Involucrin: A Constituent of Cross-Linked Envelopes and Marker of Squamous Maturation

  • Robert H. Rice
  • Scott M. Thacher


The coordinated program of terminal differentiation in epidermis yields a superficial layer of tough dead squames well suited to the protective function of the integument. These mature cells consist primarily of insoluble disulfide-bonded keratin tonofilaments, but also exhibit a “cornified envelope” immediately beneath the plasma membrane. This structure, consisting of protein, is resistant to keratinolytic agents (alkali, detergent and reducing agent) and organic solvents, but is sensitive to proteolytic digestion (Matoltsy and Balsamo 1955; Sun and Green 1976). The chemical stability of the envelope is attributable to a high degree of ε-(γ-glutamyl)lysine cross-linking arising from cellular transglutaminase activity (Sugawara 1977; Rice and Green 1977). Envelopes are conspicuous in hair and nail samples boiled in the presence of sodium dodecyl sulfate (SDS) and reducing agent (Rice and Green, unpublished), where their rigid interlocking convolutions contribute to the exceptional cohesiveness of the cells in these appendages (Green et al. 1982). Envelopes are present on the surface of all the stratified squamous epithelia of the human and have been observed in the cornified layer of epidermis of mammals, birds, reptiles and amphibians but not fish (Matoltsy 1977).


