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Binding and transfer of an oligodeoxynucleotide containing the translation initiation site of the BCL2 mRNA into K562 cells

  • Cell Molecular Biology
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Abstract

A study was conducted of the interaction of an octadecameric oligodeoxynucleotide (dON) containing the BCL2 mRNA translation start with K562 cells. Both in solution and upon lipofection, the binding of dON used at a low concentration (≤30 nM) at 37°C involved two steps: saturating surface binding with the cell membrane and internalization. Three phases were revealed in the dynamics of internalization: the extent and rate of internalization increased during the first hour of incubation; decreased during the second hour; and then increased again, which was assumed to reflect the priming of new dON-binding sites. The binding constant and the number of binding sites were estimated at 10°C (the conditions preventing internalization) by consecutive dissociation of dON-cell complexes formed in 1 h at 37°C. Incubation of dON with cells led to the priming of new high-affinity binding sites and an increase of the binding constant to a level characteristic of high-affinity ligand-receptor interactions (109 M−1). High-affinity receptor-mediated binding preceded internalization of dON. Lipofection increased the binding constant and the number of binding sites severalfold but had virtually no effect on the temporal pattern and the extent of dON internalization.

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Authors and Affiliations

  1. Hematology Research Center, Russian Academy of Medical Sciences, Moscow, 125167, Russia

    A. M. Timofeev, T. V. Borovkova, N. M. Nydenova, T. V. Akhlynina, D. A. Shmarov & N. I. Grineva

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  1. A. M. Timofeev
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  2. T. V. Borovkova
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  3. N. M. Nydenova
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  4. T. V. Akhlynina
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  5. D. A. Shmarov
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  6. N. I. Grineva
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__________

Translated from Molekulyarnaya Biologiya, Vol. 39, No. 2, 2005, pp. 235–244.

Original Russian Text Copyright © 2005 by Timofeev, Borovkova, Nydenova, Akhlynina, Shmarov, Grineva.

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Timofeev, A.M., Borovkova, T.V., Nydenova, N.M. et al. Binding and transfer of an oligodeoxynucleotide containing the translation initiation site of the BCL2 mRNA into K562 cells. Mol Biol 39, 210–217 (2005). https://doi.org/10.1007/s11008-005-0031-y

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  • DOI: https://doi.org/10.1007/s11008-005-0031-y

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