Fluorescent N2,N3-ε-adenine nucleoside and nucleotide probes: Synthesis, spectroscopic properties, and biochemical evaluation

Einat Sharon, Sébastien A. Lévesque, Mercedes N. Munkonda, Jean Sévigny, Denise Ecke, Georg Reiser, Bilha Fischer

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

N1,N6-ethenoadenine, ε-A, nucleos(t)ides have been previously applied as fluorescent probes in numerous biochemical systems. However, these ε-A analogues lack the H-bonding capability of adenine. To improve the fluorescence characteristics while preserving the H-bonding pattern required for molecular recognition, we designed a novel probe: N 2,N3-etheno-adenosine, (N2,N3-ε-A). Here, we describe four novel syntheses of the target ε-nucleoside and related analogues. These methods are short, facile, and provide the product regiospecifically. In addition, we report the absorption and emission spectra of N2,N3- ε-A and the dependence of the spectral features on the pH and polarity of the medium. Specifically, maximum emission of N2,N3-ε-A in water is observed at 420 nm (φ = 0.03, excitation at 290 nm). The biochemical relevance of the new probe was evaluated with respect to the P2Y1 receptor and NTPDases 1 and 2. N2,N3-ε-ATP was found to be almost equipotent with ATP at the P2Y1 receptor and was hydrolyzed by NTPDases 1 and 2 at about 80% of the rate of ATP. Furthermore, protein binding does not seem to shift the fluorescence of N2,N3-ε-ATP. Based on the fluorescence and full recognition by ATP-binding proteins, we propose N2,N3-ε-ATP and related nucleo(s)tides as unique probes for the Investigation of adenine nucleo(s)tide-binding proteins as well as for other biochemical applications.

Original languageEnglish
Pages (from-to)1361-1374
Number of pages14
JournalChemBioChem
Volume7
Issue number9
DOIs
StatePublished - Sep 2006

Keywords

  • Fluorescent probes
  • NTPDase
  • Nucleosides
  • Nucleotides
  • P2Y receptor

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