Sjodt Megan, Clubb Robert T
Department of Chemistry and Biochemistry, UCLA-DOE Institute of Genomics and Proteomics and Molecular Biology Institute, University of California, Los Angeles, USA.
Bio Protoc. 2017 Apr 5;7(7). doi: 10.21769/BioProtoc.2207.
Site-specific attachment of paramagnetic spin labels to biomolecules causes distance-dependent line-broadening effects, which can be exploited to study the structure and dynamics of these molecules in solution. This protocol describes how to attach nitroxide spin labels to proteins and how to collect and analyze NMR data using these labeled samples. We also explain how to derive distance restraints for paramagnetic relaxation enhancement nuclear magnetic resonance (PRE-NMR) studies.
将顺磁自旋标记物位点特异性地附着于生物分子会引起距离依赖性的谱线展宽效应，可利用此效应研究这些分子在溶液中的结构和动力学。本方案描述了如何将氮氧自由基自旋标记物附着于蛋白质，以及如何使用这些标记样品收集和分析核磁共振（NMR）数据。我们还解释了如何为顺磁弛豫增强核磁共振（PRE-NMR）研究推导距离限制。
