Variations in Both TG1 and TG2 Isozyme-specific In Situ Activities and Protein Expressions during Mouse Embryonic Development
Variations in Both TG1 and TG2 Isozyme-specific In Situ Activities and Protein Expressions during Mouse Embryonic Development
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DOI：
10.1369/0022155413501676
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发表时间：
2013-11-01
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;Tatsukawa, Hideki;Hitomi, Kiyotaka
转氨酶（TG）是催化蛋白质之间的交联反应的酶家族。使用荧光标记的高反应性底物肽，我们最近开发了一个系统，以可视化同工酶特异性原位酶活性。在本研究中，我们调查了在原位活动的TG1（皮肤型）和TG2（组织型）使用整个小鼠切片的各个胚胎发育阶段和新生儿。在每种情况下，我们还成功地使用相同的整个小鼠切片的免疫染色检测酶活性后的蛋白质表达。一般来说，酶活性与TG蛋白表达相关。然而，在某些组织中，TG蛋白的表达模式，这是不一致的酶的活动，表明无活性的TG可能产生的自交联或其他修饰。我们的方法使我们能够同时观察发育变化的TG同工酶特异性活动和蛋白质水平在小鼠胚胎和新生儿组织。
Transglutaminase (TG) is a family of enzymes that catalyzes cross-linking reactions among proteins. Using fluorescent-labeled highly reactive substrate peptides, we recently developed a system to visualize isozyme-specific in situ enzymatic activity. In the present study, we investigated the in situ activities of TG1 (skin-type) and TG2 (tissue-type) using whole mouse sections of various embryonic developmental stages and neonates. In each case, we also successfully used immunostaining of identical whole mouse sections for protein expression after detection of enzymatic activities. In general, the enzymatic activity was correlated with TG protein expression. However, in some tissues, TG protein expression patterns, which were inconsistent with the enzymatic activities, suggested that inactive TGs were produced possibly by self cross-linking or other modifications. Our method allowed us to simultaneously observe developmental variations in both TG isozyme-specific activities and protein levels in mouse embryonic and neonate tissues.