Detection of bioactive small molecules by fluorescent resonance energy transfer (FRET) in RNA-protein conjugates

Tamaki Endoh, Ryo Shintani, Masayasu Mie, Eiry Kobatake, Takashi Ohtsuki, Masahiko Sisido

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Bioactive small molecules such as metabolites and drugs play important roles in regulating biological functions. A technique for visualizing such small molecules is very useful to understand their molecular mechanisms. In this study, an RNA-protein conjugate, which consists of an RRE-RNA sensor protein (EYFP-Rev-ECFP) and an altered RRE-RNA, was constructed to detect bioactive small molecules by fluorescent resonance energy transfer (FRET). We designed a theophylline-aptamer-inserted RRE-RNA (Theo-RRE) to detect theophylline as a model target molecule. Theo-RRE formed an RNA-protein conjugate with EYFP-Rev-ECFP in the presence of theophylline. As a result, theophylline was specifically detected down to 10 μM by the FRET increase in distinction from theophylline analogue, caffeine, in cell lysates.

Original languageEnglish
Pages (from-to)2242-2246
Number of pages5
JournalBioconjugate Chemistry
Volume20
Issue number12
DOIs
Publication statusPublished - Dec 16 2009

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Energy Transfer
Theophylline
Transfer RNA
RNA
Energy transfer
Proteins
Molecules
rev Gene Products
Caffeine
Metabolites
Sensors
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Organic Chemistry
  • Pharmaceutical Science
  • Biomedical Engineering
  • Pharmacology

Cite this

Detection of bioactive small molecules by fluorescent resonance energy transfer (FRET) in RNA-protein conjugates. / Endoh, Tamaki; Shintani, Ryo; Mie, Masayasu; Kobatake, Eiry; Ohtsuki, Takashi; Sisido, Masahiko.

In: Bioconjugate Chemistry, Vol. 20, No. 12, 16.12.2009, p. 2242-2246.

Research output: Contribution to journalArticle

Endoh, Tamaki ; Shintani, Ryo ; Mie, Masayasu ; Kobatake, Eiry ; Ohtsuki, Takashi ; Sisido, Masahiko. / Detection of bioactive small molecules by fluorescent resonance energy transfer (FRET) in RNA-protein conjugates. In: Bioconjugate Chemistry. 2009 ; Vol. 20, No. 12. pp. 2242-2246.
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