DNA affinity binding studies using a fluorescent dye displacement technique: the dichotomy of the binding site |
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Authors: | Caitriona B Spillane Jayden A Smith Joy L Morgan F Richard Keene |
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Institution: | (1) School of Pharmacy and Molecular Sciences, James Cook University, Townsville, QLD, 4811, Australia |
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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.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. |
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Keywords: | Bulge DNA Dinuclear ruthenium Fluorescence assay 4′ 6-Diamidino-2-phenylindole Binding selectivity |
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