Sugaya Maki, Saito Ryota, Matsumura Yuriko, Harada Kazuo, Katoh Akira
Department of Materials and Life Science, Faculty of Science and Technology, Seikei University, Musashino, Tokyo 180-8633, Japan.
J Pept Sci. 2008 Aug;14(8):978-83. doi: 10.1002/psc.1021.
A simple method for the detection of specific RNA-polypeptide interactions using MALDI-TOF mass spectroscopy is described. Instead of direct observation of the RNA-polypeptide complex, we attempted the indirect observation of the binding event by focusing on the disappearance of the free polypeptide signal upon interaction with RNA. As a result, specific binding of the Rev-response element (RRE) RNA of the HIV with two RRE-binding peptide aptamers, DLA and RLA peptides, as well as the bacteriophage lambda boxB RNA with the lambda N peptide was observed. We also show that specific RNA-binding peptides can be identified from a mixture of peptides with varying RNA-binding affinity, showing that the method could be applied to high-throughput screening from simple peptide libraries. The method described in this study provides a quick and simple method for detecting specific RNA-polypeptide interactions that avoids difficulties associated with direct observation of RNA and RNA-polypeptide complexes, which may find various applications in the analysis of RNA-polypeptide interactions and in the identification of novel RNA-binding polypeptides.
本文描述了一种使用基质辅助激光解吸电离飞行时间质谱（MALDI-TOF质谱）检测特定RNA-多肽相互作用的简单方法。我们没有直接观察RNA-多肽复合物，而是试图通过关注与RNA相互作用时游离多肽信号的消失来间接观察结合事件。结果，观察到HIV的Rev反应元件（RRE）RNA与两种RRE结合肽适配体DLA和RLA肽的特异性结合，以及噬菌体λ boxB RNA与λ N肽的特异性结合。我们还表明，可以从具有不同RNA结合亲和力的肽混合物中鉴定出特异性RNA结合肽，这表明该方法可应用于从简单肽库进行高通量筛选。本研究中描述的方法提供了一种快速简便的检测特定RNA-多肽相互作用的方法，避免了直接观察RNA和RNA-多肽复合物所带来的困难，该方法可能在RNA-多肽相互作用分析和新型RNA结合多肽的鉴定中找到各种应用。
