Structure-activity studies on the fluorescent indicator in a displacement assay for the screening of small molecules binding to RNA

Shiori Umemoto, Seongwang Im, Jinhua Zhang, Masaki Hagihara, Asako Murata, Yasue Harada, Takeo Fukuzumi, Takahiro Wazaki, Shin Ichi Sasaoka, Kazuhiko Nakatani

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26 Citations (Scopus)

Abstract

A series of xanthone and thioxanthone derivatives with aminoalkoxy substituents were synthesized as fluorescent indicators for a displacement assay in the study of small-molecule-RNA interactions. The RNA-binding properties of these molecules were investigated in terms of the improved binding selectivity to the loop region in the RNA secondary structure relative to 2,7-bis(2-aminoethoxy)xanthone (X2S) by fluorimetric titration and displacement assay. An 11-mer double-stranded RNA and a hairpin RNA mimicking the stem loop IIB of Rev response element (RRE) RNA of HIV-1 mRNA were used. The X2S derivatives with longer aminoalkyl substituents showed a higher affinity to the double-stranded RNA than the parent molecule. Introduction of a methyl group on the aminoethoxy moiety of X2S effectively modulated the selectivity to the RNA secondary structure. Methyl group substitution at the C1' position suppressed the binding to the loop regions. Substitution with two methyl groups on the amino nitrogen atom resulted in reducing the affinity to the double-stranded region by a factor of 40 %. The effect of methyl substitution on the amino nitrogen atom was also observed for a thioxanthone derivative. Titration experiments, however, suggested that thioxanthone derivatives showed a more prominent tendency of multiple binding to RNA than xanthone derivatives. The selectivity index calculated from the affinity to the double-stranded and loop regions suggested that the N,N-dimethyl derivative of X2S would be suitable for the screening of small molecules binding to RRE. Indicating RNA binding: A fluorescent indicator displacement assay using 2,7-bis(aminoethoxy)xanthone (X2S) as the indicator is reported for the study of small-molecule-RNA interactions (see figure). The substituents at the two alkoxy moieties modulate the binding selectivity. The X2S derivative with N,N-dimethylaminoethoxy substituents is the best fluorescent indicator in the screening of molecules binding to Rev response element (RRE).

Original languageEnglish
Pages (from-to)9999-10008
Number of pages10
JournalChemistry - A European Journal
Volume18
Issue number32
DOIs
Publication statusPublished - Aug 6 2012
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Organic Chemistry

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