High-throughput screening assay of hepatitis C virus helicase inhibitors using fluorescence-quenching phenomenon

Hidenori Tani, Nobuyoshi Akimitsu, Osamu Fujita, Yasuyoshi Matsuda, Ryo Miyata, Satoshi Tsuneda, Masayuki Igarashi, Yuji Sekiguchi, Naohiro Noda*

*この研究の対応する著者

研究成果: Article査読

19 被引用数 (Scopus)

抄録

We have developed a novel high-throughput screening assay of hepatitis C virus (HCV) nonstructural protein 3 (NS3) helicase inhibitors using the fluorescence-quenching phenomenon via photoinduced electron transfer between fluorescent dyes and guanine bases. We prepared double-stranded DNA (dsDNA) with a 5′-fluorescent-dye (BODIPY FL)-labeled strand hybridized with a complementary strand, the 3′-end of which has guanine bases. When dsDNA is unwound by helicase, the dye emits fluorescence owing to its release from the guanine bases. Our results demonstrate that this assay is suitable for quantitative assay of HCV NS3 helicase activity and useful for high-throughput screening for inhibitors. Furthermore, we applied this assay to the screening for NS3 helicase inhibitors from cell extracts of microorganisms, and found several cell extracts containing potential inhibitors.

本文言語English
ページ(範囲)1054-1059
ページ数6
ジャーナルBiochemical and Biophysical Research Communications
379
4
DOI
出版ステータスPublished - 2009 2 20

ASJC Scopus subject areas

  • 生物理学
  • 生化学
  • 分子生物学
  • 細胞生物学

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