Studies on activating mechanisms oftransglutaminase in epidermis

表皮转谷氨酰胺酶激活机制的研究

• 批准号: 12660074
• 负责人: HITOMI Kiyotaka
• 金额: $ 2.56万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12660074/
• 关键词: Transglutaminase; Epidermis; Proteasea; トランスグルタミナ-ゼ

Transglutaminases catalyze the posttranslational modification of proteins by the formation of isopeptide bond either within or between polypeptide chains. This reaction is calcium ion-dependent found in various tissues and cells from mammals to bacteria. In human, nine isozymes with similar primary structure have been discovered so far. Among them, Tgases 1 and 3 are involved in the formation ofcornified envelop by cross-linking various structural proteins. Both enzymes are synthesized as immature proteins and activated during differentiation. The zymogen form enzymes are limitedly proteolyzed for activation. In this study, using recombinant proteins produced and purified in baculos, the responsible proteases for activation were investigated. In respect to the Tgase 1, calpain could proteolyze zymogen form. The proteolyzed molecule was enhanced at the sensitivity of Ca for transglutaminase activity. Furthermore, we also investigated the tissue distribution of Tgase 3 using RT-POR (reverse transcription polymerase chain reaction). In addition to skin and mouse forestomach, Tgase 3 was expressed also in brain, spleen, and testis.

转氨酶通过在多肽链内或之间形成异肽键来催化蛋白质的翻译后修饰。这种反应依赖于钙离子，存在于从哺乳动物到细菌的各种组织和细胞中。在人类中，迄今已发现9种一级结构相似的同工酶。其中，Tgas 1和Tgas 3通过交联各种结构蛋白参与皮质包膜的形成。这两种酶都是作为未成熟蛋白质合成的，并在分化过程中被激活。酶原形式的酶被有限地蛋白水解以活化。在这项研究中，使用重组蛋白生产和纯化的baculos，负责激活蛋白酶进行了研究。对于TGase 1，钙蛋白酶可水解酶原形式。钙对转谷氨酰胺酶活性的敏感性增强了蛋白水解分子。此外，我们还使用RT-POR（逆转录聚合酶链反应）研究了Tgase 3的组织分布。除皮肤和小鼠前胃外，Tgase 3还在脑、脾和睾丸中表达。

项目成果

Ohkouchi et al.: "Identific ati on and Characterization of Two Penta-EF-Hand Ca(2+)-Bmding Proteins in. Dictyostelium discoideum"Journal of Biochemistry. 130. 207-215 (2001)

Ohkouchi等人：“盘基网柄菌中两种Penta-EF-Hand Ca(2)-Bmding蛋白质的鉴定和表征”生物化学杂志。

Hitomi et al.: "Analysis of Epidermal Transglutaminase(TGese 3) Expression in House Tissues and Cell Lines"The International Jounal of Biochemisty and Cell Biology. 33. 491-498 (2001)

Hitomi 等人：“室内组织和细胞系中表皮转谷氨酰胺酶 (TGese 3) 表达的分析”国际生物化学和细胞生物学杂志。

Kitaura et al.: "Peflin and ALG-2, Members of the Penta-EF-Hand Protein Family, Form a Heterodimer that Dissociates in a Ca2+-Dependent Manner"Journal of Biological Chemistry. 276. 14503-14508 (2001)

Kitaura 等人：“Peflin 和 ALG-2，Penta-EF-Hand 蛋白家族的成员，形成以 Ca2 依赖性方式解离的异二聚体”生物化学杂志。

Hitomi et al: "GTP, an Inhibitor of Transglutaminases, is Hydrolyzed by Tissue-type Transglutaminase (Tgase 2) but not by Epidermal-type Transglutaminase (Tgase 3)"Biosci. Biotechnol. Biochem. 64. 657-659 (2000)

Hitomi 等人：“GTP 是一种转谷氨酰胺酶抑制剂，可被组织型转谷氨酰胺酶 (Tgase 2) 水解，但不会被表皮型转谷氨酰胺酶 (Tgase 3) 水解”Biosci。

Ohkouchi et al.: "Identification and characterization of Two Penta-EF-Ca(2+)-Binding proteins in pictyostelium discoideum"Journal of Biochemistry(Tokyo). 130. 207-215 (2001)

Ohkouchi 等人：“盘基网柄菌中两种 Penta-EF-Ca(2)-结合蛋白的鉴定和表征”生物化学杂志（东京）。