Application of four-dimensional heteronuclear NMR to the structure determination of a uniformly 13C labeled RNA
Application of four-dimensional heteronuclear NMR to the structure determination of a uniformly 13C labeled RNA
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四维异核核磁共振在均匀 13C 标记 RNA 结构测定中的应用
DOI：
10.1021/ja00029a043
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发表时间：
1992
影响因子：
15
通讯作者：
A. Pardi
中科院分区：
文献类型：
--
作者：
E. Nikonowicz;A. Pardi
图 1.(a) 13C 标记的 r (GGCGCUUGCGUC) 2 双链体的 4D HMQCNOE-HMQC 谱图的 H2'-H5' 糖区图。 4D 谱的此-NOE (frf*) 平面对应于/,= 66.3 ppm 和/3= 74.3 ppm 碳频率。该实验在 Varían VXR-500S 光谱仪上于 30°C 下进行，脉冲序列类似于之前描述的 2，并采用 400 毫秒 NOE 混合时间和 0.9 秒循环时间。 4D 实验的第一个和最后一个 HMQC 部分分别针对 205 和 185 Hz 的 VCH 进行了优化；对于 rh t2、r3 和 Z4 演化时间，分别在 2500、3200、2500 和 3200 Hz 的扫描宽度上收集了 8、32、8 和 512 个复点。补充材料中给出了 4D 实验中使用的脉冲序列、采集和处理参数的其他详细信息，(b) 在类似实验条件下采集的未标记 RNA 双链体的 400 毫秒 2D NOE 谱的相同区域的图。
Figure 1.(a) A plot of the H2'-H5'sugar region of the 4D HMQCNOE-HMQC spectrum of the 13C-labeled r (GGCGCUUGCGUC) 2 duplex. This- NOE (frf*) plane of the 4D spectrum corresponds to/,= 66.3 ppm and/3= 74.3 ppm carbon frequencies. The experiment was carried out on a Varían VXR-500S spectrometer at30 C with a pulse sequence similar to that previously described2 and employed a 400-ms NOE mixing time and a 0.9-s recycle time. The first and last HMQC portions of the 4D experiment were optimized for VCHs of 205 and 185 Hz, respectively; 8, 32, 8, and 512 complex points were collected over sweep widths of 2500, 3200, 2500, and 3200 Hz for the rh t2, r3, andZ4 evolution times, respectively. Additional details of the pulse sequence, acquisition, and processing parameters employed in the 4D experiment are given in the supplementary material,(b) A plot of the same region of a 400-ms 2D NOE spectrum of the unlabeled RNA duplex acquired under similar experimental conditions.