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プロテオミクスの手法を用いた新規プロリン異性化酵素の機能解析

使用蛋白质组学技术对新型脯氨酸异构酶进行功能分析

基本信息

批准号：
03J06057
负责人：
藤山沙理
金额：
$ 1.02万
依托单位：
Tokyo University of Agriculture and Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for JSPS Fellows
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

项目摘要

本研究では、機能未知タンパク質であるヒトプロリン異性化酵素(PPIase)Par14の機能解析を行ってきた。Par14は、細胞内において有糸分裂、転写、癌化等様々な現象で重要な役割を果たしているヒトPin1と高い相同性を持つことから、Pin1同様細胞内で重要な役割を担うことが予想されているが、その機能は全くわかっていない。そこでまず始めに、機能プロテオミクス解析の手法を用いた機能解析を行い、Par14複合体の構成成分としてリボソームタンパク質、トランス作用因子およびpre-rRNAを同定した。これらの因子は、細胞内においてリボソーム生合成経路の中間体として生成するリボソーム前駆体を構成することから、Par14と相互作用する複合体はリボソーム前駆体であると結論付けた(Fujiyama et al.,The Journal of Biological Chemistry, vol 277,p23773-23780,2002)。さらに、Par14が細胞内でリボソーム生合成経路において機能していることを示すため、siRNAの導入によりPar14をノックダウンした細胞のリボソームプロファイルを調べた。その結果、Par14タンパク質の減少に伴い、60S,80Sサブユニットの減少が認められた。このことから、Par14は60Sサブユニットの合成を正に制御している可能性が示唆された。以上のように本研究では、プロテオミクスの手法およびsiRNAを用いた解析を行い、Par14が細胞内においてリボソーム生合成を正に制御する可能性を示した。近年、癌化やその他の疾病とリボソーム合成との関連性が示唆されているが、ほ乳類のリボソーム生合成経路の詳細は、その複雑さからほとんど理解されていない。今後Par14の解析をさらに進めることにより、リボソーム合成経路と疾病との関連性を理解するための多くの知見が得られると期待できる。

In this study, we don't know whether or not we can use the PPIase Par14 machine to analyze the bank data. There are many diseases such as division, writing, carcinogenesis and so on in Par14 and cytosol. They are similar to each other in that they are the same as those in Pin1. They are the same as those in the cytosol. They are the same as those in the cytosol. The components of the Par14 complex were analyzed by the computer, and the effect factors were the same as those of the pre-rRNA assay. In the synthetic pathway, the growth factor, the intracellular factor, the synthesis pathway, the biosynthesis pathway, the synthesis pathway, the synthesis pathway, the growth factor, the synthesis pathway, the synthesis pathway, the The pathways of synthesis in the cells of Par14 and Par14 were detected by the computer. The machine was able to display the signals of the Par14 cells and the cells of the cells in the cell. The results were as follows: Par14, 80s, 60s, 80s, 80s, 60s, 80s, 80s, 60s, 80s, 80s, 60s, 80s, 80s, 80s, 60s, 80s, 60s, 80s, 80s, 60s, 80s, 60s, 80s, 80s, 60s, 80s, 60s, 80s, 60s, 80s, 80s, 60s, 80s, 60s, 80s, 80s, 80s, 60s, 80s, 80s, 60s, 80s, 60s, 80s, 60s, 80s, 80s, 80s, 60s, 80s, 80s, 80s, 80s, 60s, 80s, 80s, 80s, 60s, 80s, 80s, 80s, 60s, 80s, 80s, The combination of Par14 60s and 60s is in the process of controlling the possibility of instigation. In this study, siRNA was used in this study, and Par14 was used in this study to determine the possibility of synthesis. In recent years, cancer and other diseases have caused significant changes in the synthesis of breast cancer, breast cancer and breast cancer. In the future, Par14 will analyze the relationship between diseases and diseases in the future.