Department of Chemistry, and Interdisciplinary Nanoscience Center (iNANO), Aarhus University 8000, Aarhus, Denmark.
Methods Mol Biol. 2020;2141:337-344. doi: 10.1007/978-1-0716-0524-0_17.
Determining hydrogen exchange kinetics in proteins can shed light on their structure and dynamics. Nuclear magnetic resonance (NMR) spectroscopy is an important analytical technique to determine exchange rates. In this chapter, we describe a new method (Paris-DÉCOR) to determine fast protein amide backbone hydrogen exchange rates in the range 10 to 10 s. Measuring fast exchange rates is particularly important for the study of intrinsically disordered proteins, where there is very little protection from exchange to the solvent by the formation of persistent structure. We provide a protocol to set up the experiment as well as MATLAB scripts for numerical simulation that is needed to determine the exchange rates.
确定蛋白质中的氢交换动力学可以揭示它们的结构和动态。核磁共振(NMR)光谱是确定交换速率的重要分析技术。在本章中,我们描述了一种新方法(Paris-DÉCOR),用于确定 10 到 10 s 范围内的快速蛋白质酰胺骨架氢交换速率。测量快速交换速率对于研究固有无序蛋白质特别重要,因为在形成持久结构的情况下,很少有保护不受溶剂交换的影响。我们提供了一个实验方案以及 MATLAB 脚本,用于数值模拟以确定交换速率。