Significantly improved resolution for noe correlations from valine and isoleucine (C(y)2) methyl groups in 15N, 13C- and 15N, 13C, 2H-labeled proteins
Significantly improved resolution for noe correlations from valine and isoleucine (C(y)2) methyl groups in 15N, 13C- and 15N, 13C, 2H-labeled proteins
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显着提高 15N、13C 和 15N、13C、2H 标记蛋白中缬氨酸和异亮氨酸 (C(y)2) 甲基基团的 noe 相关性分辨率
DOI：
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发表时间：
1998
期刊：
影响因子：
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通讯作者：
L. Kay
中科院分区：
文献类型：
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作者：
C. Zwahlen;S. Vincent;K. Gardner;L. Kay
描述了一种新的NMR实验，用于记录15N，13C标记或甲基化，15N,13C， 2h标记的蛋白质中Val和Ile甲基的NOEs，其分辨率远远优于传统的3D 13C编辑NOESY数据集。通过记录Cβ和Cγ (Val)或Cγ2 （Ile）的化学位移以及目标质子的化学位移来实现分辨率，并引入了在Cβ碳磁化的恒定时间演化过程中重新聚焦同核碳偶联的策略。通过对160个残基完全质子化的15N， 13c标记的dNumb PTB结构域-肽复合物和甲基质子化的高度氘化的15N， 13c标记的麦芽糖结合蛋白和β-环糊精的复合物（42 kDa）的应用，证明了该方法的实用性。
A new NMR experiment is described for recording NOEs from Val and Ile methyl groups in 15N,13C-labeled or methyl-protonated, 15N,13C,2H-labeled proteins that offers far superior resolution than conventional 3D 13C-edited NOESY data sets. Resolution is achieved by recording both the Cβ and Cγ (Val) or Cγ2 (Ile) chemical shifts as well as the chemical shift of the destination proton, and a strategy is introduced for refocusing homonuclear carbon couplings during the constant-time evolution of Cβ carbon magnetization. The utility of the method is demonstrated with applications on a 160-residue fully protonated 15N,13C-labeled, dNumb PTB domain−peptide complex and a methyl protonated, highly deuterated 15N,13C-labeled complex of maltose binding protein and β-cyclodextrin (42 kDa).