Institute of Physical and Theoretical Chemistry, University of Bonn, Wegelerstraße 12, 53115 Bonn, Germany.
EaStCHEM School of Chemistry, Biomedical Sciences Research Complex, and Centre of Magnetic Resonance, University of St Andrews North Haugh, St Andrews KY16 9ST, U.K.
J Am Chem Soc. 2021 Nov 3;143(43):17875-17890. doi: 10.1021/jacs.1c07371. Epub 2021 Oct 19.
Distance distribution information obtained by pulsed dipolar EPR spectroscopy provides an important contribution to many studies in structural biology. Increasingly, such information is used in integrative structural modeling, where it delivers unique restraints on the width of conformational ensembles. In order to ensure reliability of the structural models and of biological conclusions, we herein define quality standards for sample preparation and characterization, for measurements of distributed dipole-dipole couplings between paramagnetic labels, for conversion of the primary time-domain data into distance distributions, for interpreting these distributions, and for reporting results. These guidelines are substantiated by a multi-laboratory benchmark study and by analysis of data sets with known distance distribution ground truth. The study and the guidelines focus on proteins labeled with nitroxides and on double electron-electron resonance (DEER aka PELDOR) measurements and provide suggestions on how to proceed analogously in other cases.
通过脉冲双极电子顺磁共振波谱获得的距离分布信息为结构生物学的许多研究提供了重要贡献。越来越多的此类信息被用于综合结构建模，其中它对构象集的宽度提供了独特的约束。为了确保结构模型和生物学结论的可靠性，我们在此定义了样品制备和表征、测量顺磁标记之间分布偶极-偶极耦合、将主要时域数据转换为距离分布、解释这些分布以及报告结果的质量标准。这些准则通过多实验室基准研究和具有已知距离分布真实值的数据集的分析得到证实。该研究和准则侧重于用氮氧自由基标记的蛋白质和双电子-电子共振（DEER 又名 PELDOR）测量，并就如何在其他情况下类似地进行提供了建议。
