通过无标记定量磷酸化蛋白质组学揭示病毒诱导的细胞信号级联反应。

Institute of Medical Virology, University of Zurich, 8057 Zurich, Switzerland.

Life Sciences Zurich Graduate School, ETH and University of Zurich, 8057 Zurich, Switzerland.

STAR Protoc. 2022 Jan 17;3(1):101089. doi: 10.1016/j.xpro.2021.101089. eCollection 2022 Mar 18.

Due to the low stoichiometry and highly transient nature of protein phosphorylation it is challenging to capture the dynamics and complexity of phosphorylation events on a systems level. Here, we present an optimized protocol to measure virus-induced phosphorylation events with high sensitivity using label free quantification-based phosphoproteomics. Specifically, we describe filter assisted protein digestion (FASP), enrichment of phosphopeptides, mass spectrometry, and subsequent bioinformatic analysis. For complete details on the use and execution of this protocol, please refer to Hunziker et al. (2022).

由于蛋白质磷酸化的低化学计量和高度瞬态性质，因此很难在系统水平上捕捉磷酸化事件的动态和复杂性。在这里，我们提出了一种使用基于无标记定量的磷酸化蛋白质组学来测量病毒诱导的磷酸化事件的优化方案，具有高灵敏度。具体来说，我们描述了过滤辅助蛋白消化（FASP）、磷酸肽富集、质谱分析以及随后的生物信息学分析。有关该方案使用和执行的完整详细信息，请参阅 Hunziker 等人。（2022 年）。