NMR assignment methods for the aromatic ring resonances of phenylalanine and tyrosine residues in proteins

被引:39
作者
Torizawa, T
Ono, AM
Terauchi, T
Kainosho, M
机构
[1] Tokyo Metropolitan Univ, JST, CREST, Hachioji, Tokyo 1920397, Japan
[2] Tokyo Metropolitan Univ, Grad Sch Sci, Hachioji, Tokyo 1920397, Japan
关键词
D O I
10.1021/ja051386m
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
The unambiguous assignment of the aromatic ring resonances in proteins has been severely hampered by the inherently poor sensitivities of the currently available methodologies developed for uniformly C-13/N-15-labeled proteins. Especially, the small chemical shift differences between aromatic ring carbons and protons for phenylalanine residues in proteins have prevented the selective observation and unambiguous assignment of each signal. We have solved all of the difficulties due to the tightly coupled spin systems by preparing regio-/stereoselectively C-13/H-2/N-15-labeled phenylalanine (Phe) and tyrosine (Tyr) to avoid the presence of directly connected C-13-H-1 pairs in the aromatic rings. The superiority of the new labeling schemes for the assignment of aromatic ring signals is clearly demonstrated for a 17 kDa calcium binding protein, calmodulin.
引用
收藏
页码:12620 / 12626
页数:7
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