Amino Acids

, Volume 49, Issue 3, pp 635–642 | Cite as

Characterization of TG2 and TG1–TG2 double knock-out mouse epidermis

  • Consuelo Pitolli
  • Valentina Pietroni
  • Lyuben Marekov
  • Alessandro Terrinoni
  • Kiyofumi Yamanishi
  • Cinzia Mazzanti
  • Gerry Melino
  • Eleonora Candi
Original Article

Abstract

Transglutaminases (TGs) are a family of enzymes that catalyse the formation of isopeptide bonds between the γ-carboxamide groups of glutamine residues and the ε-amino groups of lysine residues leading to cross-linking reactions among proteins. Four members, TG1, TG2, TG3, and TG5, of the nine mammalian enzymes are expressed in the skin. TG1, TG3 and TG5 crosslinking properties are fundamental for cornified envelope assembly. In contrast, the role of TG2 in keratinization has never been studied at biochemical level in vivo. In this study, taking advantage of the TG2 knock-out (KO) and TG1 heterozygous mice, we generated and characterized the epidermis of TG1–TG2 double knock-out (DKO) mice. We performed morphological analysis of the epidermis and evaluation of the expression of differentiation markers. In addition, we performed analysis of the amino acid composition from isolated corneocytes. We found a significant change in amino acid composition in TG1KO cornified cell envelopes (CEs) while TG2KO amino acid composition was similar to wild-type CEs. Our results confirm a key role of TG1 in skin differentiation and CE assembly and demonstrate that TG2 is not essential for CE assembly and skin formation.

