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DNA affinity binding studies using a fluorescent dye displacement technique: the dichotomy of the binding site

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Abstract

We have observed a number of discrepancies and contradictions in the use of a fluorescent intercalator displacement assay in surveying the binding affinities of dinuclear polypyridyl ruthenium(II) complexes with DNA. By a modification of the assay using the fluorescent minor-groove binder 4′,6-diamidino-2-phenylindole, rather than intercalating dyes (ethidium bromide or thiazole orange), results were obtained for all complexes studied which were consistent with relative affinities and stereoselectivities observed with other techniques, including NMR, affinity chromatography and equilibrium dialysis. It is believed that the difference in binding mode between the minor groove-binding Ru(II) complexes and the intercalating fluorescent dyes they are displacing may contribute to these discrepancies.

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Acknowledgement

We gratefully acknowledge the financial support of the Australian Research Council.

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Authors and Affiliations

  1. School of Pharmacy and Molecular Sciences, James Cook University, Townsville, QLD, 4811, Australia

    Caitriona B. Spillane, Jayden A. Smith, Joy L. Morgan & F. Richard Keene

Authors
  1. Caitriona B. Spillane
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  2. Jayden A. Smith
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  3. Joy L. Morgan
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  4. F. Richard Keene
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Corresponding author

Correspondence to F. Richard Keene.

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Spillane, C.B., Smith, J.A., Morgan, J.L. et al. DNA affinity binding studies using a fluorescent dye displacement technique: the dichotomy of the binding site. J Biol Inorg Chem 12, 819–824 (2007). https://doi.org/10.1007/s00775-007-0235-9

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  • DOI: https://doi.org/10.1007/s00775-007-0235-9

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