NMR Spectroscopic Studies of the Conformational Ensembles of Intrinsically Disordered Proteins
NMR Spectroscopic Studies of the Conformational Ensembles of Intrinsically Disordered Proteins
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DOI：
10.1007/978-3-319-20164-1_5
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发表时间：
2015-01-01
期刊：
影响因子：
--
通讯作者：
Konrat, Robert
中科院分区：
文献类型：
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作者：
Kurzbach, Dennis;Kontaxis, Georg;Konrat, Robert
内含子无序蛋白（IDP）的特点是大量的构象灵活性，因此不服从传统的结构生物学技术。鉴于其固有的结构灵活性，NMR光谱为IDP的结构和动态研究提供了独特的机会。在过去的二十年里，NMR光谱学取得了重大发展，将自旋物理和化学的进步与广泛的应用结合起来。本章将总结NMR方法的主要进展。尽管有效的（多维）NMR实验的信号分配的IDPs的可用性进行了讨论，NMR的更大，更复杂的IDPs需要利用特定的同位素标记策略的光谱简化策略。原型应用同位素标记策略进行了说明。由于IDP-配体缔合和解离过程经常发生在适合NMR光谱的时间尺度上，我们详细描述了CPMG弛豫分散技术在IDP蛋白结合研究中的应用。最后，我们证明了NMR和EPR数据的互补使用提供了一个更全面的图片的构象状态的IDPs，可以用来分析IDPs的构象合奏。
Intrinsically disordered proteins (IDPs) are characterized by substantial conformational flexibility and thus not amenable to conventional structural biology techniques. Given their inherent structural flexibility NMR spectroscopy offers unique opportunities for structural and dynamic studies of IDPs. The past two decades have witnessed significant development of NMR spectroscopy that couples advances in spin physics and chemistry with a broad range of applications. This chapter will summarize key advances in NMR methodology. Despite the availability of efficient (multi-dimensional) NMR experiments for signal assignment of IDPs it is discussed that NMR of larger and more complex IDPs demands spectral simplification strategies capitalizing on specific isotope-labeling strategies. Prototypical applications of isotope labeling-strategies are described. Since IDP-ligand association and dissociation processes frequently occur on time scales that are amenable to NMR spectroscopy we describe in detail the application of CPMG relaxation dispersion techniques to studies of IDP protein binding. Finally, we demonstrate that the complementary usage of NMR and EPR data provide a more comprehensive picture about the conformational states of IDPs and can be employed to analyze the conformational ensembles of IDPs.