Determining Binding Kinetics of Intrinsically Disordered Proteins by NMR Spectroscopy.
Determining Binding Kinetics of Intrinsically Disordered Proteins by NMR Spectroscopy.
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DOI：
10.1007/978-1-0716-0524-0_34
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Wright PE
中科院分区：
文献类型：
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作者：
Yang K;Arai M;Wright PE
内在无序蛋白（IDP）独特的结构灵活性是其在细胞过程中的多种功能的核心。涉及IDP的蛋白质-蛋白质相互作用通常是瞬时的和动态的。核磁共振光谱法是一种特别有力的工具，可用于表征国内流离失所者的结构倾向、动力学和相互作用。在这里，我们描述的应用程序的Carr-Purcell-Meiboom-Gill（CPMG）弛豫分散实验结合NMR滴定来表征的动力学和内在无序的蛋白质和它们的目标之间的相互作用的机制。我们说明的方法与参考之间的相互作用的人T细胞白血病病毒I型（HTLV-1）碱性亮氨酸拉链蛋白（HBZ）和它的细胞结合伙伴，转录辅激活因子CBP的KIX域的激活域。
The unique structural flexibility of intrinsically disordered proteins (IDPs) is central to their diverse functions in cellular processes. Protein-protein interactions involving IDPs are frequently transient and dynamic in nature. Nuclear magnetic resonance (NMR) spectroscopy is an especially powerful tool for characterizing the structural propensities, dynamics and interactions of IDPs. Here we describe applications of the Carr-Purcell-Meiboom-Gill (CPMG) relaxation dispersion experiment in combination with NMR titrations to characterize the kinetics and mechanisms of interactions between the intrinsically disordered proteins and their targets. We illustrate the method with reference to interactions between the activation domain of the human T-cell leukemia virus type-I (HTLV-1) basic leucine zipper protein (HBZ) and its cellular binding partner, the KIX domain of the transcriptional coactivator CBP.