Tanaka N, Konishi H, Khan M M K, Komatsu S
Department of Molecular Genetics, National Institute of Agrobiological Sciences, 305-8602 Tsukuba, Japan.
Mol Genet Genomics. 2004 Jan;270(6):485-96. doi: 10.1007/s00438-003-0929-9. Epub 2003 Nov 21.
Protein databases constructed using high-resolution two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) were used to explore the proteome expressed in various rice tissues. Proteins from leaf sheath, root, and cultured suspension cells were systematically analyzed using 2D-PAGE, mass spectrometry and Edman sequencing, followed by database searching. In all, 79 of the 431 spots detected by 2D-PAGE in the leaf sheath, 73 of the 508 spots in the root and 140 of the 962 spots in the cultured suspension cells could be identified. Protein lists were constructed for each tissue and used to investigate the effects of gibberellin (GA) treatment. In the leaf sheath, root and cultured suspension cells, 8, 21, and 14 of the identified proteins, respectively, were regulated by GA. These proteins included polypeptides involved in general metabolism, energy production, transcriptional regulation and signal transduction in the leaf sheath; in metabolism and defense in the root; and in metabolism, energy production, cell growth, defense and signal transduction in the cultured suspension cells. These results indicate that the proteome databases assembled in these studies will be useful for the rapid assessment of changes in protein content in specific tissues, and that proteins regulated by GA may play a significant role in tissue growth.
利用高分辨率二维聚丙烯酰胺凝胶电泳(2D-PAGE)构建的蛋白质数据库,用于探索在各种水稻组织中表达的蛋白质组。使用2D-PAGE、质谱分析和埃德曼测序对来自叶鞘、根和培养悬浮细胞的蛋白质进行系统分析,随后进行数据库搜索。总共,在叶鞘中通过2D-PAGE检测到的431个斑点中的79个、根中508个斑点中的73个以及培养悬浮细胞中962个斑点中的140个可以被鉴定出来。为每个组织构建了蛋白质列表,并用于研究赤霉素(GA)处理的效果。在叶鞘、根和培养悬浮细胞中,分别有8个、21个和14个已鉴定的蛋白质受GA调节。这些蛋白质包括参与叶鞘中一般代谢、能量产生、转录调控和信号转导的多肽;根中的代谢和防御;以及培养悬浮细胞中的代谢、能量产生、细胞生长、防御和信号转导。这些结果表明,在这些研究中组装的蛋白质组数据库将有助于快速评估特定组织中蛋白质含量的变化,并且受GA调节的蛋白质可能在组织生长中发挥重要作用。