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

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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.

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Abbreviations

TGs:

Transglutaminases

ECM:

Extracellular matrix

KO:

Knock-out mouse

DKO:

Double knock-out mouse

CE:

Cornified cell envelope

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Acknowledgements

This work was mainly supported by “Ricerca Corrente RC-1” Italian Ministry of Health to AT, and partially supported by the Medical Research Council (MRC), “Fondazione Roma” NCD Grant to GM and EC.

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    Authors and Affiliations

    1. Department of Experimental Medicine and Surgery, University of Rome “Tor Vergata”, Rome, Italy

      Consuelo Pitolli, Valentina Pietroni, Gerry Melino & Eleonora Candi

    2. National Institutes of Health, Bethesda, MD, USA

      Lyuben Marekov

    3. Biochemistry Laboratory, IDI-IRCCS, Rome, Italy

      Alessandro Terrinoni, Cinzia Mazzanti & Eleonora Candi

    4. Department of Dermatology, Hyogo College of Medicine, Nishinomiya, Hyogo, 663-8501, Japan

      Kiyofumi Yamanishi

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    1. Consuelo Pitolli
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    2. Valentina Pietroni
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    4. Alessandro Terrinoni
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    Correspondence to Eleonora Candi.

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    Handling Editors: S. Beninati, M. Piacentini, C. M. Bergamini.

    C. Pitolli and V. Pietroni contributed equally.

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    Pitolli, C., Pietroni, V., Marekov, L. et al. Characterization of TG2 and TG1–TG2 double knock-out mouse epidermis. Amino Acids 49, 635–642 (2017). https://doi.org/10.1007/s00726-016-2356-3

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