Mapping the Arabidopsis organelle proteome
Mapping the Arabidopsis organelle proteome
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DOI：
10.1073/pnas.0506958103
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发表时间：
2006-04-25
影响因子：
11.1
通讯作者：
Lilley, KS
中科院分区：
文献类型：
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作者：
Dunkley, TPJ;Hester, S;Lilley, KS
在分泌途径的研究中，一个具有挑战性的任务是鉴定和定位新的蛋白质，以增加我们对不同细胞器功能的理解。以前对细胞膜系统的蛋白质组学研究一直受到蛋白质污染的阻碍，这使得无法将蛋白质分配给细胞器。在这里，我们使用同位素标记技术来定位细胞器蛋白质，结合同位素标记进行相对和绝对定量，并使用二维液相色谱法同时将蛋白质分配到多个亚细胞区室。利用这种方法，测定了拟南芥689种蛋白的密度梯度分布，使527种蛋白能够同时定位于内质网、高尔基体、空泡膜、质膜或线粒体和质体。这种膜成分的平行分析使蛋白质的稳态分布得以确定。因此，真正的细胞器居民已经与污染蛋白质和通过分泌途径转运的蛋白质区分开来。
A challenging task in the study of the secretory pathway is the identification and localization of new proteins to increase our understanding of the functions of different organelles. Previous proteomic studies of the endomembrane system have been hindered by contaminating proteins, making it impossible to assign proteins to organelles. Here we have used the localization of organelle proteins by the isotope tagging technique in conjunction with isotope tags for relative and absolute quantitation and 2D liquid chromatography for the simultaneous assignment of proteins to multiple subcellular compartments. With this approach, the density gradient distributions of 689 proteins from Arabidopsis thaliana were determined, enabling confident and simultaneous localization of 527 proteins to the endoplasmic reticulum, Golgi apparatus, vacuolar membrane, plasma membrane, or mitochondria and plastids. This parallel analysis of endomembrane components has enabled protein steady-state distributions to be determined. Consequently, genuine organelle residents have been distinguished from contaminating proteins and proteins in transit through the secretory pathway.