Dengue Envelope Domain Ⅲ-Peptide Binding Analysis via Tryptophan Fluorescence Quenching Assay

DOI Web Site Web Site PubMed 参考文献19件
  • Baharuddin Aida
    Drug Design and Development Research Group, University of Malaya Department of Molecular Medicine, Faculty of Medicine, University of Malaya
  • Amir Hassan Asfarina
    Drug Design and Development Research Group, University of Malaya Department of Pharmacy, Faculty of Medicine, University of Malaya
  • Othman Rozana
    Drug Design and Development Research Group, University of Malaya Department of Pharmacy, Faculty of Medicine, University of Malaya Centre for Natural Product and Drug Discovery, University of Malaya
  • Xu Yongtao
    School of Chemistry and Chemical Engineering, Queen’s University Belfast
  • Huang Meilan
    School of Chemistry and Chemical Engineering, Queen’s University Belfast
  • Ario Tejo Bimo
    Center for Infectious Diseases Research, Surya University
  • Yusof Rohana
    Drug Design and Development Research Group, University of Malaya Department of Molecular Medicine, Faculty of Medicine, University of Malaya
  • Abdul Rahman Noorsaadah
    Department of Chemistry, Faculty of Science, University of Malaya
  • Othman Shatrah
    Drug Design and Development Research Group, University of Malaya Department of Molecular Medicine, Faculty of Medicine, University of Malaya

書誌事項

タイトル別名
  • Dengue Envelope Domain III-Peptide Binding Analysis <i>via</i> Tryptophan Fluorescence Quenching Assay

この論文をさがす

抄録

In the efforts to find an anti-viral treatment for dengue, a simple tryptophan fluorescence-screening assay aimed at identifying dengue domain III envelope (EIII) protein inhibitors was developed. Residue Trp391 of EIII was used as an intrinsic probe to monitor the change in fluorescence of the tryptophan residue upon binding to a peptide. The analysis was based on the electron excitation at 280 nm and fluorescence emission at 300–400 nm of EIII, followed by quenching of fluorescence in the presence of potential peptidic inhibitors coded DS36wt, DS36opt, DN58wt and DN58opt. The present study found that the fluorescence of the recombinant EIII was quenched following the binding of DS36opt, DN58wt and DN58opt in a concentration-dependent manner. Since the λmax for emission remained unchanged, the effect was not due to a change in the environment of the tryptophan side chain. In contrast, a minimal fluorescence-quenching effect of DS36wt at 20 and 40 µM suggested that the DS36wt does not have any binding ability to EIII. This was supported by a simple native-page gel retardation assay that showed a band shift of EIII domain when incubated with DS36opt, DN58wt and DN58opt but not with DS36wt. We thus developed a low-cost and convenient spectrophotometric binding assay for the analysis of EIII–peptide interactions in a drug screening application.

収録刊行物

参考文献 (19)*注記

もっと見る

キーワード

詳細情報 詳細情報について

問題の指摘

ページトップへ