Tetrathiafulvalene-Labelled Nucleosides and Nucleoside Triphosphates: Synthesis, Electrochemistry and the Scope of Their Polymerase Incorporation into DNA

Authors

  • Jan Riedl,

    1. Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Gilead & IOCB Research Center, Flemingovo nam. 2, 16610 Prague 6, Czech Republic, Fax: +420-220183559
    Search for more papers by this author
  • Petra Horáková,

    1. Institute of Biophysics, Academy of Sciences of the Czech Republic, Kralovopolska 135, 612 65 Brno, Czech Republic, Fax: +420-541211293
    Search for more papers by this author
  • Peter Šebest,

    1. Institute of Biophysics, Academy of Sciences of the Czech Republic, Kralovopolska 135, 612 65 Brno, Czech Republic, Fax: +420-541211293
    Search for more papers by this author
  • Radek Pohl,

    1. Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Gilead & IOCB Research Center, Flemingovo nam. 2, 16610 Prague 6, Czech Republic, Fax: +420-220183559
    Search for more papers by this author
  • Luděk Havran,

    1. Institute of Biophysics, Academy of Sciences of the Czech Republic, Kralovopolska 135, 612 65 Brno, Czech Republic, Fax: +420-541211293
    Search for more papers by this author
  • Miroslav Fojta,

    1. Institute of Biophysics, Academy of Sciences of the Czech Republic, Kralovopolska 135, 612 65 Brno, Czech Republic, Fax: +420-541211293
    Search for more papers by this author
  • Michal Hocek

    1. Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Gilead & IOCB Research Center, Flemingovo nam. 2, 16610 Prague 6, Czech Republic, Fax: +420-220183559
    Search for more papers by this author

Abstract

The title 5-substituted pyrimidines (U and C) and 7-substituted 7-deazapurines (7-deazaA and 7-deazaG) bearing tetrathiafulvelene (TTF) attached through an acetylene linker have been prepared by Sonogashira cross-coupling of the corresponding 5- or 7-iodo derivatives of nucleosides with 2-ethynyltetrathiafulvalene. Their subsequent triphosphorylation gave the corresponding nucleoside triphosphates (dNTPs). Square-wave voltammetry of the TTF-labelled nucleosides and nucleotides showed two peaks, one at 0.2–0.3 V and the other at around 0.65 V (vs. Ag|AgCl|3 M KCl), which correspond to two reversible one-electron redox processes in the TTF moiety. Polymerase incorporation of the TTF-labelled dNTPs into DNA has also been studied. Multiple incorporations were rather problematic and only by using dCTTFTP was efficient primer extension observed with Vent (exo-)polymerase. Single nucleotide extension was successful with labelled A (dA*TTFTP) and C (dCTTFTP) nucleotides. Inhibition of the polymerase was observed at higher concentrations of dNTTFTPs.(© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)

Ancillary