Bayerisches NMR Zentrum (BNMRZ) at the Department of Chemistry, Technische Universität München (TUM), Lichtenbergstr. 4, 85747 Garching, Germany.
Helmholtz-Zentrum München (HMGU), Deutsches Forschungszentrum für Gesundheit und Umwelt, Institute of Structural Biology (STB), Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.
Chem Rev. 2022 May 25;122(10):10019-10035. doi: 10.1021/acs.chemrev.1c00681. Epub 2021 Dec 6.
Proton detection developed in the last 20 years as the method of choice to study biomolecules in the solid state. In perdeuterated proteins, proton dipolar interactions are strongly attenuated, which allows yielding of high-resolution proton spectra. Perdeuteration and backsubstitution of exchangeable protons is essential if samples are rotated with MAS rotation frequencies below 60 kHz. Protonated samples can be investigated directly without spin dilution using proton detection methods in case the MAS frequency exceeds 110 kHz. This review summarizes labeling strategies and the spectroscopic methods to perform experiments that yield assignments, quantitative information on structure, and dynamics using perdeuterated samples. Techniques for solvent suppression, H/D exchange, and deuterium spectroscopy are discussed. Finally, experimental and theoretical results that allow estimation of the sensitivity of proton detected experiments as a function of the MAS frequency and the external field in a perdeuterated environment are compiled.
质子检测技术在过去 20 年中得到了发展，成为研究固态生物分子的首选方法。在完全氘代的蛋白质中，质子偶极相互作用被强烈衰减，这允许获得高分辨率的质子谱。如果样品以 MAS 旋转频率低于 60 kHz 进行旋转，则必须对可交换质子进行氘代和回取代。如果 MAS 频率超过 110 kHz，则可以使用质子检测方法直接研究质子化样品，而不会发生自旋稀释。本综述总结了标记策略和光谱方法，用于进行实验，这些实验使用完全氘代的样品提供结构、动态和定量信息的分配。讨论了用于溶剂抑制、H/D 交换和氘谱的技术。最后，编译了实验和理论结果，这些结果允许根据 MAS 频率和外部场在氘化环境中估计质子检测实验的灵敏度。
