In Situ Detection of Active Transglutaminases for Keratinocyte Type (TGase 1) and Tissue Type (TGase 2) Using Fluorescence-Labeled Highly Reactive Substrate Peptides
In Situ Detection of Active Transglutaminases for Keratinocyte Type (TGase 1) and Tissue Type (TGase 2) Using Fluorescence-Labeled Highly Reactive Substrate Peptides
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DOI：
10.1369/jhc.2010.957225
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发表时间：
2011-02-01
影响因子：
3.2
通讯作者：
Hitomi, Kiyotaka
中科院分区：
文献类型：
--
作者：
Itoh, Miho;Kawamoto, Tadafumi;Hitomi, Kiyotaka
转氨酶是一种钙依赖性酶，其通过谷氨酰胺和赖氨酸残基之间的交联或伯胺与特定多肽结合的谷氨酰胺残基的连接来后修饰蛋白质。八种同工酶在哺乳动物的各种生物学过程中起着重要的作用。作者最近鉴定了12个氨基酸的优选底物肽序列，其具有高度反应性并以同工酶特异性方式起作用。在这项研究中，一个快速，同工酶特异性，和敏感的检测活性角质形成细胞类型（TGase 1）和组织类型（TGase 2）是成功的使用荧光标记的肽。该程序涉及使用小鼠的全身切片来广泛分析两种酶的组织分布，这些酶显示出明显不同的模式。在上皮组织如舌、发育中的牙齿、前胃和皮肤表皮中观察到强活性的TGase 1。如预测的，在各种类型的组织中观察到显著活性的TGase 2，并且在肠粘膜、肌膜和肝脏的整个静脉中以特别高的水平观察到。本手稿包含在线补充材料，请访问http://www.jhc.org在线查看这些材料。（J Histochem Cytochem 59：180-187，2011）
Transglutaminase is a calcium-dependent enzyme that posttranslationally modifies proteins by cross-linking between glutamine and lysine residues or attachment of a primary amine to specific polypeptide-bound glutamine residues. Eight isozymes play essential roles in various mammalian biological processes. The authors have recently identified 12-amino acid preferred substrate peptide sequences that are highly reactive and act in an isozyme-specific manner. In this study, a rapid, isozyme-specific, and sensitive detection of active keratinocyte type (TGase 1) and tissue type (TGase 2) was successful using fluorescence-labeled peptides. This procedure involved using whole-body sections of a mouse to extensively analyze the tissue distribution of both enzymes that revealed clearly distinct patterns. Strong active TGase 1 was observed in epithelial tissues such as tongue, developing teeth, forestomach, and skin epidermis. Significantly active TGase 2 was observed in various types of tissues as predicted and at particularly higher levels in the intestinal mucosa, muscle membrane, and whole veins in the liver. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials. (J Histochem Cytochem 59:180-187, 2011)