A five-dimensional (5D) APSY (automated projection spectroscopy) HCNCH experiment is presented, which allows unambiguous correlation of sugar to base nuclei in nucleic acids. The pulse sequence uses multiple quantum (MQ) evolution which enables long constant-time evolution periods in all dimensions, an improvement that can also benefit non-APSY applications. Applied with an RNA with 23 nucleotides the 5D APSY-HCNCH experiment produced a complete and highly precise 5D chemical shift list within 1.5 h. Alternatively, and for molecules where the out-and-stay 5D experiment sensitivity is not sufficient, a set of out-and-back 3D APSY-HCN experiments is proposed: an intra-base (3D APSY-b-HCN) experiment in an MQ or in a TROSY version, and an MQ sugar-to-base (3D APSY-s-HCN) experiment. The two 3D peak lists require subsequent matching via the N1/9 chemical shift values to one 5D peak list. Optimization of the 3D APSY experiments for maximal precision in the N1/9 dimension allowed matching of all (15)N chemical shift values contained in both 3D peak lists. The precise 5D chemical shift correlation lists resulting from the 5D experiment or a pair of 3D experiments also provide a valuable basis for subsequent connection to chemical shifts derived with other experiments.

译文

:提出了一种五维(5D)APSY(自动投影光谱法)HCNCH实验,该实验允许糖与核酸中的碱基核明确相关。脉冲序列使用多重量子(MQ)演化,可以在所有维度上实现长的恒定时间演化周期,这一改进也可以使非APSY应用受益。将5D APSY-HCNCH实验物与具有23个核苷酸的RNA结合使用,可在1.5小时内产生完整且高度精确的5D化学位移列表。替代地,对于不适合5D实验的敏感性的分子,提出了一系列不需3D APSY-HCN的实验:一个碱基内(3D APSY-b-HCN)实验MQ或TROSY版本,以及MQ糖对碱(3D APSY-s-HCN)实验。这两个3D峰列表需要随后通过N1 / 9化学位移值与一个5D峰列表进行匹配。优化3D APSY实验以获得N1 / 9维最大精度的结果,可以匹配两个3D峰列表中包含的所有(15)N化学位移值。由5D实验或一对3D实验产生的精确5D化学位移相关性列表,也为后续与其他实验得出的化学位移的关联提供了有价值的基础。

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