Ribo-Proteomics Approach to Profile RNA-Protein and Protein-Protein Interaction Networks.
Ribo-Proteomics Approach to Profile RNA-Protein and Protein-Protein Interaction Networks.
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用于分析 RNA-蛋白质和蛋白质-蛋白质相互作用网络的核糖蛋白质组学方法。
DOI：
10.1007/978-1-4939-3591-8_14
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Yong,Jeongsik
中科院分区：
文献类型：
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作者：
Yeh,Hsin-Sung;Chang,Jae-Woong;Yong,Jeongsik
表征蛋白质-蛋白质和蛋白质- rna相互作用网络是理解rna结合蛋白功能的基本步骤。在许多情况下，这些相互作用是短暂的和高度动态的。因此，捕获活细胞中稳定的和短暂的相互作用，以鉴定蛋白质结合伙伴和绘制rna结合序列是成功建立分子相互作用网络的关键。在本章中，我们将描述一种方法，利用甲醛作为交联剂，从细胞提取物中富集特定的rna -蛋白复合物，然后进行质谱分析和下一代测序分析，以捕获活细胞中的分子相互作用。
Characterizing protein–protein and protein–RNA interaction networks is a fundamental step to understanding the function of an RNA-binding protein. In many cases, these interactions are transient and highly dynamic. Therefore, capturing stable as well as transient interactions in living cells for the identification of protein-binding partners and the mapping of RNA-binding sequences is key to a successful establishment of the molecular interaction network. In this chapter, we will describe a method for capturing the molecular interactions in living cells using formaldehyde as a crosslinker and enriching a specific RNA–protein complex from cell extracts followed by mass spectrometry and Next-Gen sequencing analyses.