A modular strategy to construct functional RNA-protein complexes

核糖核肽(RNP)是一种稳定的多肽和RNA的复合体，为构建小分子的大分子受体提供了一个新的框架。RRE RNA和REV肽形成结构特征良好的稳定的RNP复合体，适合于逐步功能化。在体外选择来自基于RRE的RNA文库和REV多肽的RNP池提供了RNP受体，对三磷酸核苷酸、定义的氨基酸序列中的磷酸酪氨酸残基和多巴胺具有特异性。通过生化和核磁共振研究相结合的方法表征了ATP结合RNP的一个新的ATP结合基序。由荧光团标记的REV多肽功能化的RNP受体施加与配体结合事件相关的光学信号。用荧光团修饰的REV多肽取代与ATP结合的RNP的REV多肽，得到了一系列荧光的ATP传感器。这种产生定制荧光传感器的策略被应用于选择性地检测定义的氨基酸序列中特定的磷酸化酪氨酸残基。

A stable complex of a peptide and RNA, ribonucleopeptide(RNP), provides a new framework to construct a macromolecular receptor for small molecules. The RRE RNA and the Rev peptide form a structurally well-characterized stable RNP complex that is suitable for a stepwise functionalization. In vitro selection of the RNP pool originating from an RRE-based RNA library and the Rev peptide affords RNP receptors specific for nucleotide triphosphates, for a phosphotyrosine residue in defined amino acid sequence, and for dopamine. A new ATP-binding motif for the ATP-binding RNP was characterized by a combination of biochemical and NMR studies. The RNP receptor functionalized by a fluorophore-labeled Rev peptide exerts optical signals associated with the ligand binding events. Replacing the Rev peptide of the ATP-binding RNP with a fluorophore-modified Rev peptide affords a series of fluorescent ATP sensors. This strategy to generate tailor-made fluorescent sensors is applied for a selective detection of a specific phosphorylated tyrosine residue within a defined amino acid sequence.