Slupsky C M, Gentile L N, McIntosh L P
Department of Biochemistry and Molecular Biology, and Protein Engineering Network of Centres of Excellence, University of British Columbia, Vancouver, Canada.
Biochem Cell Biol. 1998;76(2-3):379-90. doi: 10.1139/bcb-76-2-3-379.
The measurement of interproton nuclear Overhauser enhancements (NOEs) and dihedral angle restraints of aromatic amino acids is a critical step towards determining the structure of a protein. The complete assignment of the resonances from aromatic rings and the subsequent resolution and identification of their associated NOEs, however, can be a difficult task. Shown here is a strategy for assigning the 1H, 13C, and 15N signals from the aromatic side chains of histidine, tryptophan, tyrosine, and phenylalanine using a suite of homo- and hetero-nuclear scalar and NOE correlation experiments, as well as selective deuterium isotope labelling. In addition, a comparison of NOE information obtained from homonuclear NOE spectroscopy (NOESY) and 13C-edited NOESY-heteronuclear single quantum correlation experiments indicates that high-resolution homonuclear two-dimensional NOESY spectra of selectively deuterated proteins are invaluable for obtaining distance restraints to the aromatic residues.
测量质子间的核Overhauser增强效应（NOE）以及芳香族氨基酸的二面角限制是确定蛋白质结构的关键步骤。然而，对芳香环共振进行完整归属以及随后解析和识别其相关的NOE可能是一项艰巨的任务。本文展示了一种策略，即使用一系列同核和异核标量及NOE相关实验，以及选择性氘同位素标记，来归属组氨酸、色氨酸、酪氨酸和苯丙氨酸芳香侧链的1H、13C和15N信号。此外，对同核NOE光谱（NOESY）和13C编辑的NOESY - 异核单量子相关实验获得的NOE信息进行比较表明，选择性氘代蛋白质的高分辨率同核二维NOESY光谱对于获取与芳香族残基的距离限制非常宝贵。
