A modular strategy to construct functional RNA-protein complexes

核糖核肽（ribonucleopeptide，RNP）是一种稳定的肽与RNA复合物，为构建小分子的大分子受体提供了新的框架。RRE RNA和Rev肽形成结构良好表征的稳定RNP复合物，其适合于逐步官能化。源自基于RRE的RNA文库和Rev肽的RNP池的体外选择提供了对核苷酸三磷酸、对限定氨基酸序列中的磷酸酪氨酸残基和对多巴胺具有特异性的RNP受体。一个新的ATP结合基序的ATP结合RNP的特点是结合生物化学和NMR研究。由荧光团标记的Rev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肽取代ATP结合RNP的Rev肽提供了一系列荧光ATP传感器。这种生成定制荧光传感器的策略被应用于选择性检测特定氨基酸序列内的特定磷酸化酪氨酸残基。

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.