Proteomics of the rice cell: systematic identification of the protein populations in subcellular compartments
Proteomics of the rice cell: systematic identification of the protein populations in subcellular compartments
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DOI：
10.1007/s00438-004-1002-z
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发表时间：
2004-06-01
影响因子：
3.1
通讯作者：
Komatsu, S
中科院分区：
文献类型：
--
作者：
Tanaka, N;Fujita, M;Komatsu, S
尽管最近在水稻全基因组测序方面取得了进展，但该物种的蛋白质组仍然知之甚少。为了扩大我们对水稻蛋白质组的了解，从水稻幼苗和培养的悬浮细胞中纯化了包括质膜(PM)、液泡膜(Vm)、高尔基膜(GM)、线粒体(MT)和叶绿体(CP)在内的亚细胞亚组。然后使用二维(2D)电泳法、质谱仪和Edman测序法对每个隔室的蛋白质进行系统分析，然后进行数据库搜索。总体而言，464个斑点中的58个、VM的141个斑点中的43个、GM的361个斑点中的46个、MT的672个斑点中的146个以及CP的252个斑点中的89个可以通过该程序识别。研究发现，这些蛋白质参与了不同的过程，如MT的呼吸作用和柠檬酸循环，CP的光合作用和ATP合成，以及膜上的抗逆防御和信号系统。Edman降解显示，60%-98%的N-末端序列被阻断，并且在不同亚细胞隔室的蛋白质组中，封闭和未阻断的蛋白质的比例不同。水稻亚细胞室蛋白质组的数据对于解决功能基因组学中的问题以及对植物功能的全基因组探索将是有价值的。
Despite recent progress in sequencing the complete genome of rice (Oryza saliva), the proteome of this species remains poorly understood. To extend our knowledge of the rice proteome, the subcellular compartments, which include plasma membranes (PM), vacuolar membranes (VM), Golgi membranes (GM), mitochondria (MT), and chloroplasts (CP), were purified from rice seedlings and cultured suspension cells. The proteins of each of these compartments were then systematically analyzed using two-dimensional (2D) electrophoresis, mass spectrometry, and Edman sequencing, followed by database searching. In all, 58 of the 464 spots detected by 2D electrophoresis in PM, 43 of the 141 spots in VM, 46 of the 361 spots in GM, 146 in the 672 spots in MT, and 89 of the 252 spots in CP could be identified by this procedure. The characterized proteins were found to be involved in various processes, such as respiration and the citric acid cycle in MT; photosynthesis and ATP synthesis in CP; and antiftingal defense and signal systems in the membranes. Edman degradation revealed that 60-98% of N-terminal sequences were blocked, and the ratios of blocked to unblocked proteins in the proteomes of the various subcellular compartments differed. The data on the proteomes of subcellular compartments in rice will be valuable for resolving questions in functional genomics as well as for genome-wide exploration of plant function.