Arabidopsis cell wall proteome defined using multidimensional protein identification technology
Arabidopsis cell wall proteome defined using multidimensional protein identification technology
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DOI：
10.1002/pmic.200500046
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发表时间：
2006-01-01
期刊：
影响因子：
3.4
通讯作者：
Maule, AJ
中科院分区：
文献类型：
--
作者：
Bayer, EM;Bottrill, AR;Maule, AJ
随着拟南芥基因组测序的完成和蛋白质组学技术的最新进展，从高度复杂的混合物中鉴定蛋白质现在成为可能。我们没有使用凝胶电泳和肽质量指纹图谱，而是使用多维蛋白质鉴定技术 (MudPIT) 来分析来自拟南芥细胞悬浮培养物的纯化细胞壁的“紧密结合”蛋白质组。使用生物信息学预测靶向分泌途径的信号肽以及在不存在 ER 滞留信号的情况下，选择 89 种蛋白质作为潜在的细胞外蛋白质。在之前对拟南芥细胞壁的蛋白质组学分析中，仅鉴定出其中的 33%。功能分类显示大部分蛋白质是酶，特别是碳水化合物活性酶、过氧化物酶和蛋白酶。比较所有已发表的拟南芥细胞壁蛋白质组学分析，确定了 268 个编码壁蛋白的非冗余基因。其中 60 个 (22%) 来自我们对紧密结合的壁蛋白的分析。
With the completion of the sequencing of the Arabidopsis genome and the recent advances in proteomic technology, the identification of proteins from highly complex mixtures is now possible. Rather than using gel electrophoresis and peptide mass fingerprinting, we have used multidimensional protein identification technology (MudPIT) to analyse the 'tightly-bound' proteome for purified cell walls from Arabidospis cell suspension cultures. Using bioinformatics for the prediction of signal peptides for targeting to the secretory pathway and for the absence of ER retention signal, 89 proteins were selected as potential extracellular proteins. Only 33% of these were identified in previous proteomic analyses of Arabidopsis cell walls. A functional classification revealed that a large proportion of the proteins were enzymes, notably carbohydrate active enzymes, peroxidases and proteases. Comparison of all the published proteomic analyses for the Arabidopsis cell wall identified 268 non-redundant genes encoding wall proteins. Sixty of these (22%) were derived from our analysis of tightly-bound wall proteins.