VIB Department of Structural Biology, Vrije Universiteit Brussel, Brussels 1050, Belgium.
Molecules. 2013 Sep 4;18(9):10802-28. doi: 10.3390/molecules180910802.
Recent advances in NMR methodology and techniques allow the structural investigation of biomolecules of increasing size with atomic resolution. NMR spectroscopy is especially well-suited for the study of intrinsically disordered proteins (IDPs) and intrinsically disordered regions (IDRs) which are in general highly flexible and do not have a well-defined secondary or tertiary structure under functional conditions. In the last decade, the important role of IDPs in many essential cellular processes has become more evident as the lack of a stable tertiary structure of many protagonists in signal transduction, transcription regulation and cell-cycle regulation has been discovered. The growing demand for structural data of IDPs required the development and adaption of methods such as 13C-direct detected experiments, paramagnetic relaxation enhancements (PREs) or residual dipolar couplings (RDCs) for the study of 'unstructured' molecules in vitro and in-cell. The information obtained by NMR can be processed with novel computational tools to generate conformational ensembles that visualize the conformations IDPs sample under functional conditions. Here, we address NMR experiments and strategies that enable the generation of detailed structural models of IDPs.
近年来，NMR 方法和技术的进步使得能够以原子分辨率研究越来越大的生物分子的结构。NMR 光谱特别适用于研究天然无序蛋白质 (IDPs) 和天然无序区域 (IDRs)，这些蛋白质和区域通常非常灵活，在功能条件下没有明确的二级或三级结构。在过去的十年中，由于发现信号转导、转录调控和细胞周期调控中的许多主角缺乏稳定的三级结构，IDPs 在许多重要细胞过程中的重要作用变得更加明显。对 IDPs 结构数据的需求不断增长，要求开发和适应一些方法，如 13C-直接检测实验、顺磁松弛增强 (PREs) 或残差偶极耦合 (RDCs)，以便在体外和细胞内研究“无结构”分子。通过 NMR 获得的信息可以用新的计算工具进行处理，以生成构象集合，直观地展示 IDPs 在功能条件下采样的构象。在这里，我们讨论了能够生成 IDPs 详细结构模型的 NMR 实验和策略。
