Interactome Profiling of DNA Damage Response (DDR) Mediators with Immunoprecipitation-Mass Spectrometry.
Interactome Profiling of DNA Damage Response (DDR) Mediators with Immunoprecipitation-Mass Spectrometry.
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使用免疫沉淀-质谱法对 DNA 损伤反应 (DDR) 介质进行相互作用组分析。
DOI：
10.1007/978-1-0716-3373-1_12
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Woods,NicholasT
中科院分区：
文献类型：
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作者：
Law,HenryC-H;Noe,Dragana;Woods,NicholasT
免疫沉淀-质谱 (IP-MS) 是一种用于探测生物样品中整体蛋白质-蛋白质相互作用 (PPI) 的多功能工具。这种相互作用协调复杂的生物过程，例如 DNA 损伤反应 (DDR)。 DNA 损伤的诱导会激活信号网络，其中翻译后修饰会导致 PPI，从而促进 DNA 修复和细胞周期协调。 DDR 传感器、传感器和效应器的蛋白质相互作用组分析有可能识别新的 DDR 机制，从而促进我们对与 DDR 缺陷相关的疾病（例如癌症）的理解和治疗。这里描述的协议是常规 PPI 分析程序，可以对受 DNA 损伤刺激的样品执行。所有流程和试剂均经过优化，可实现相互作用组的最大灵敏度和质谱仪的最小污染。
Immunoprecipitation-mass spectrometry (IP-MS) is a versatile tool to probe for global protein–protein interactions (PPIs) in biological samples. Such interactions coordinate complex biological processes, such as the DNA damage response (DDR). Induction of DNA damage activates signaling networks where posttranslational modifications cause PPI that facilitate DNA repair and cell cycle coordination. Protein interactome profiling of DDR sensors, transducers, and effectors has the potential to identify novel DDR mechanisms that could advance our understanding and treatment of diseases associated with DDR defects, such as cancer. The protocol described here is a routine PPI analysis procedure that can be performed on samples stimulated with DNA damage. All processes and reagents are optimized for maximum sensitivity on the interactome and minimal contamination for the mass spectrometer.