Department of Chemistry and Biochemistry, University of Delaware, Newark, Delaware 19716, USA.
J Phys Chem B. 2012 Nov 26;116(46):13585-96. doi: 10.1021/jp3005794. Epub 2012 Nov 12.
We present a time-saving strategy for acquiring 3D magic angle spinning NMR spectra for chemical shift assignments in proteins and protein assemblies in the solid state. By simultaneous application of nonuniform sampling (NUS) and paramagnetic-relaxation-assisted condensed data collection (PACC), we can attain 16-fold time reduction in the 3D experiments without sacrificing the signal-to-noise ratio or the resolution. We demonstrate that with appropriate concentration of paramagnetic dopant introduced into the sample the overwhelming majority of chemical shifts are not perturbed, with the exception of a limited number of shifts corresponding to residues located at the surface of the protein, which exhibit small perturbations. This approach enables multidimensional MAS spectroscopy in samples of intrinsically low sensitivity and/or high spectral congestion where traditional experiments fail, and is especially beneficial for structural and dynamics studies of large proteins and protein assemblies.
我们提出了一种节省时间的策略，用于获取固态下蛋白质和蛋白质组装体的化学位移分配的三维魔角旋转 NMR 谱。通过非均匀采样 (NUS) 和顺磁弛豫辅助浓缩数据采集 (PACC) 的同时应用，我们可以在不牺牲信噪比或分辨率的情况下，将 3D 实验的时间缩短 16 倍。我们证明，在样品中引入适当浓度的顺磁掺杂剂，除了对应于位于蛋白质表面的残基的少数位移外，绝大多数化学位移不会受到干扰，这些残基表现出小的扰动。这种方法使在传统实验失败的固有低灵敏度和/或高光谱拥挤的样品中进行多维 MAS 光谱学成为可能，并且特别有益于大蛋白质和蛋白质组装体的结构和动力学研究。
