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Fturtional analyses of bio-cross-Iinking enzyme transglutaminase

生物交联酶转谷氨酰胺酶的功能分析

基本信息

批准号：
18580092
负责人：
IKURA Koji
金额：
$ 2.51万
依托单位：
Kyoto Institute of Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

This research project was carried out to analyze the physiological functions of protein cross-linking enzyme transglutaminase (TG). 1. In rat liver extract, arginase-I (AI) and fructose 1, 6-bisphosphatase (FBPase) were identified as new protein substrate candidates of TG. Activities of both enzymes decreased through the amine incorporation into these enzyme proteins by TG, suggesting the possibility of an involvement of TG in the regulation of glucose and arginine metabolisms. 2. Human brain proteins that interact with human brain TG(hbTG) were searched by use of yeast two-hybrid method. Low molecular weight G proteins Rin and Rit were identified as interacting protein candidates, but their interactions with hbTG were not confirmed by the pull-down assay. 3. Functional analysis of Drosophila TG (dTG) was performed. dTG gene seemed to be transcribed throughout all developmental stages. Knockdown of dTG in the whole body resulted in lethality at the late stage of pupa, indicating that dTG is essential for Drosophila development Over-expression of dTG at eye imaginal discs by GAL4-UAS targeted expression system caused rough eye phenotype. The induced rough eye phenotype is probably attributable to apoptosis caused through the activation of JNK signaling. Over-expression and knockdown of dTG at wing imaginal discs caused extra wing veins and blistered wings, respectively. These resulats indicated that the expression level of dTG in wing imaginal discs may affect the development of wing veins.

本研究旨在分析蛋白质交联酶转氨酶（TG）的生理功能。1.在大鼠肝提取物中，鉴定出胰蛋白酶I（AI）和果糖1，6-二磷酸酶（FBPase）为TG的新的候选蛋白底物。这两种酶的活性下降，通过胺掺入到这些酶蛋白的TG，这表明可能参与的TG在葡萄糖和精氨酸代谢的调节。2.利用酵母双杂交技术寻找与人脑TG相互作用的人脑蛋白。低分子量G蛋白Rin和Rit被鉴定为相互作用蛋白候选物，但它们与hbTG的相互作用未通过下拉测定证实。3.对果蝇TG（dTG）进行了功能分析。dTG基因在整个发育阶段都有转录。dTG基因在果蝇蛹期晚期被敲低导致果蝇死亡，表明dTG是果蝇发育所必需的。利用GAL 4-UAS靶向表达系统在果蝇眼胚盘过量表达dTG，导致果蝇出现粗糙眼表型。诱导的粗糙眼表型可能归因于通过激活JNK信号传导引起的细胞凋亡。dTG在翅盘的过表达和敲低分别导致翅静脉增多和翅红。这些结果表明，dTG在翅盘中的表达水平可能影响翅脉的发育。