German Cancer Research Center, Im Neuenheimer Feld 280, D-6900, Heidelberg, Federal Republic of Germany

    W. W. Franke & S. Winter

  2. Department of Pathology, University of Basel, P.O. Box, CH-4003, Basel, Switzerland

    J. von Overbeck, F. Gudat & P. U. Heitz

  3. Central Research Department, F. Hoffman-La Roche Co, Basel, Switzerland

    C. Stähli

Authors
  1. W. W. Franke
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S. Winter
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. von Overbeck
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. F. Gudat
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. P. U. Heitz
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. C. Stähli
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Franke, W.W., Winter, S., von Overbeck, J. et al. Identification of the conserved, conformation-dependent cytokeratin epitope recognized by monoclonal antibody (lu-5). Vichows Archiv A Pathol Anat 411, 137–147 (1987). https://doi.org/10.1007/BF00712737

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00712737

Key words

Search

Navigation