13C Saturation-Transfer Difference (STD)-NMR Experiments Using INEPT Polarization Transfer
13C Saturation-Transfer Difference (STD)-NMR Experiments Using INEPT Polarization Transfer
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DOI：
10.1007/s00723-019-01182-0
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发表时间：
2020-01
影响因子：
1
通讯作者：
Hui Xu;Daniel G. Lustig;L. Casabianca
中科院分区：
文献类型：
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作者：
Hui Xu;Daniel G. Lustig;L. Casabianca
饱和转移差 (STD)-NMR 光谱已广泛用于筛选与大受体分子结合的潜在配体。 STD-NMR 实验通常基于质子 NMR 光谱，该光谱可能会出现光谱重叠，从而导致基于 STD 的表位作图中信息缺失。二维 STD-NMR 实验可以减轻光谱重叠，但非常耗时。在这里，我们检查了受体分子中质子饱和的可行性，并观察使用极化转移增强的不敏感核（INEPT）脉冲序列将极化从质子转移到碳后对附近碳核的 STD 效应。我们表明，在有利的条件下，1H→13C STD-INEPT 实验可以提供与二维异核实验相似的信息，但所需时间要少得多。当研究配体混合物时，STD-INEPT 实验特别有用，其中质子和 HSQC 光谱中的峰位置发生显着变化，特别是在使用高通量、自动化方法分析数据时。
Saturation-transfer difference (STD)-NMR spectroscopy has been widely used to screen potential ligands for binding to large receptor molecules. The STD-NMR experiment is typically based on a proton NMR spectrum, which can suffer from spectral overlap, leading to missing information in STD-based epitope mapping. Two-dimensional STD-NMR experiments can alleviate spectral overlap, but are time consuming. Here, we examine the feasibility of saturating protons in a receptor molecule and observing the STD effect on nearby carbon nuclei after transferring polarization from protons to carbons using the insensitive nuclei enhanced by polarization transfer (INEPT) pulse sequence. We show that under favorable conditions, a1H→13C STD-INEPT experiment can give information similar to that obtained from a two-dimensional heteronuclear experiment, but in significantly less time. The STD-INEPT experiment could be especially useful when studying mixtures of ligands in which the peak positions in the proton and HSQC spectrum change significantly, and in particular, when using high-throughput, automated methods to analyze the data.