PDF(Pubmed)
Abstract：
Reactive RNA probes are useful for studying and identifying RNA-binding proteins. To that end, we designed and synthesized chloroacetamide-linked 7-deaza-ATP which was a good substrate for T7 RNA polymerase in in vitro transcription assay to synthesize reactive RNA probes bearing one or several reactive modifications. Modified RNA probes reacted with thiol-containing molecules as well as with cysteine- or histidine-containing peptides to form stable covalent products. They also reacted selectively with RNA-binding proteins to form cross-linked conjugates in high conversions thanks to proximity effect. Our modified nucleotide and RNA probes are promising tools for applications in RNA (bio)conjugations or RNA proteomics.
摘要：
反应性RNA探针可用于研究和鉴定RNA结合蛋白。为此,我们设计并合成了氯乙酰胺连接的7-deaza-ATP，它是体外转录测定中T7RNA聚合酶的良好底物，可以合成带有一个或几个反应性修饰的反应性RNA探针。修饰的RNA探针与含巯基分子以及含半胱氨酸或组氨酸的肽反应以形成稳定的共价产物。由于邻近效应,它们还选择性地与RNA结合蛋白反应以形成高转化率的交联缀合物。我们修饰的核苷酸和RNA探针是应用于RNA（生物）缀合或RNA蛋白质组学的有前途的工具。
