Wenter Philipp, Reymond Luc, Auweter Sigrid D, Allain Frédéric H-T, Pitsch Stefan
Institut des Science et Ingénierie Chimiques, Ecole Polytechnique Fédérale de Lausanne, EPFL-BCH, 1015 Lausanne, Switzerland.
Nucleic Acids Res. 2006 Jun 28;34(11):e79. doi: 10.1093/nar/gkl427.
We report an optimized synthesis of all canonical 2'-O-TOM protected ribonucleoside phosphoramidites and solid supports containing [13C5]-labeled ribose moieties, their sequence-specific introduction into very short RNA sequences and their use for the structure determination of two protein-RNA complexes. These specifically labeled sequences facilitate RNA resonance assignments and are essential to assign a high number of sugar-sugar and intermolecular NOEs, which ultimately improve the precision and accuracy of the resulting structures. This labeling strategy is particularly useful for the study of protein-RNA complexes with single-stranded RNA in solution, which is rapidly an increasingly relevant research area in biology.
我们报道了一种优化合成所有标准的2'-O-TOM保护的核糖核苷亚磷酰胺以及含有[13C5]标记核糖部分的固相载体的方法，将它们序列特异性地引入极短的RNA序列中，并用于确定两种蛋白质-RNA复合物的结构。这些特异性标记的序列有助于RNA共振归属，对于确定大量的糖-糖和分子间核Overhauser效应（NOE）至关重要，这最终提高了所得结构的精度和准确性。这种标记策略对于研究溶液中与单链RNA形成的蛋白质-RNA复合物特别有用，而这在生物学中是一个迅速发展且日益重要的研究领域。
