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Studies on differentiation‐dependent expression and activity of distinct transglutaminases by specific substrate peptides using three‐dimensional reconstituted epidermis
Author(s) -
Tanabe Yuki,
Yamane Miki,
Kato Manami,
Teshima Hirofumi,
Kuribayashi Miki,
Tatsukawa Hideki,
Takama Hiroyuki,
Akiyama Masashi,
Hitomi Kiyotaka
Publication year - 2019
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/febs.14832
Subject(s) - tissue transglutaminase , epidermis (zoology) , keratinocyte , isozyme , microbiology and biotechnology , enzyme , biology , cellular differentiation , biochemistry , substrate (aquarium) , in vitro , involucrin , gene , anatomy , ecology
During skin formation, particularly during differentiation of keratinocytes, unique post‐translational modifications play a role in forming a proteinaceous supermolecule called the cornified envelope ( CE ), which is necessary for barrier function. Transglutaminases ( TG s) are essential enzymes involved in the cross‐linking of various keratinocyte structural proteins to complete CE formation. The TG family consists of eight isozymes, with two members, TG 1 and TG 3, located mainly in the epidermis. In an in vitro three‐dimensional (3D) culture system, reconstruction of the epidermis allows cornification of the terminally differentiated keratinocytes. In this study, using isozyme‐specific substrate peptides that enable detection of TG activity, we investigated the expression and the activation pattern of each isozyme during differentiation in this culture system. In the differentiating cells, the protein levels, enzymatic activities, as well as localization of TG 1 and TG 3 exhibited distinct patterns. Specific knockdown of these enzymes by si RNA revealed less cornification, suggesting that each TG contributes to the epidermal formation. In conclusion, we demonstrate the efficiency of the 3D system for studying differentiation‐dependent expression and activity of distinct TG s by specific substrate peptides. Enzyme Transglutaminase, EC2.3.2.13 .