Methods for sequential resonance assignment in solid, uniformly 13C, 15N labelled peptides:: Quantification and application to antamanide
Methods for sequential resonance assignment in solid, uniformly 13C, 15N labelled peptides:: Quantification and application to antamanide
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DOI：
10.1023/a:1011212100630
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发表时间：
2001-07-01
影响因子：
2.7
通讯作者：
Meier, BH
中科院分区：
文献类型：
--
作者：
Detken, A;Hardy, EH;Meier, BH
对于环状十肽金刚烷胺，证明了绝热偏振转移实验在固体、均匀 C-13-N-15 标记的多肽中共振分配的应用。使用 DREAM 序列进行绝热偶极转移的同核相关实验产生了 C-α 和脂肪族侧链 C-13 共振与氨基酸类型的完整分配。使用最近引入的 P9(12)(1) TOBSY 序列可以从 TOBSY 实验中获得相同的信息，该序列采用 J 耦合作为转移机制。对两种方法进行了比较。除了一些芳香族苯丙氨酸共振之外，antamide 中 C-13 和 N-15 共振的完整序列特异性分配是通过一系列选择性或宽带绝热三重共振实验实现的。通过绝热通道 Hartmann-Hahn 交叉极化进行的异核转移与通过 DREAM 进行的绝热同核转移以及旋转共振挠痒序列结合到二维和三维实验中。这些实验的性能是定量评估的。
The application of adiabatic polarization-transfer experiments to resonance assignment in solid, uniformly C-13-N-15-labelled polypeptides is demonstrated for the cyclic decapeptide antamanide. A homonuclear correlation experiment employing the DREAM sequence for adiabatic dipolar transfer yields a complete assignment of the C-alpha and aliphatic side-chain C-13 resonances to amino acid types. The same information can be obtained from a TOBSY experiment using the recently introduced P9(12)(1) TOBSY sequence, which employs the J couplings as a transfer mechanism. A comparison of the two methods is presented. Except for some aromatic phenylalanine resonances, a complete sequence-specific assignment of the C-13 and N-15 resonances in antamanide is achieved by a series of selective or broadband adiabatic triple-resonance experiments. Heteronuclear transfer by adiabatic-passage Hartmann-Hahn cross polarization is combined with adiabatic homonuclear transfer by the DREAM and rotational-resonance tickling sequences into two- and three-dimensional experiments. The performance of these experiments is evaluated quantitatively.