Full dynamic range proteome analysis of S. cerevisiae by targeted proteomics.

Full dynamic range proteome analysis of S. cerevisiae by targeted proteomics.
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DOI:
10.1016/j.cell.2009.05.051
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发表时间:
2009-08-21
期刊:
影响因子:
64.5
通讯作者:
Aebersold R
Aebersold R
中科院分区:
生物学1区
文献类型:
--
作者:
Picotti P;Bodenmiller B;Mueller LN;Domon B;Aebersold R

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系统生物学的兴起意味着对高灵敏度技术的需求不断增长,以快速、一致地检测和定量多个样品中的目标蛋白质组。这只能通过经典质谱或基于亲和力的方法部分实现。我们应用基于选择反应监测 (SRM) 的靶向蛋白质组学方法来检测和定量总酿酒酵母消化物中表达浓度低于 50 个拷贝/细胞的蛋白质。检测范围可以扩展到个位数拷贝/细胞以及经典方法无法检测到的蛋白质。我们通过对跨越一系列代谢阶段的酿酒酵母生长时间过程中的整个丰度范围的蛋白质网络进行一致和快速的测量来说明该技术的威力。因此,我们证明了基于 SRM 的蛋白质组学的潜力,可以提供以高通量和定量精度测量酵母中任何感兴趣的蛋白质组的分析方法。
The rise of systems biology implied a growing demand for highly sensitive techniques for the fast and consistent detection and quantification of target sets of proteins across multiple samples. This is only partly achieved by classical mass spectrometry or affinity-based methods. We applied a targeted proteomics approach based on selected reaction monitoring (SRM) to detect and quantify proteins expressed to a concentration below 50 copies/cell in total S. cerevisiae digests. The detection range can be extended to single-digit copies/cell and to proteins which were undetected by classical methods. We illustrate the power of the technique by the consistent and fast measurement of a network of proteins spanning the entire abundance range over a growth time-course of S. cerevisiae transiting through a series of metabolic phases. We therefore demonstrate the potential of SRM-based proteomics to provide assays for the measurement of any set of proteins of interest in yeast at high-throughput and quantitative accuracy.
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