Zuiderweg E R, Fesik S W
Pharmaceutical Discovery Division, Abbott Laboratories, Abbott Park, Illinois 60064.
Biochemistry. 1989 Mar 21;28(6):2387-91. doi: 10.1021/bi00432a008.
The utility of three-dimensional heteronuclear NMR spectroscopy for the assignment of 1H and 15N resonances of the inflammatory protein C5a (MW 8500), uniformly labeled with 15N, is demonstrated at a protein concentration of 0.7 mM. It is shown that dramatic simplification of the 2D nuclear Overhauser effect spectrum (NOESY) is obtained by editing with respect to the frequency of the 15N heteronucleus in a third dimension. The improved resolution in the 3D experiment largely facilitates the assignment of protein NMR spectra and allows for the determination of distance constraints from otherwise overlapping NOE cross peaks for purposes of 3D structure determination. The results show that 15N heteronuclear 3D NMR can facilitate the structure determination of small proteins and promises to be a useful tool for the study of larger systems that cannot be studied by conventional 2D NMR techniques.
在蛋白质浓度为0.7 mM的条件下，证明了三维异核核磁共振光谱法对于均匀标记15N的炎症蛋白C5a（分子量8500）的1H和15N共振峰归属的实用性。结果表明，通过在第三维中根据15N异核的频率进行编辑，可以显著简化二维核Overhauser效应谱（NOESY）。三维实验中分辨率的提高极大地促进了蛋白质核磁共振谱的归属，并允许从原本重叠的NOE交叉峰确定距离约束，以用于三维结构测定。结果表明，15N异核三维核磁共振可以促进小蛋白质的结构测定，并有望成为研究传统二维核磁共振技术无法研究的更大系统的有用工具。
