Analysis of the defence phosphoproteome of Arabidopsis thaliana using differential mass tagging
Analysis of the defence phosphoproteome of Arabidopsis thaliana using differential mass tagging
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DOI：
10.1002/pmic.200500172
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发表时间：
2006-07-01
期刊：
影响因子：
3.4
通讯作者：
Grant, Murray
中科院分区：
文献类型：
--
作者：
Jones, Alexandra M. E.;Bennett, Mark H.;Grant, Murray
尽管蛋白质组技术最近取得了进展，但蛋白质组的定量分析仍然是一项具有挑战性的任务。蛋白质的磷酸化是信号转导途径的中心，在植物抵御病原体的防御中发挥着重要作用，尽管激酶的直接靶标仍然难以确定。确定在防御反应过程中磷蛋白质组的变化是分子植物病理学的一个主要目标。在首次描述了应用于植物病原菌相互作用的新的质量标签策略(iTRAQ(TM)应用生物系统)中，我们描述了拟南芥在对丁香假单胞菌PV的防御反应中磷蛋白质组的早期变化。番茄DC3000。我们鉴定了五种蛋白质，这些蛋白质在对照和三种不同的细菌挑战之间显示了可复制的差异，从而识别了潜在的磷酸化蛋白质，作为植物基础防御反应的一部分。这五种蛋白质中的四种，即脱水蛋白、可能的p23辅伴蛋白、热休克蛋白81和叶绿体相关蛋白(PAP)/纤维蛋白，已知是被磷酸化的或具有潜在的磷酸化位点。另一种蛋白质，Rubisco的大亚基，在经历过敏反应和基础防御反应的组织中显示出显著的差异。我们记录了与这种方法相关的重复性、实用性和问题。
Despite recent advances in proteomic technologies, quantitative analysis of the proteome remains a challenging task. Phosphorylation of proteins is central to signal transduction pathways and plays an important role in plant defence against pathogens, although the immediate targets of kinases remain elusive. Determining changes in the phosphoproteome during the defence response is a major goal in molecular plant pathology. In this first description of the novel mass tagging strategy (iTRAQ (TM) Applied Biosystems) applied to plant pathogen interactions, we describe early changes to the phosphoproteome of Arabidopsis thaliana during the defence response to Pseudomonas syringae pv. tomato DC3000. We identified five proteins which showed reproducible differences between a control and three different bacterial challenges, thus identifying proteins potentially phosphorylated as part of a plant basal defence response. Four of the five proteins a dehydrin, a putative p23 co-chaperone, heat shock protein 81 and a plastid-associated protein (PAP)/fibrillin, are known to be phosphorylated or have potential phosphorylation sites. One further protein, the large subunit of Rubisco, showed a significant difference between tissue undergoing the hypersensitive response and a basal defence response. We document the reproducibility, utility and problems associated with this approach.