Joint x-ray and NMR refinement of the yeast L30e-mRNA complex
Joint x-ray and NMR refinement of the yeast L30e-mRNA complex
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DOI：
10.1016/j.str.2004.04.023
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发表时间：
2004-07-01
期刊：
影响因子：
5.7
通讯作者：
Williamson, JR
中科院分区：
文献类型：
--
作者：
Chao, JA;Williamson, JR
L30 e是酿酒酵母核糖体蛋白，通过与位于其prem-RNA和成熟mRNA中的富含嘌呤的不对称内环结合来调节其自身的表达。在3.24埃下解析与含有前mRNA调节位点的RNA复合的MBP-L30 e融合蛋白的晶体结构。有趣的是，RNA的结构与先前确定的复合物NMR结构中观察到的不同。NMR数据的分析导致在内环中鉴定出单个亚氨基质子共振，其被错误地分配，并且是错误的RNA结构的主要原因。使用X射线衍射数据和NMR衍生的距离和角度限制进行结构细化。联合NMR和X-射线细化导致改进的立体化学和较低的晶体学R因子。MBP-L30 e-mRNA复合物的RNA内环采用典型的K-转角折叠。
L30e, a Saccharomyces cervisiae ribosomal protein, regulates its own expression by binding to a purine-rich asymmetric internal loop located in both its prem-RNA and mature mRNA. A crystal structure of an MBP-L30e fusion protein in complex with an RNA containing the pre-mRNA regulatory site was solved at 3.24 Angstrom. Interestingly, the structure of the RNA differed from that observed in a previously determined NMR structure of the complex. Analysis of the NMR data led to the identification of a single imino proton resonance in the internal loop that had been incorrectly assigned and was principally responsible for the erroneous RNA structure. A structure refinement was performed using both the X-ray diffraction data and the NMR-derived distance and angle restraints. The joint NMR and X-ray refinement resulted in improved stereochemistry and lower crystallographic R factors. The RNA internal loop of the MBP-L30e-mRNA complex adopts the canonical K-turn fold.