A draft map of the mouse pluripotent stem cell spatial proteome.
A draft map of the mouse pluripotent stem cell spatial proteome.
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DOI：
10.1038/ncomms9992
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发表时间：
2016-01-12
影响因子：
16.6
通讯作者：
Lilley KS
中科院分区：
文献类型：
--
作者：
Christoforou A;Mulvey CM;Breckels LM;Geladaki A;Hurrell T;Hayward PC;Naake T;Gatto L;Viner R;Martinez Arias A;Lilley KS
了解蛋白质的亚细胞分布对于理解细胞机制至关重要。在单个实验中捕获亚细胞蛋白质组已被证明具有挑战性，研究集中在特定的隔室或以低分辨率和/或准确度将蛋白质分配到亚细胞小生境。在这里，我们介绍hyperLOPIT，一种将广泛的分馏，定量高分辨率精确质谱与多变量数据分析相结合的方法。我们将hyperLOPIT应用于亚细胞蛋白质组尚未被广泛研究的多能干细胞群体。我们以前所未有的空间分辨率提供5，000多种蛋白质的定位数据，以揭示细胞器的组织，亚细胞器区室，蛋白质复合物，功能网络和蛋白质的稳态动力学以及意想不到的亚细胞位置。该方法为表征转录后和翻译后修饰对蛋白质位置的影响以及涉及蛋白质组水平位置变化对细胞扰动的研究铺平了道路。一个互动的开源资源，使这些数据的探索。 蛋白质在细胞内的空间位置是蛋白质功能的关键要素。在这里，作者描述了hyperLOPIT-一种蛋白质组学工作流程，允许以高分辨率同时将数千种蛋白质分配到亚细胞小生境-并将其应用于小鼠多能干细胞。
Knowledge of the subcellular distribution of proteins is vital for understanding cellular mechanisms. Capturing the subcellular proteome in a single experiment has proven challenging, with studies focusing on specific compartments or assigning proteins to subcellular niches with low resolution and/or accuracy. Here we introduce hyperLOPIT, a method that couples extensive fractionation, quantitative high-resolution accurate mass spectrometry with multivariate data analysis. We apply hyperLOPIT to a pluripotent stem cell population whose subcellular proteome has not been extensively studied. We provide localization data on over 5,000 proteins with unprecedented spatial resolution to reveal the organization of organelles, sub-organellar compartments, protein complexes, functional networks and steady-state dynamics of proteins and unexpected subcellular locations. The method paves the way for characterizing the impact of post-transcriptional and post-translational modification on protein location and studies involving proteome-level locational changes on cellular perturbation. An interactive open-source resource is presented that enables exploration of these data. The spatial location of proteins within a cell is a key element of protein function. Here the authors describe hyperLOPIT—a proteomics workflow that allows the simultaneous assignment of thousands of proteins to subcellular niches with high resolution—and apply it to mouse pluripotent stem cells.