Jamge Suraj, Angenent Gerco C, Bemer Marian
Laboratory of Molecular Biology, Wageningen University & Research, Droevendaalsesteeg 1, 6708, PB, Wageningen, The Netherlands.
Business Unit Bioscience, Wageningen Plant Research, Wageningen University & Research, Droevendaalsesteeg 1, 6708, PB, Wageningen, The Netherlands.
Methods Mol Biol. 2018;1675:315-329. doi: 10.1007/978-1-4939-7318-7_18.
Gene regulation by transcription factors involves complex protein interaction networks, which include chromatin remodeling and modifying proteins as an integral part. Decoding these protein interactions is crucial for our understanding of chromatin-mediated gene regulation. Here, we describe a method for the immunoprecipitation of in planta nuclear protein complexes followed by mass spectrometry (IP-MS) to identify interactions between transcription factors and chromatin remodelers/modifiers in plants. In addition to a step-by-step bench protocol for immunoprecipitation and subsequent mass spectrometry, we provide guidelines and pointers on necessary controls and data analysis approaches.
转录因子介导的基因调控涉及复杂的蛋白质相互作用网络,其中包括染色质重塑和修饰蛋白,它们是该网络不可或缺的一部分。解析这些蛋白质相互作用对于我们理解染色质介导的基因调控至关重要。在此,我们描述了一种用于在植物体内免疫沉淀核蛋白复合物,随后进行质谱分析(IP-MS)的方法,以鉴定植物中转录因子与染色质重塑因子/修饰因子之间的相互作用。除了免疫沉淀及后续质谱分析的详细实验方案外,我们还提供了关于必要对照和数据分析方法的指南及要点。