Hyperplexing: a method for higher-order multiplexed quantitative proteomics provides a map of the dynamic response to rapamycin in yeast.
Hyperplexing: a method for higher-order multiplexed quantitative proteomics provides a map of the dynamic response to rapamycin in yeast.
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DOI：
10.1126/scisignal.2002548
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发表时间：
2012-03-27
影响因子：
7.3
通讯作者：
Gygi SP
中科院分区：
文献类型：
--
作者：
Dephoure N;Gygi SP
大规模定量蛋白质组学可以提供细胞蛋白质丰度的近全局视图。然而，实现合理的蛋白质覆盖率和可靠的定量所需的时间、精力和专业知识限制了这项技术的广泛应用。为了充分利用质谱法来阐明生物系统，需要足够的通量来监测各种条件下的动态变化，并使重复分析能够提供统计功效。我们报告了一种简单的方法来增加定量质谱的多路复用能力，这为细胞信号通路的分析提供了一个平台。使用三重代谢标记和六重同量异序标签，我们同时监测了18个样品中蛋白质丰度的变化，对雷帕霉素刺激的酵母进行了六个时间点的生物学三次重复。该数据集提供了数千种酵母蛋白的时间丰度分布，突出了TOR（雷帕霉素靶点）途径的复杂细胞作用。
Large-scale quantitative proteomics can provide a near-global view of cellular protein abundance. Yet, the time, effort, and expertise required to achieve reasonable protein coverage and reliable quantification have limited the broad application of this technology. To fully leverage mass spectrometry for the elucidation of biological systems requires sufficient throughput to monitor dynamic changes across conditions and to enable replicate analysis to provide statistical power. We report a straightforward approach to increase the multiplexing capacity of quantitative mass spectrometry, which provides a platform for the analysis of cellular signaling pathways. Using triplex metabolic labeling and six-plex isobaric tags, we monitored changes in protein abundance from 18 samples simultaneously, performing biological triplicates of a six-point time course of rapamycin-stimulated yeast. The data set provides temporal abundance profiles for thousands of yeast proteins, highlighting the complex cellular roles of the TOR (target of rapamycin) pathway.