Picotti Paola, Bodenmiller Bernd, Mueller Lukas N, Domon Bruno, Aebersold Ruedi
Institute of Molecular Systems Biology, ETH Zurich, Zurich CH 8093, Switzerland.
Cell. 2009 Aug 21;138(4):795-806. doi: 10.1016/j.cell.2009.05.051. Epub 2009 Aug 6.
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 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.
系统生物学的兴起意味着对高灵敏度技术的需求日益增长，这些技术能够在多个样本中快速、一致地检测和定量目标蛋白质组。经典的质谱法或基于亲和力的方法仅部分满足了这一需求。我们应用了一种基于选择反应监测（SRM）的靶向蛋白质组学方法，以检测和定量在酿酒酵母总消化物中表达浓度低于50拷贝/细胞的蛋白质。检测范围可以扩展到个位数拷贝/细胞以及经典方法未检测到的蛋白质。我们通过在酿酒酵母经历一系列代谢阶段的生长时间过程中，对跨越整个丰度范围的蛋白质网络进行一致且快速的测量，展示了该技术的强大功能。因此，我们证明了基于SRM的蛋白质组学具有在高通量和定量准确性方面为测量酵母中任何感兴趣的蛋白质组提供检测方法的潜力。
