DNA affinity binding studies using a fluorescent dye displacement technique: the dichotomy of the binding site
Spillane, Caitriona B., Smith, Jayden A,, Morgan, Joy L., and Keene, F. Richard (2007) DNA affinity binding studies using a fluorescent dye displacement technique: the dichotomy of the binding site. Journal of Biological Inorganic Chemistry, 12 (6). pp. 819-824.
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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.
|Item Type:||Article (Refereed Research - C1)|
|Keywords:||bulge DNA; fluorescence assay; binding site; dinuclear ruthenium; binding selectivity; groove binding|
|Date Deposited:||08 May 2009 04:02|
|FoR Codes:||03 CHEMICAL SCIENCES > 0302 Inorganic Chemistry > 030201 Bioinorganic Chemistry @ 60%
03 CHEMICAL SCIENCES > 0302 Inorganic Chemistry > 030207 Transition Metal Chemistry @ 40%
|SEO Codes:||97 EXPANDING KNOWLEDGE > 970103 Expanding Knowledge in the Chemical Sciences @ 100%|
|Citation Count from Web of Science||