Identification of a preferred substrate peptide for transglutaminase 3 and detection of in situ activity in skin and hair follicles
Identification of a preferred substrate peptide for transglutaminase 3 and detection of in situ activity in skin and hair follicles
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转谷氨酰胺酶 3 的优选底物肽的鉴定以及皮肤和毛囊中原位活性的检测
DOI：
10.1111/j.1742-4658.2010.07765.x
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
K. Hitomi
中科院分区：
文献类型：
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作者：
A. Yamane;Mina Fukui;Y. Sugimura;Miho Itoh;M. P. Alea;V. Thomas;S. El Alaoui;M. Akiyama;K. Hitomi
转氨酶（TGases）是催化蛋白质之间交联反应的酶家族。在表皮分化过程中，这些酶反应对于角质化包膜的形成至关重要，角质化包膜由交联的结构蛋白组成。两种主要的转氨酶亚型，表皮型（TGase 3）和角质形成细胞型（TGase 1），共同参与角质形成细胞的分化过程。 有关其底物偏好的信息对于确定这些同工酶的功能作用和阐明它们可能的合作作用非常重要。到目前为止，我们已经鉴定了由TGase同工酶如TGase 1、TGase 2（组织型同工酶）和凝血同工酶因子XIII特异性识别的高反应性肽序列。 在这项研究中，从噬菌体展示肽库中筛选了人TGase 3的几种底物肽序列。 选择TGase 3的优选底物序列，并作为与突变型谷胱甘肽S-转移酶的融合蛋白进行评价。 从这些研究中，鉴定了高反应性和同工酶特异性序列（E51）。此外，发现该序列是肽形式的主要底物，并且适合于检测小鼠表皮中的原位TGase 3活性。 在分化的角质形成细胞和毛囊层中检测到TGase 3酶活性，其模式与TGase 1的模式不同。 我们的发现提供了有关TGase 3特定分布的新信息，并构成了阐明其在表皮中功能作用的有用工具。
Transglutaminases (TGases) are a family of enzymes that catalyze cross‐linking reactions between proteins. During epidermal differentiation, these enzymatic reactions are essential for formation of the cornified envelope, which consists of cross‐linked structural proteins. Two main transglutaminases isoforms, epidermal‐type (TGase 3) and keratinocyte‐type (TGase 1), are cooperatively involved in this process of differentiating keratinocytes. Information regarding their substrate preference is of great importance to determine the functional role of these isozymes and clarify their possible co‐operative action. Thus far, we have identified highly reactive peptide sequences specifically recognized by TGases isozymes such as TGase 1, TGase 2 (tissue‐type isozyme) and the blood coagulation isozyme, Factor XIII. In this study, several substrate peptide sequences for human TGase 3 were screened from a phage‐displayed peptide library. The preferred substrate sequences for TGase 3 were selected and evaluated as fusion proteins with mutated glutathione S‐transferase. From these studies, a highly reactive and isozyme‐specific sequence (E51) was identified. Furthermore, this sequence was found to be a prominent substrate in the peptide form and was suitable for detection of in situ TGase 3 activity in the mouse epidermis. TGase 3 enzymatic activity was detected in the layers of differentiating keratinocytes and hair follicles with patterns distinct from those of TGase 1. Our findings provide new information on the specific distribution of TGase 3 and constitute a useful tool to clarify its functional role in the epidermis.