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The effect of divalent cations on the catalytic activity of the human plasma 3′-exonuclease

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Abstract

The 3′-exonuclease from human plasma is a soluble form of nucleotide pyrophosphatase/phosphodiesterase 1 (NPP1) (EC 3.1.4.1/EC 3.6.1.9). Here, the possibility of divalent cation influence for the 3′-exonuclease activity was investigated using the phosphorothioate congener of oligonucleotide containing all phosphorothioate internucleotide linkages of the [RP]-configuration ([RP-PS]-d[T12]) as the substrate for this enzyme. It was found that the 3′-exonuclease is a metalloenzyme, i.e. its phosphodiesterase activity was completely abolished at 0.8 mM concentration EDTA and, in turn, it was restored in the presence of Mg2+ or Mn2+ ions. In addition, Mg2+ can be replaced effectively by Ca2+, Mn2+, or Co2+, but not by Ni2+ and Cd2+ during the hydrolysis of the phosphorothioate substrate in human plasma. In addition, the mechanism is postulated, by which a single internucleotide phosphorothioate bond of the SP-configuration at the 3′-end of unmodified phosphodiesters (PO-oligos), or their phosporothioate analogs (PS-oligos) protects these compounds against degradation in blood.

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Acknowledgments

The authors wish to thank Krzysztof Domanski for synthesis of the PS oligonucleotides 2 and 3. This study was supported by 502-17-692 project from Medical University of Łódź (to M.W.).

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Authors and Affiliations

  1. Department of Structural Biology, Medical University of Łódź, 7/9 Żeligowskiego St, 90-752, Lodz, Poland

    Marzena Wojcik

  2. Department of Bioorganic Chemistry, Center of Molecular and Macromolecular Studies, Polish Academy of Sciences, 112 Sienkiewicza St, 90-363, Lodz, Poland

    Wojciech J. Stec

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  1. Marzena Wojcik
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  2. Wojciech J. Stec
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Correspondence to Marzena Wojcik.

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The experimental work was carried out at the Department of Bioorganic Chemistry, Centre of Molecular and Macromolecular Studies of Polish Academy of Sciences in Lódź.

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Wojcik, M., Stec, W.J. The effect of divalent cations on the catalytic activity of the human plasma 3′-exonuclease. Biometals 23, 1113–1121 (2010). https://doi.org/10.1007/s10534-010-9358-5

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  • DOI: https://doi.org/10.1007/s10534-010-9358-5

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