Rational design of FRET-based ratiometric chemosensors for in vitro and in cell fluorescence analyses of nucleoside polyphosphates

Yasutaka Kurishita, Takahiro Kohira, Akio Ojida, Itaru Hamachi

研究成果: ジャーナルへの寄稿記事

180 引用 (Scopus)

抄録

Ratiometric fluorescence sensing is a useful technique for the precise and quantitative analysis of biological events occurring under complex conditions, such as those inside cells. We report herein the design of new ratiometric chemosensors for nucleoside polyphosphates such as ATP that are based on binding-induced modulation of fluorescence resonance energy transfer (FRET) coupled with a turn-on fluorescence-sensing mechanism. We designed these new FRET-based ratiometric chemosensors by utilizing spectral overlap changes to modulate the FRET efficiency. Introduction of coumarin fluorophores as the FRET donors into a binuclear zinc complex as the FRET acceptor provided the ratiometric chemosensors. These chemosensors exhibited a clear dual-mission signal change upon binding with strong affinity (Kapp ≈ 10 6-107 M-1) to nucleoside polyphosphates in aqueous solution, whereas no detectable emission change was observed with monophosphates and phosphodiester species or various other anions. These chemosensors were used for real-time fluorescence monitoring of enzyme reactions such as saccharide synthesis by glycosyltransferase and phosphorylation by protein kinase, both of which involve nucleoside polyphosphates as substrates. The utility of ratiometric sensing by chemosensors was further demonstrated in a fluorescence-imaging study of the nucleoside polyphosphates inside living cells, wherein we ratiometrically visualized the stimulus-responsive concentration change of ATP, an indicator of the cellular energy level.

元の言語英語
ページ(範囲)13290-13299
ページ数10
ジャーナルJournal of the American Chemical Society
132
発行部数38
DOI
出版物ステータス出版済み - 9 29 2010

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Polyphosphates
Fluorescence Resonance Energy Transfer
Nucleosides
Fluorescence
Adenosinetriphosphate
Adenosine Triphosphate
Glycosyltransferases
Phosphorylation
Fluorophores
Optical Imaging
Protein Kinases
Electron energy levels
Anions
Zinc
Negative ions
Enzymes
Cells
Modulation
In Vitro Techniques
Proteins

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

これを引用

Rational design of FRET-based ratiometric chemosensors for in vitro and in cell fluorescence analyses of nucleoside polyphosphates. / Kurishita, Yasutaka; Kohira, Takahiro; Ojida, Akio; Hamachi, Itaru.

:: Journal of the American Chemical Society, 巻 132, 番号 38, 29.09.2010, p. 13290-13299.

研究成果: ジャーナルへの寄稿記事

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