Facile detection of specific RNA-polypeptide interactions by MALDI-TOF mass spectrometry. |
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Authors: | Maki Sugaya Ryota Saito Yuriko Matsumura Kazuo Harada Akira Katoh |
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Institution: | Department of Materials and Life Science, Faculty of Science and Technology, Seikei University, Musashino, Tokyo 180-8633, Japan. |
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Abstract: | 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. Copyright (c) 2008 European Peptide Society and John Wiley & Sons, Ltd. |
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Keywords: | RNA–polypeptide interactions MALDI‐TOF mass spectrometry arginine‐rich peptide HIV RRE lambda boxB peptide library |
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