Structural Biology and NMR Laboratory and the Linderstrøm-Lang Centre for Protein Science, Department of Biology, University of Copenhagen, Copenhagen N, Denmark.
Methods Mol Biol. 2020;2141:195-209. doi: 10.1007/978-1-0716-0524-0_9.
Intrinsically disordered proteins (IDPs) have no single, fixed tertiary structure, yet they take on many vital functions in biology. In recent years, considerable effort has been put into the structural characterization of their conformational ensembles, to understand the link between the transient, short- and long-range organizations of IDPs and their functions. Such biophysical studies require substantial amounts of pure protein, representing a major bottleneck in the studies of IDPs. However, the unique physicochemical properties resulting from their compositional bias may be exploited for simple yet effective purification strategies. In this chapter, we provide tips and tricks for IDP production and describe the most important analyses to carry out before bringing an IDP of interest to the laboratory. We outline four purification protocols utilizing the unique properties of IDPs as well as some commonly encountered challenges and pitfalls.
无定形蛋白质（IDPs）没有单一、固定的三级结构，但它们在生物学中具有许多重要的功能。近年来，人们投入了相当大的努力来对其构象集合体进行结构表征，以了解 IDPs 的瞬态、短程和长程组织与它们功能之间的联系。这类生物物理研究需要大量的纯蛋白质，这是 IDPs 研究的主要瓶颈。然而，它们组成上的偏向所导致的独特物理化学性质可以被用于简单而有效的纯化策略。在本章中，我们提供了一些 IDP 生产的技巧和窍门，并描述了在将感兴趣的 IDP 带到实验室之前需要进行的最重要的分析。我们概述了四个利用 IDPs 独特性质的纯化方案，以及一些常见的挑战和陷阱。
