Response to biotic and oxidative stress in Arabidopsis thaliana: Analysis of variably phosphorylated proteins
Response to biotic and oxidative stress in Arabidopsis thaliana: Analysis of variably phosphorylated proteins
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DOI：
10.1016/j.jprot.2011.05.016
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发表时间：
2011-09-06
影响因子：
3.3
通讯作者：
Marra, Mauro
中科院分区：
文献类型：
--
作者：
Huang, Chao;Verrillo, Francesca;Marra, Mauro
蛋白质磷酸化在许多细胞事件的调控中起着关键作用，越来越多的证据表明这种翻译后修饰参与了植物对各种非生物和生物胁迫的应答。由于磷酸化蛋白可能以低丰度存在，因此通常需要富集方法对其进行分析。在这里，我们描述了磷蛋白的定量变化，目前在拟南芥叶片后的挑战与激发子或治疗模拟生物应力，刺激基础的电阻反应，或氧化应激。通过金属氧化物亲和层析富集来自诱导和对照植物的磷蛋白，并通过2D电泳解析。所得蛋白质组图谱的比较突出了显示由激发子处理诱导的定量变化的磷蛋白;这些组分通过MALDI-TOF肽质量指纹图谱和/或nanoLC-ESI-LIT-MS/MS实验鉴定。总共有97个差异点，代表75个独特的候选磷蛋白，进行了表征。它们代表不同的蛋白质功能组，例如能量和碳代谢，对氧化和非生物胁迫的响应，防御，蛋白质合成，RNA加工和细胞信号传导。在拟南芥磷酸化蛋白质组数据库中得到了肯定的证实。每种磷蛋白的作用在这里讨论与植物防御机制。我们的研究结果表明，对不同治疗的反应部分重叠，以及通过施加压力与关键细胞功能进行通信。(C)2011 Elsevier B. V.保留所有权利。
Protein phosphorylation plays a pivotal role in the regulation of many cellular events; increasing evidences indicate that this post-translational modification is involved in plant response to various abiotic and biotic stresses. Since phosphorylated proteins may be present at low abundance, enrichment methods are generally required for their analysis. We here describe the quantitative changes of phosphoproteins present in Arabidopsis thaliana leaves after challenging with elicitors or treatments mimicking biotic stresses, which stimulate basal resistance responses, or oxidative stress. Phosphoproteins from elicited and control plants were enriched by means of metal oxide affinity chromatography and resolved by 2D electrophoresis. A comparison of the resulting proteomic maps highlighted phosphoproteins showing quantitative variations induced by elicitor treatment; these components were identified by MALDI-TOF peptide mass fingerprinting and/or nanoLC-ESI-LIT-MS/MS experiments. In total, 97 differential spots, representing 75 unique candidate phosphoproteins, were characterized. They are representative of different protein functional groups, such as energy and carbon metabolism, response to oxidative and abiotic stresses, defense, protein synthesis, RNA processing and cell signaling. Ascertained protein phosphorylation found a positive confirmation in available Arabidopsis phosphoproteome database. The role of each identified phosphoprotein is here discussed in relation to plant defense mechanisms. Our results suggest a partial overlapping of the responses to different treatments, as well as a communication with key cellular functions by imposed stresses. (C) 2011 Elsevier B.V. All rights reserved.