Detection of Bioactive Small Molecules by Fluorescent Resonance Energy Transfer (FRET) in RNA−Protein Conjugates

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• 作者：Tamaki Endoh ; Ryo Shintani ; Masayasu Mie ; Eiry Kobatake ; Takashi Ohtsuki ; Masahiko Sisido
• 刊名：Bioconjugate Chemistry
• 出版年：2009
• 出版时间：December 16, 2009
• 年：2009
• 卷：20
• 期：12
• 页码：2242-2246
• 全文大小：222K
• 年卷期：v.20,no.12(December 16, 2009)
• ISSN：1520-4812

文摘

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.