A practical implementation of cross-spectrum in protein backbone resonance assignment
A practical implementation of cross-spectrum in protein backbone resonance assignment
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DOI：
10.1016/j.jmr.2009.12.018
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发表时间：
2010-04-01
影响因子：
2.2
通讯作者：
Tjandra, Nico
中科院分区：
文献类型：
--
作者：
Chen, Kang;Delaglio, Frank;Tjandra, Nico
将交叉谱的概念应用到蛋白质核磁共振谱中，以辅助主干的顺序共振指认。互谱分析通常用于揭示频域中的相关性，作为揭示包含在多个时间序列中的共同特征的一种手段。在这里，相关的核磁共振谱之间的交叉光谱，例如HNCO和HN(CA)CO，可以沿着它们共同的C‘碳轴通过逐点相乘来计算。在得到的高阶互谱中，在每个共同的I-1碳频率上都会出现增强的相关信号，从而能够识别来自残基I和I-1的酰胺质子H-N(和氮N)共振。在N-15/C-13双标记泛素样品上测量了H(N)CO、H(NCA)CO、H(NCO)CACB和H(N)CACB的2D光谱。这些2D谱被用来计算两个准3D交叉谱，H-I-Hi-1-C‘(I-1)和H-I-Hi-1-CA(I-1)CB(I-1)。我们发现，使用这种方法，H，C‘，CA和CB的主链共振可以被完全指定，而不会有歧义。互谱原理有望为异核主链共振的归属提供一种简单、实用、更定量的方法。由爱思唯尔公司出版。
The concept of cross-spectrum is applied in protein NMR spectroscopy to assist in the backbone sequential resonance assignment. Cross-spectrum analysis is used routinely to reveal correlations in frequency domains as a means to reveal common features contained in multiple time series. Here the cross-spectrum between related NMR spectra, for example HNCO and HN(CA)CO, can be calculated with point-by-point multiplications along their common C' carbon axis. In the resulting higher order cross-spectrum, an enhanced correlation signal occurs at every common i-1 carbon frequency allowing the amide proton H-N (and nitrogen N) resonances from residues i and i-1 to be identified. The cross-spectrum approach is demonstrated using 2D spectra H(N)CO, H(NCA)CO, H(NCO)CACB, and H(N)CACB measured on a N-15/C-13 double-labeled Ubiquitin sample. These 2D spectra are used to calculate two pseudo-3D cross-spectra, H-i-Hi-1-C'(i-1) and H-i-Hi-1-CA(i-1)CB(i-1). We show using this approach, backbone resonances of H, C', CA, and CB can be fully assigned without ambiguity. The cross-spectrum principle is expected to offer an easy, practical, and more quantitative approach for heteronuclear backbone resonance assignment. Published by Elsevier Inc.