Keywords

Keratinocyte differentiation Cornified cell envelope Transglutaminase 

Abbreviations

TGs

Transglutaminases

ECM

Extracellular matrix

KO

Knock-out mouse

DKO

Double knock-out mouse

CE

Cornified cell envelope

References

  1. Aeschlimann D, Paulsson M (1991) Cross-linking of laminin-nidogen complexes by tissue transglutaminase. A novel mechanism for basement membrane stabilization. J Biol Chem 266(23):15308–15317PubMedGoogle Scholar
  2. Candi E, Melino G, Mei G, Tarcsa E, Chung SI, Marekov LN, Steinert PM (1995) Biochemical, structural, and transglutaminase substrate properties of human loricrin, the major epidermal cornified cell envelope protein. J Biol Chem 270(44):26382–26390CrossRefPubMedGoogle Scholar
  3. Candi E, Melino G, Lahm A, Ceci R, Rossi A, Kim IG, Ciani B, Steinert PM (1998) Transglutaminase 1 mutations in lamellar ichthyosis. Loss of activity due to failure of activation by proteolytic processing. J Biol Chem 273(22):13693–13702CrossRefPubMedGoogle Scholar
  4. Candi E, Tarcsa E, Idler WW, Kartasova T, Marekov LN, Steinert PM (1999) Transglutaminase cross-linking properties of the small proline-rich 1 family of cornified cell envelope proteins. Integration with loricrin. J Biol Chem 274(11):7226–7237CrossRefPubMedGoogle Scholar
  5. Candi E, Oddi S, Terrinoni A, Paradisi A, Ranalli M, Finazzi-Agro A, Melino G (2001) Transglutaminase 5 cross-links loricrin, involucrin, and small proline-rich proteins in vitro. J Biol Chem 276(37):35014–35023. doi:10.1074/jbc.M010157200 CrossRefPubMedGoogle Scholar
  6. Candi E, Oddi S, Paradisi A, Terrinoni A, Ranalli M, Teofoli P, Citro G, Scarpato S, Puddu P, Melino G (2002) Expression of transglutaminase 5 in normal and pathologic human epidermis. J Invest Dermatol 119(3):670–677. doi:10.1046/j.1523-1747.2002.01853.x CrossRefPubMedGoogle Scholar
  7. Candi E, Schmidt R, Melino G (2005) The cornified envelope: a model of cell death in the skin. Nat Rev Mol Cell Biol 6(4):328–340. doi:10.1038/nrm1619 CrossRefPubMedGoogle Scholar
  8. Craiglow BG (2013) Ichthyosis in the newborn. Semin Perinatol 37(1):26–31. doi:10.1053/j.semperi.2012.11.001 CrossRefPubMedPubMedCentralGoogle Scholar
  9. De Laurenzi V, Melino G (2001) Gene disruption of tissue transglutaminase. Mol Cell Biol 21(1):148–155. doi:10.1128/MCB.21.1.148-155.2001 CrossRefPubMedPubMedCentralGoogle Scholar
  10. Dolynchuk KN, Bendor-Samuel R, Bowness JM (1994) Effect of putrescine on tissue transglutaminase activity in wounds: decreased breaking strength and increased matrix fucoprotein solubility. Plast Reconstr Surg 93(3):567–573CrossRefPubMedGoogle Scholar
  11. Etoh Y, Simon M, Green H (1986) Involucrin acts as a transglutaminase substrate at multiple sites. Biochem Biophys Res Commun 136(1):51–56CrossRefPubMedGoogle Scholar
  12. Fischer J (2009) Autosomal recessive congenital ichthyosis. J Invest Dermatol 129(6):1319–1321. doi:10.1038/jid.2009.57 CrossRefPubMedGoogle Scholar
  13. Gentile V, Thomazy V, Piacentini M, Fesus L, Davies PJ (1992) Expression of tissue transglutaminase in Balb-C 3T3 fibroblasts: effects on cellular morphology and adhesion. J Cell Biol 119(2):463–474CrossRefPubMedGoogle Scholar
  14. Greenberg CS, Birckbichler PJ, Rice RH (1991) Transglutaminases: multifunctional cross-linking enzymes that stabilize tissues. FASEB J 5(15):3071–3077PubMedGoogle Scholar
  15. Griffin M, Casadio R, Bergamini CM (2002) Transglutaminases: nature’s biological glues. Biochem J 368(Pt 2):377–396. doi:10.1042/BJ20021234 CrossRefPubMedPubMedCentralGoogle Scholar
  16. Haroon ZA, Hettasch JM, Lai TS, Dewhirst MW, Greenberg CS (1999) Tissue transglutaminase is expressed, active, and directly involved in rat dermal wound healing and angiogenesis. FASEB J 13(13):1787–1795PubMedGoogle Scholar
  17. Huber M, Rettler I, Bernasconi K, Frenk E, Lavrijsen SP, Ponec M, Bon A, Lautenschlager S, Schorderet DF, Hohl D (1995) Mutations of keratinocyte transglutaminase in lamellar ichthyosis. Science 267(5197):525–528CrossRefPubMedGoogle Scholar
