Mandal Abhishek, Boatz Jennifer C, Wheeler Travis B, van der Wel Patrick C A
Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15260, USA.
Department of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, 15260, USA.
J Biomol NMR. 2017 Mar;67(3):165-178. doi: 10.1007/s10858-017-0089-6. Epub 2017 Feb 22.
A number of recent advances in the field of magic-angle-spinning (MAS) solid-state NMR have enabled its application to a range of biological systems of ever increasing complexity. To retain biological relevance, these samples are increasingly studied in a hydrated state. At the same time, experimental feasibility requires the sample preparation process to attain a high sample concentration within the final MAS rotor. We discuss these considerations, and how they have led to a number of different approaches to MAS NMR sample preparation. We describe our experience of how custom-made (or commercially available) ultracentrifugal devices can facilitate a simple, fast and reliable sample preparation process. A number of groups have since adopted such tools, in some cases to prepare samples for sedimentation-style MAS NMR experiments. Here we argue for a more widespread adoption of their use for routine MAS NMR sample preparation.
魔角旋转(MAS)固态核磁共振领域的一些最新进展,使其能够应用于越来越复杂的一系列生物系统。为保持生物学相关性,这些样品越来越多地在水合状态下进行研究。与此同时,实验可行性要求样品制备过程在最终的MAS转子内达到高样品浓度。我们讨论了这些考虑因素,以及它们如何导致了多种不同的MAS核磁共振样品制备方法。我们描述了使用定制(或市售)超速离心设备如何能够实现简单、快速且可靠的样品制备过程的经验。此后,许多研究小组采用了此类工具,在某些情况下用于制备沉降式MAS核磁共振实验的样品。在此,我们主张更广泛地将其用于常规MAS核磁共振样品制备。