Abstract
Transglutaminase (TGase) is a calcium ion-dependent enzyme that catalyze the formation of isopeptide cross-links in proteins between the γ -carboxamide groups of glutamine residues and ε -amino groups of lysine residues. Both TGases of keratinocyte-type (TGase 1) and epidermal-type (TGase 3) are involved in the formation of the cornified cell envelop, which serve a vital barrier function for the skin, by cross-linking of a variety of structural proteins in the epidermis. TGase 1 and TGase 3 are 107 kDa and 77kDa protein, respectively. Both enzymes are limitedly proteolysed during epidermal differentiation.
It has been difficult to isolate sufficient quantities of native enzymes from tissues or to obtain recombinant proteins in a bacterial expression system for biochemical studies on its properties of the TGases. Therefore, we circumvented these problems by expressing recombinant full-length cDNA of the TGases in baculovirus-infected insect cells. The expressed proteins were purified to homogeneity by successive chromatography and HPLC.
Calpain, that had been a candidate for activating enzyme, was not effective for activation of TGase 3 zymogen. In the case of TGase 1, the calpain-proteolysed form was highly sensitive to the calcium-ion for TGase activity.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsPreview
Unable to display preview. Download preview PDF.
References
Greenberg, C. S., Birckbichler, P. J., and Rice, R. H. (1991) Transglutaminases: multifunctional cross-linking enzymes that stabilize, FASEB Journal, 5, 3071–3077.
Aeschlimann, D., and Paulsson,M. (1994) Transglutaminase:Protein cross-linking enzymes in tissues and body fluids, Thrombosis and Haemostasis, 71, 402–415.
Rice, R. H., Rong, X., and Chakravarty, R. (1990) Proteolytic release of keratinocyte transglutaminase, Biochem.J., 265, 351–357.
Kim, S. Y., Chung, S. I., and Steinert, P. M. (1995) Highly active soluble processed forms of the transglutamiase 1 enzyme in epidermal keratinocytes, J.Biol. Chem., 270, 18026–18035.
Steinert, P. M., Kim, S. Y., Chung, S. I. and Marekov, L. N. (1996) The transglutaminase 1 enzyme is variably acylated by myristate and palmitate during differentiation in epidermal keratinocytes. J. Biol. Chem., 271, 26242–26250.
Kim, H. C., Lewis, M. S., Gorman, J. J., Park, S. C., Girard, J. E., and Folk, J.E. (1990) Protransglutaminase E from guinea pig skin. Isolation and partial characterization, J.Biol. Chem., 265, 21971–21978.
Kim, I. G., Gorman, J. J., Park, S. C., Chung, S. I., and Steinert, P. M. (1993) The deduced sequence of the novel protransglutaminase E of human, J.Biol.Chem., 268, 12682–12690.
Hitomi, K., Kanehiro, S., Ikura, K. and Maki, M. (1999) Characterization of recombinant mouse epidermal-type transglutaminase (TGase 3):Regulation of its activity by proteolysis and gunanine nucleotides. J. Biochem. (Tokyo), 125, 1048–1054.
Hitomi, K., Yamagiwa Y., Ikura, K., Yamanishi K., and Maki, M. (2000) Characterization of human recombinant transglutaminase 1 purified from baculovirus-infected insect cells. Biosci.Biotechnol. Biochem., 64, 2128–2137.
Yamanishi, K., Liew, F. M. Konishi, K., Yasuno, H. Doi, H., Hirano, J. and Fukushima, S. (1991) Molecular cloning of human epidermal transglutaminase cDNA from keratinocyte.Biochem.Biophys. Res. Commun.,175 906–913.
Yamazaki, M. Ishidoh, K., Suga, Y. and Saido. T.C., Kawashinma S., Suzuki K., Kominami E., and Ogawa H. (1997) Cytoplasmic processing of human profilaggrin by active mucalpain. Biochem. Biophys. Res. Commun.,235 652–656.
Kim, S. Y. and Bae, C. D. (1998) Calpain inhibitors reduce the comified cell envelope formation by inhibiting proteolytic processing of TGase 1. Exp. Mol.MedL, 30, 257–262.
Melino, G., Bernassola, F., Knight, R., Corasaniti, M. T., Nistico, G. and Fina7zi-Agro, A. (1997) S-nitrosylation regulates apoptosis. Nature, 388, 432–433.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2002 Springer Science+Business Media Dordrecht
About this chapter
Cite this chapter
Hitomi, K., Ikura, K., Maki, M. (2002). Characterization of Recombinant Transglutaminases 1 and 3 Expressed in Baculovirus System. In: Shirahata, S., Teruya, K., Katakura, Y. (eds) Animal Cell Technology: Basic & Applied Aspects. Animal Cell Technology: Basic & Applied Aspects, vol 12. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-0728-2_30
Download citation
DOI: https://doi.org/10.1007/978-94-017-0728-2_30
Publisher Name: Springer, Dordrecht
Print ISBN: 978-90-481-5934-5
Online ISBN: 978-94-017-0728-2
eBook Packages: Springer Book Archive