  18. Jarnik M, Kartasova T, Steinert PM, Lichti U, Steven AC (1996) Differential expression and cell envelope incorporation of small proline-rich protein 1 in different cornified epithelia. J Cell Sci 109(Pt 6):1381–1391PubMedGoogle Scholar
  19. Kalinin A, Marekov LN, Steinert PM (2001) Assembly of the epidermal cornified cell envelope. J Cell Sci 114(Pt 17):3069–3070PubMedGoogle Scholar
  20. Kalinin AE, Kajava AV, Steinert PM (2002) Epithelial barrier function: assembly and structural features of the cornified cell envelope. BioEssays 24(9):789–800. doi:10.1002/bies.10144 CrossRefPubMedGoogle Scholar
  21. Kuramoto N, Takizawa T, Takizawa T, Matsuki M, Morioka H, Robinson JM, Yamanishi K (2002) Development of ichthyosiform skin compensates for defective permeability barrier function in mice lacking transglutaminase 1. J Clin Invest 109(2):243–250. doi:10.1172/JCI13563 CrossRefPubMedPubMedCentralGoogle Scholar
  22. Lorand L, Graham RM (2003) Transglutaminases: crosslinking enzymes with pleiotropic functions. Nat Rev Mol Cell Biol 4(2):140–156. doi:10.1038/nrm1014 CrossRefPubMedGoogle Scholar
  23. Matsuki M, Yamashita F, Ishida-Yamamoto A, Yamada K, Kinoshita C, Fushiki S, Ueda E, Morishima Y, Tabata K, Yasuno H, Hashida M, Iizuka H, Ikawa M, Okabe M, Kondoh G, Kinoshita T, Takeda J, Yamanishi K (1998) Defective stratum corneum and early neonatal death in mice lacking the gene for transglutaminase 1 (keratinocyte transglutaminase). Proc Natl Acad Sci USA 95(3):1044–1049CrossRefPubMedPubMedCentralGoogle Scholar
  24. Michel S, Schmidt R, Shroot B, Reichert U (1988) Morphological and biochemical characterization of the cornified envelopes from human epidermal keratinocytes of different origin. J Invest Dermatol 91(1):11–15CrossRefPubMedGoogle Scholar
  25. Nanda N, Iismaa SE, Owens WA, Husain A, Mackay F, Graham RM (2001) Targeted inactivation of Gh/tissue transglutaminase II. J Biol Chem 276(23):20673–20678. doi:10.1074/jbc.M010846200 CrossRefPubMedGoogle Scholar
  26. Nemes Z, Steinert PM (1999) Bricks and mortar of the epidermal barrier. Exp Mol Med 31(1):5–19. doi:10.1038/emm.1999.2 CrossRefPubMedGoogle Scholar
  27. Nemes Z, Marekov LN, Fesus L, Steinert PM (1999a) A novel function for transglutaminase 1: attachment of long-chain omega-hydroxyceramides to involucrin by ester bond formation. Proc Natl Acad Sci USA 96(15):8402–8407CrossRefPubMedPubMedCentralGoogle Scholar
  28. Nemes Z, Marekov LN, Steinert PM (1999b) Involucrin cross-linking by transglutaminase 1. Binding to membranes directs residue specificity. J Biol Chem 274(16):11013–11021CrossRefPubMedGoogle Scholar
  29. Raghunath M, Hopfner B, Aeschlimann D, Luthi U, Meuli M, Altermatt S, Gobet R, Bruckner-Tuderman L, Steinmann B (1996) Cross-linking of the dermo-epidermal junction of skin regenerating from keratinocyte autografts. Anchoring fibrils are a target for tissue transglutaminase. J Clin Invest 98(5):1174–1184. doi:10.1172/JCI118901 CrossRefPubMedPubMedCentralGoogle Scholar
  30. Reichert U, Michel S, Schmidt C (2012) The cornified envelope: a key structure of terminally differentiating keratinocytes. In: Darmon M, Blumenberg M (eds) Molecular biology of the skin: the keratinocyte, vol 1. Academic Press, San Diego, pp 107–146Google Scholar
  31. Rice RH, Green H (1977) The cornified envelope of terminally differentiated human epidermal keratinocytes consists of cross-linked protein. Cell 11(2):417–422CrossRefPubMedGoogle Scholar
  32. Rorke EA, Eckert RL (1991) Stable expression of transfected human involucrin gene in various cell types: evidence for in situ cross-linking by type I and type II transglutaminase. J Invest Dermatol 97(3):543–548CrossRefPubMedGoogle Scholar
  33. Russell LJ, DiGiovanna JJ, Rogers GR, Steinert PM, Hashem N, Compton JG, Bale SJ (1995) Mutations in the gene for transglutaminase 1 in autosomal recessive lamellar ichthyosis. Nat Genet 9(3):279–283. doi:10.1038/ng0395-279 CrossRefPubMedGoogle Scholar
  34. Sandler B, Hashimoto K (1998) Collodion baby and lamellar ichthyosis. J Cutan Pathol 25(2):116–121CrossRefPubMedGoogle Scholar
  35. Simon M, Green H (1988) The glutamine residues reactive in transglutaminase-catalyzed cross-linking of involucrin. J Biol Chem 263(34):18093–18098PubMedGoogle Scholar
