Gardner K H, Kay L E
Protein Engineering Network Centres of Excellence, University of Toronto, Ontario, Canada.
Annu Rev Biophys Biomol Struct. 1998;27:357-406. doi: 10.1146/annurev.biophys.27.1.357.
During the past thirty years, deuterium labeling has been used to improve the resolution and sensitivity of protein NMR spectra used in a wide variety of applications. Most recently, the combination of triple resonance experiments and 2H, 13C, 15N labeled samples has been critical to the solution structure determination of several proteins with molecular weights on the order of 30 kDa. Here we review the developments in isotopic labeling strategies, NMR pulse sequences, and structure-determination protocols that have facilitated this advance and hold promise for future NMR-based structural studies of even larger systems. As well, we detail recent progress in the use of solution 2H NMR methods to probe the dynamics of protein sidechains.
在过去三十年中，氘标记已被用于提高蛋白质核磁共振谱的分辨率和灵敏度，这些谱在广泛的应用中使用。最近，三重共振实验与2H、13C、15N标记样品的结合对于分子量约为30 kDa的几种蛋白质的溶液结构测定至关重要。在此，我们回顾了同位素标记策略、核磁共振脉冲序列和结构测定方案的发展，这些发展推动了这一进展，并为未来基于核磁共振的更大系统结构研究带来了希望。此外，我们详细介绍了使用溶液2H核磁共振方法探测蛋白质侧链动力学的最新进展。
