Howe P W, Allain F H, Varani G, Neuhaus D
MRC Laboratory of Molecular Biology, Cambridge, U.K.
J Biomol NMR. 1998 Jan;11(1):59-84. doi: 10.1023/a:1008297502874.
RNA-protein recognition is critical to post-transcriptional regulation of gene expression, yet poorly understood at the molecular level. The relatively slow progress in understanding this important area of molecular biology is due to difficulties in obtaining good-quality crystals and derivatives, and in preparing samples suitable for NMR investigation. The determination of the structure of the complex between the human U1A protein and its polyadenylation inhibition element is described here. In this paper, we describe the sample preparation, spectral assignments, construction of the NOE-based distance constraints and methodology for calculating the structure of the complex. The structure was determined to an overall precision of 2.03 A (for all ordered regions), and 1.08 A for the protein-RNA interface. The patterns of hydrogen bonding and hydrophobic interactions at the interface were analysed statistically using the final ensemble of 31 structures.
RNA与蛋白质的识别对于基因表达的转录后调控至关重要，但在分子水平上却了解甚少。在理解这一分子生物学重要领域方面进展相对缓慢，原因在于难以获得高质量的晶体和衍生物，以及制备适合核磁共振研究的样品。本文描述了人U1A蛋白与其聚腺苷酸化抑制元件之间复合物结构的测定。在本文中，我们描述了样品制备、光谱归属、基于核Overhauser效应（NOE）的距离限制的构建以及计算该复合物结构的方法。该结构的整体精度确定为2.03埃（对于所有有序区域），蛋白质-RNA界面的精度为1.08埃。使用31个结构的最终集合对界面处的氢键和疏水相互作用模式进行了统计分析。
