Studies on differentiation-dependent expression and activity of distinct transglutaminases by specific substrate peptides using three dimensional reconstituted epidermis.
Studies on differentiation-dependent expression and activity of distinct transglutaminases by specific substrate peptides using three dimensional reconstituted epidermis.
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使用三维重建表皮研究特定底物肽对不同转谷氨酰胺酶的分化依赖性表达和活性。
DOI：
10.1111/febs.14832
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发表时间：
2019
期刊：
影响因子：
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通讯作者：
Ｔａｎａｂｅ Ｙ．， Ｙａｍａｎｅ Ｍ， Ｋａｔｏ Ｍ．， Ｔｅｓｈｉｍａ Ｈ．， Ｋｕｒｉｂａｙａｓｈｉ Ｍ．， Ｔａｔｓｕｋａｗａ Ｈ．， Ｔａｋａｍａ Ｈ．， Ａｋｉｙａｍａ Ｍ．， Ｈｉｔｏｍｉ Ｋ．
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文献类型：
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作者：
Masataka Sugiyama;Yasuo Takehara;Ryota Horiguchi;Takashi Mizuno;Ryota Hyodo;Takasuke Ushio;Tetsuya Wakayama;Atsushi Nozaki;Hiroyuki Kabasawa;Marcus Alley;Satoshi Goshima;Shinji Naganawa;Ｔａｎａｂｅ Ｙ．， Ｙａｍａｎｅ Ｍ， Ｋａｔｏ Ｍ．， Ｔｅｓｈｉｍａ Ｈ．， Ｋｕｒｉｂａｙａｓｈｉ Ｍ．， Ｔａｔｓｕｋａｗａ Ｈ．， Ｔａｋａｍａ Ｈ．， Ａｋｉｙａｍａ Ｍ．， Ｈｉｔｏｍｉ Ｋ．
在皮肤形成过程中，特别是在角质形成细胞的分化过程中，独特的翻译后修饰在形成称为角化包膜（CE）的蛋白质超分子中发挥作用，这是屏障功能所必需的。转谷氨酰胺酶（tgg）是参与各种角化细胞结构蛋白交联以完成CE形成的必需酶。TG家族由8个同工酶组成，其中两个成员TG1和TG3主要位于表皮。在体外三维（3D）培养系统中，表皮的重建使最终分化的角质形成细胞得以凝聚。在这项研究中，我们使用同工酶特异性底物肽来检测TG活性，研究了在该培养体系中分化过程中每种同工酶的表达和激活模式。在分化细胞中，TG1和TG3的蛋白水平、酶活性以及定位表现出不同的模式。siRNA特异性敲除这些酶显示出较少的玉米化，这表明每种TG都有助于表皮的形成。总之，我们证明了3D系统通过特定底物肽研究不同tg的分化依赖性表达和活性的效率。EnzymeTransglutaminase EC2.3.2.13。
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 (TGs) 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, TG1 and TG3, located mainly in the epidermis. In anin vitrothree‐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 TG1 and TG3 exhibited distinct patterns. Specific knockdown of these enzymes by siRNA 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 TGs by specific substrate peptides.EnzymeTransglutaminase, EC2.3.2.13.