Rice proteome analysis as a post-genome project research

为了有效地分析水稻蛋白质组，本研究开发了多种分析技术。这些技术包括凝胶内蛋白质消化技术、肽图谱技术、用于鉴定二维电泳分离的蛋白质的质谱技术、用于鉴定SDS凝胶电泳分离的蛋白质的散弹枪分析技术、磷酸化蛋白质检测技术、高分子量蛋白质电泳技术以及使用新型蛋白质芯片进行蛋白质-蛋白质相互作用分析技术。利用所开发的技术，对水稻种子胚蛋白质组进行了全面分析。在这个分析中，我们发现了许多没有已知功能的蛋白质。为了获得这些蛋白的功能信息，我们对它们进行了表达谱分析，并鉴定了几个与发芽有关的表达蛋白。另一方面，我们分析了一个大的蛋白质复合物26S蛋白酶体，以确定蛋白质翻译后修饰的作用，为蛋白质组学分析获得基础信息。在蛋白质组数据库中编制了双向电泳图谱和单个胚胎蛋白的特征。

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.