Rice proteome analysis as a post-genome project research

水稻蛋白质组分析作为后基因组项目研究

• 批准号: 12460004
• 负责人: HIRANO Hisashi
• 金额: $ 9.34万
• 依托单位: Yokohama City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12460004/
• 关键词: Proteome; Protein; Mass spectrometry; Protein function; Post-tranmslational modification; Protein-protein interaction; Seed embryo; Rice; プロテアソーム; 蛋白質機能; 蛋白質の動態; プロテインチップ; 機能; 発芽; プロテオミクス; 胚芽蛋白質; ペプチドマスフィンガープリンティング; アミノ酸配列分析

To analyze efficiently the rice proteome, we developed various analytical techniques in the present study. These include techniques for protein in-gel digestion, peptide mapping, mass spectrometry for identification of proteins separated by two-dimensional electrophoresis, shot-gun analysis of proteins separated by SDS gel electrophoresis, detection for phosphorylated proteins, electrophoresis for high molecular weight proteins, and protein-protein interaction analysis with a novel protein chip. Using the developed techniques, we analyzed comprehensively the rice seed embryo proteome. In this analysis, we identified many proteins without known functions. To obtain the information of functions of these proteins, we performed expression profiling of them, and we identified several proteins, which expressed in relation to germination. On the other hand, we analyzed a large protein complex, 26S proteasome to determine the roles of protein post-translational modifications to obtain fundamental information for proteome analysis. The two-dimensional electrophoresis pattern and the characteristics of individual embryo protein were compiled in our proteome database.

为了有效地分析水稻蛋白质组，本研究发展了多种分析技术。这些技术包括蛋白质凝胶内消化，肽图谱，质谱鉴定的蛋白质分离的二维电泳，鸟枪分析的蛋白质分离的SDS凝胶电泳，检测磷酸化蛋白质，电泳的高分子量蛋白质，蛋白质-蛋白质相互作用分析与一种新的蛋白质芯片。利用该技术对水稻种胚蛋白质组进行了全面的分析。在这项分析中，我们鉴定了许多功能未知的蛋白质。为了获得这些蛋白质的功能信息，我们对它们进行了表达谱分析，并鉴定了几种与发芽相关的蛋白质。另一方面，我们分析了一个大的蛋白质复合体，26 S蛋白酶体，以确定蛋白质翻译后修饰的作用，为蛋白质组分析提供基础信息。将双向电泳图谱和单个胚胎蛋白质的特征编入我们的蛋白质组数据库。

项目成果

Yoshida, Y.: "E3 ubiquitin-ligase that recognizes sugar chains."Nature. 418. 438-442 (2002)

Yoshida, Y.：“识别糖链的 E3 泛素连接酶。”《自然》。