Global profiling of protein-DNA and protein-nucleosome binding affinities using quantitative mass spectrometry
Global profiling of protein-DNA and protein-nucleosome binding affinities using quantitative mass spectrometry
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DOI：
10.1038/s41467-018-04084-0
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发表时间：
2018-04-25
影响因子：
16.6
通讯作者：
Vermeulen, Michiel
中科院分区：
文献类型：
--
作者：
Makowski, Matthew M.;Grawe, Cathrin;Vermeulen, Michiel
相互作用蛋白质组学研究为多聚体生物分子组装和细胞尺度分子网络提供了基础性的见解。基于质谱的相互作用蛋白质组学最近的重大发展受到无标记、同位素和同量异位素定量工作流程的快速发展的推动。在这里，我们报告了一个定量的蛋白质-DNA和蛋白质-核小体结合试验，使用亲和纯化从核提取物与同量异位素化学标记和质谱定量表观结合亲和力蛋白质组范围内。我们使用这种测定与各种DNA和核小体诱饵定量单体和多聚体转录因子和染色质重塑复合物的表观结合亲和力。
Interaction proteomics studies have provided fundamental insights into multimeric biomolecular assemblies and cell-scale molecular networks. Significant recent developments in mass spectrometry-based interaction proteomics have been fueled by rapid advances in label-free, isotopic, and isobaric quantitation workflows. Here, we report a quantitative protein-DNA and protein-nucleosome binding assay that uses affinity purifications from nuclear extracts coupled with isobaric chemical labeling and mass spectrometry to quantify apparent binding affinities proteome-wide. We use this assay with a variety of DNA and nucleosome baits to quantify apparent binding affinities of monomeric and multimeric transcription factors and chromatin remodeling complexes.