Gopinath T, Veglia Gianluigi
Department of Biochemistry, Molecular Biology, and Biophysics- University of Minnesota, Minneapolis, MN 55455.
Department of Chemistry- University of Minnesota, Minneapolis, MN 55455.
Annu Rep NMR Spectrosc. 2016;89:103-121. doi: 10.1016/bs.arnmr.2016.04.003. Epub 2016 Jun 7.
Magic angle spinning solid-state NMR (MAS ssNMR) spectroscopy is a powerful method for structure determination of biomacromolecules that are recalcitrant to crystallization (membrane proteins and fibrils). Conventional multidimensional ssNMR methods acquire one experiment at a time. This approach is time consuming and discards orphan (unused) spin operators. Relatively low sensitivity and poor resolution of protein samples require long acquisition times for multidimensional ssNMR experiments. Here, we describe our recent progress in the development of multiple acquisition solid-state NMR methods for protein structure determination. A family of experiments called Polarization Optimized Experiments (POE) were designed, in which we utilized the orphan spin operators that are discarded in classical multidimensional NMR experiments, recovering them to allow simultaneous acquisition of multiple 2D and 3D experiments, all while using conventional probes with spectrometers equipped with one receiver. Three strategies namely, DUMAS, MEIOSIS, and MAeSTOSO were used for the concatenation of various 2D and 3D experiments. These methods open up new avenues for reducing the acquisition times of multidimensional experiments for biomolecular ssNMR spectroscopy.
魔角旋转固态核磁共振（MAS ssNMR）光谱法是一种用于确定难以结晶的生物大分子（膜蛋白和纤维）结构的强大方法。传统的多维ssNMR方法一次只进行一个实验。这种方法耗时且会丢弃孤立（未使用）的自旋算符。蛋白质样品相对较低的灵敏度和较差的分辨率使得多维ssNMR实验需要较长的采集时间。在此，我们描述了我们在开发用于蛋白质结构测定的多重采集固态核磁共振方法方面的最新进展。设计了一系列名为极化优化实验（POE）的实验，其中我们利用了在经典多维核磁共振实验中被丢弃的孤立自旋算符，将它们回收以允许同时采集多个二维和三维实验，同时使用配备一个接收器的光谱仪的传统探头。三种策略，即DUMAS、减数分裂和MAeSTOSO，用于连接各种二维和三维实验。这些方法为减少生物分子ssNMR光谱多维实验的采集时间开辟了新途径。