Stratify Squamous Epithelium Human Epidermis Squamous Differentiation Cornified Envelope Retinyl Acetate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Abernathy JL, Hill RL, Goldsmith LA (1977) e-(y-Glutamyl) lysine cross-links in human stratum corneum. J Biol Chem 252: 1837–1839Google Scholar
  2. Banks-Schlegel S, Green H (1980) Formation of epidermis by serially cultivated human epidermal cells transplanted as an epithelium to athymic mice. Transplantation 29: 308–313CrossRefPubMedGoogle Scholar
  3. Banks-Schlegel S, Green H (1981) Involucrin synthesis and tissue assembly by keratinocytes in natural and cultured human epithelia. J Cell Biol 90: 732–737CrossRefPubMedGoogle Scholar
  4. Buxman MM, Buehner GE, Wuepper KD (1976) Isolation of substrates of epidermal transglutaminase from bovine epidermis. Biochem Biophys Res Commun 73: 470–478CrossRefPubMedGoogle Scholar
  5. Buxman MM, Lobitz CJ, Wuepper KD (1980) Epidermal transglutaminase. Identification and purification of a soluble substrate with studies of in vitro cross-linking. J Biol Chem 255: 1200–1203Google Scholar
  6. Cline PR, Rice RH (1983) Modulation of involucrin and envelope competence in human keratinocytes by hydrocortisone, retinyl acetate and growth arrest. Cancer Res 43: 3203–3207PubMedGoogle Scholar
  7. Green H (1977) Terminal differentiation of cultured human epidermal cells. Cell 11: 405–415CrossRefPubMedGoogle Scholar
  8. Green H (1978) Cyclic AMP in relation to proliferation of the epidermal cell: a new view. Cell 15: 801–811CrossRefPubMedGoogle Scholar
  9. Green H, Fuchs E, Watt F (1982) Differentiated structural components of the keratinocyte. Organization of the cytoplasm. Cold Spring Harbor Symp Quant Biol 46: 293–301Google Scholar
  10. Heimann R, Rice RH (1983) Polycyclic aromatic hydrocarbon toxicity and induction of metabolism in cultivated esophageal and epidermal keratinocytes. Cancer Res 43: 4856–4862PubMedGoogle Scholar
  11. Kubilus J, Baden HP (1982) Isolation of two immunologically related transglutaminase sub- strates from cultured human keratinocytes. In Vitro 18: 447–455Google Scholar
  12. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685CrossRefPubMedGoogle Scholar
  13. Lechner JF, Haugen A, Autrup H, McClendon IA, Trump BF, Harris CC (1981) Clonal growth of epithelial cells from normal adult bronchus. Cancer Res 41: 2294–2304PubMedGoogle Scholar
  14. Lobitz CJ, Buxman MM (1982) Characterization and localization of bovine epidermal transglutaminase substrate. J Invest Dermatol 78: 150–154CrossRefPubMedGoogle Scholar
  15. Matoltsy AG (1977) The membrane of horny cells. In: Seiji M, Bernstein IA (eds) Biochemistry of cutaneous epidermal differentiation. Univ Tokyo Press, Tokyo, pp 93–109Google Scholar
  16. Matoltsy AG, Balsamo CA (1955) A study of the components of cornified epithelium of human skin. J Biophys Biochem Cytol 1: 339–360CrossRefPubMedGoogle Scholar
  17. Murphy GF, Flynn TC, Rice RH, Pinkus GS (1984) Involucrin expression in normal and neoplastic human skin: a marker for keratinocyte differentiation. J Invest Dermatol 82: 453–457CrossRefPubMedGoogle Scholar
  18. Phillips MA, Rice RH (1983) Convergent differentiation in cultured rat cells from nonkeratinized epithelia: keratinocyte character and intrinsic differences. J Cell Biol 97: 686–691CrossRefPubMedGoogle Scholar
  19. Rheinwald JG (1980) Serial cultivation of normal human epidermal keratinocytes. Methods Cell Biol 21: 229–254CrossRefGoogle Scholar
  20. Rheinwald JG, Green H (1975) Formation of a keratinizing epithelium in culture by a cloned line derived from a teratoma. Cell 6: 317–330CrossRefPubMedGoogle Scholar
  21. Rheinwald JG, Germain E, Beckett MA (1983) Expression of keratins and envelope proteins in normal and malignant human keratinocytes and mesothelial cells. In: Harris C, Autrup H (eds) Human carcinogenesis. Academic Press, London New York, pp 85–96Google Scholar
  22. Rice RH, Green H (1977) The cornified envelope of terminally differentiated human epidermal keratinocytes consists of cross-linked protein. Cell 11: 417–422CrossRefPubMedGoogle Scholar
  23. Rice RH, Green H (1978) Relationship of protein synthesis and transglutaminase activity to formation of the cross-linked envelope during terminal differentiation of the cultured human epidermal keratinocyte. J Cell Biol 76: 705–711CrossRefPubMedGoogle Scholar
  24. Rice RH, Green H (1979) Presence in human epidermal cells of a soluble protein precursor of the cross-linked envelope: activation of the cross-linking by calcium ions. Cell 18: 681–694CrossRefPubMedGoogle Scholar
  25. Rice RH, Pinkus GS, Warhol MJ, Antonioli DA (1984) Involucrin: biochemistry and immunohistochemistry. In: Sternberg S, DeLellis R (eds) Advances in immunohistochemistry. Masson, USA, pp 111–125Google Scholar
  26. Said JW, Nash G, Sassoon AF, Shintaku IP, Banks-Schlegel S (1983) Involucrin in lung tumors. A specific marker for squamous differentiation. Lab Invest 49: 563–568Google Scholar
  27. Simon M, Green H (1984) Participation of membrane-associated proteins in the formation of the cross-linked envelope of the keratinocyte. Cell 36: 327–334CrossRefGoogle Scholar
  28. Sugawara K (1977) Intermolecular cross-links in epidermal differentiation. In: Seiji M, Bernstein IA (eds) Biochemistry of cutaneous epidermal differentiation. Univ Tokyo Press, Tokyo, pp 387–397Google Scholar
  29. Sun TT, Green H (1976) Differentiation of the epidermal keratinocyte in cell culture: formation of the cornified envelope. Cell 9: 511–521CrossRefPubMedGoogle Scholar
  30. Towbin H, Staehelin T, Gorden J (1979) Electrophoretic transfer of proteins from acrylam_de gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354CrossRefPubMedGoogle Scholar
  31. Warhol MJ, Antonioli DA, Pinkus GS, Burke L, Rice RH (1982) Immunoperoxidase staining for involucrin: a potential diagnostic aid in cervicovaginal pathology. Hum Pathol 13: 1095–1099CrossRefPubMedGoogle Scholar
  32. Warhol MJ, Roth J, Lucocq JM, Pinkus GS, Rice RH (1985) Immuno-ultrastructural localization of involucrin in squamous epithelium and cultured keratinocytes. J Histochem Cytochem 33: 141–149CrossRefPubMedGoogle Scholar
  33. Watt FM, Green H (1981) Involucrin synthesis is correlated with cell size in human cultures. J Cell Biol 90: 738–742CrossRefPubMedGoogle Scholar
  34. Watt FM, Green H (1982) Stratification and terminal differentiation of cultured epidermal cells. Nature 295: 434–436CrossRefPubMedGoogle Scholar
  35. Weber K, Osborn M (1969) The reliability of molecular weight estimates for proteins in polyacrylamide gel electrophoresis. J Biol Chem 244: 4406–4412PubMedGoogle Scholar
  36. Zettergren JG, Peterson LL, Wuepper KD (1984) Keratolinin: The soluble substrate of epidermal transglutaminase from human and bovine tissue. Proc Natl Acad Sci USA 81: 238–242Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1986

Authors and Affiliations

  • Robert H. Rice
    • 1
  • Scott M. Thacher
    • 1
  1. 1.Charles A. Dana Laboratory of ToxicologyHarvard School of Public HealthBostonUSA

Personalised recommendations