  36. Steinert PM (2000) The complexity and redundancy of epithelial barrier function. J Cell Biol 151(2):F5–F8CrossRefPubMedPubMedCentralGoogle Scholar
  37. Steinert PM, Marekov LN (1995) The proteins elafin, filaggrin, keratin intermediate filaments, loricrin, and small proline-rich proteins 1 and 2 are isodipeptide cross-linked components of the human epidermal cornified cell envelope. J Biol Chem 270(30):17702–17711CrossRefPubMedGoogle Scholar
  38. Steinert PM, Marekov LN (1997) Direct evidence that involucrin is a major early isopeptide cross-linked component of the keratinocyte cornified cell envelope. J Biol Chem 272(3):2021–2030CrossRefPubMedGoogle Scholar
  39. Steinert PM, Chung SI, Kim SY (1996) Inactive zymogen and highly active proteolytically processed membrane-bound forms of the transglutaminase 1 enzyme in human epidermal keratinocytes. Biochem Biophys Res Commun 221(1):101–106. doi:10.1006/bbrc.1996.0552 CrossRefPubMedGoogle Scholar
  40. Stephens P, Grenard P, Aeschlimann P, Langley M, Blain E, Errington R, Kipling D, Thomas D, Aeschlimann D (2004) Crosslinking and G-protein functions of transglutaminase 2 contribute differentially to fibroblast wound healing responses. J Cell Sci 117(Pt 15):3389–3403. doi:10.1242/jcs.01188 CrossRefPubMedGoogle Scholar
  41. Steven AC, Steinert PM (1994) Protein composition of cornified cell envelopes of epidermal keratinocytes. J Cell Sci 107(Pt 2):693–700PubMedGoogle Scholar
  42. Steven AC, Bisher ME, Roop DR, Steinert PM (1990) Biosynthetic pathways of filaggrin and loricrin—two major proteins expressed by terminally differentiated epidermal keratinocytes. J Struct Biol 104(1–3):150–162CrossRefPubMedGoogle Scholar
  43. Takahashi M, Tezuka T, Kakegawa H, Katunuma N (1994) Linkage between phosphorylated cystatin alpha and filaggrin by epidermal transglutaminase as a model of cornified envelope and inhibition of cathepsin L activity by cornified envelope and the conjugated cystatin alpha. FEBS Lett 340(3):173–176CrossRefPubMedGoogle Scholar
  44. Tarcsa E, Candi E, Kartasova T, Idler WW, Marekov LN, Steinert PM (1998) Structural and transglutaminase substrate properties of the small proline-rich 2 family of cornified cell envelope proteins. J Biol Chem 273(36):23297–23303CrossRefPubMedGoogle Scholar
  45. Tatsukawa H, Abe N, Ohashi S, Hitomi K (2015) Distribution of transglutaminase family members in mouse whole body sections. Biochem Biophys Res Commun 467(4):1046–1051. doi:10.1016/j.bbrc.2015.10.001 CrossRefPubMedGoogle Scholar
  46. Telci D, Griffin M (2006) Tissue transglutaminase (TG2)—a wound response enzyme. Front Biosci 11:867–882CrossRefPubMedGoogle Scholar
  47. Terrinoni A, Serra V, Codispoti A, Talamonti E, Bui L, Palombo R, Sette M, Campione E, Didona B, Annicchiarico-Petruzzelli M, Zambruno G, Melino G, Candi E (2012) Novel transglutaminase 1 mutations in patients affected by lamellar ichthyosis. Cell Death Dis 3:e416. doi:10.1038/cddis.2012.152 CrossRefPubMedPubMedCentralGoogle Scholar
  48. Verderio E, Nicholas B, Gross S, Griffin M (1998) Regulated expression of tissue transglutaminase in Swiss 3T3 fibroblasts: effects on the processing of fibronectin, cell attachment, and cell death. Exp Cell Res 239(1):119–138. doi:10.1006/excr.1997.3874 CrossRefPubMedGoogle Scholar
  49. Verderio EA, Johnson T, Griffin M (2004) Tissue transglutaminase in normal and abnormal wound healing: review article. Amino Acids 26(4):387–404. doi:10.1007/s00726-004-0094-4 CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Wien 2016

Authors and Affiliations

  • Consuelo Pitolli
    • 1
  • Valentina Pietroni
    • 1
  • Lyuben Marekov
    • 2
  • Alessandro Terrinoni
    • 3
  • Kiyofumi Yamanishi
    • 4
  • Cinzia Mazzanti
    • 3
  • Gerry Melino
    • 1
  • Eleonora Candi
    • 1
    • 3
  1. 1.Department of Experimental Medicine and SurgeryUniversity of Rome “Tor Vergata”RomeItaly
  2. 2.National Institutes of HealthBethesdaUSA
  3. 3.Biochemistry LaboratoryIDI-IRCCSRomeItaly
  4. 4.Department of DermatologyHyogo College of MedicineNishinomiyaJapan

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