期刊
JOURNAL OF PEPTIDE SCIENCE
卷 14, 期 8, 页码 978-983出版社
JOHN WILEY & SONS LTD
DOI: 10.1002/psc.1021
关键词
RNA-polypeptide interactions; MALDI-TOF mass spectrometry; arginine-rich peptide; HIV RRE; lambda boxB; peptide library
资金
- Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan
- High-Tech Research Center
- MEXT (2004-2008)
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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