Speeding up direct (15)N detection: hCaN 2D NMR experiment.
Speeding up direct (15)N detection: hCaN 2D NMR experiment.
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DOI：
10.1007/s10858-011-9580-7
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发表时间：
2011-12
影响因子：
2.7
通讯作者：
Wagner G
中科院分区：
文献类型：
--
作者：
Gal M;Edmonds KA;Milbradt AG;Takeuchi K;Wagner G
最近提出了检测低回旋磁核的实验，利用这些核相对于1H的相对慢的弛豫特性。本文提出了一种新型的15N直接探测实验。与之前提出的CaN实验一样，本文描述的hCaN实验也将酰胺15N共振顺序连接起来，但利用了初始的高极化和更快的1H核恢复来缩短循环延迟。这允许在几个小时内记录二维15N检测到的蛋白质核磁共振实验，同时仍然获得13C和15N的优越分辨率，通过脯氨酸建立序列分配，并在酰胺质子快速交换的条件下。实验在多种生物分子上进行了演示，包括小球状蛋白GB1、eIF4G的22kDa HEAT2结构域，以及包含许多SerPro序列重复的NFAT1的非结构化多肽片段。
Experiments detecting low gyromagnetic nuclei have recently been proposed to utilize the relatively slow relaxation properties of these nuclei in comparison to 1H. Here we present a new type of 15N direct—detection experiment. Like the previously proposed CaN experiment, the hCaN experiment described here sequentially connects amide 15N resonances, but utilizes the initial high polarization and the faster recovery of the 1H nucleus to shorten the recycling delay. This allows recording 2D 15N- detected NMR experiments on proteins within a few hours, while still obtaining superior resolution for 13C and 15N, establishing sequential assignments through prolines, and at conditions where amide protons exchange rapidly. The experiments are demonstrated on various biomolecules, including the small globular protein GB1, the 22kDa HEAT2 domain of eIF4G, and an unstructured polypeptide fragment of NFAT1, which contains many SerPro sequence repeats.
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影响因子：
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作者：
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通讯作者：
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