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Fluorescence Labelling of DNA by Carboxylic Polypyridyl-Ru Complexes Containing bpy and DIP Ligands: A Study Revisited

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Abstract

The coordination complexes (DIP)2Ru(CH3bpyCOOH) and (DIP)2Ru(COOHbpyCOOH), where DIP and bpy are diphenylphenanthroline and bispyridine, have been recently proposed as fluorescent markers of nuclear DNA (Musatkina et al., J. Inorg. Biochem. 101:1086–1089, 2007), but no DNA binding investigation and no quantitative fluorescence evaluations had been done. Both complexes, as well as the smaller ones with bpy’s in place of DIP’s, have been investigated here by spectroscopic DNA titrations (UV–vis absorption, fluorescence, circular dichroism) and by in vitro cellular studies (flow cytometry and fluorescence imaging). Contrary to previous reports, neither the carboxylic function nor the more extended DIP ligand ensures any appreciable binding to DNA. This is clearly illustrated by the appearance of an isosbestic point of a second kind and by the proportionality of the fluorescence maximum intensity to the absorbance at the excitation wavelength. Above all, the lack of enhanced fluorescence in the presence of DNA definitively rules out the use of such complexes as DNA markers. Moreover, there is no detectable nuclear uptake. However, the fluorescent complexes with the DIP ligands, especially (DIP)2Ru(CH3bpyCOOH), are massively incorporated into the cytoplasm while preserving cell integrity, which could suggest other types of biological application.

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Acknowledgments

We are thankful to Dr. C. Cordier for having aroused our interest in these complexes. We are also very grateful to Dr. C. Cordier and Dr. H. Amouri for having provided us with the ruthenium complexes that had been synthesized by H. Amouri for their previous investigations. We thank Prof. J. Bolard for his advice on the spectroscopic approaches and Prof. J. Aubard for his encouragement. We are deeply grateful to Dr. J. Lomas for continuous support and fruitful discussions throughout this work.

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  1. Laboratoire Acides Nucléiques et Biophotonique, CNRS FRE 3207, Case courier 138, Université Pierre et Marie Curie, 4 place Jussieu, 75252, Paris Cedex, France

    Michèle Lamoureux & Olivier Seksek

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  1. Michèle Lamoureux
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  2. Olivier Seksek
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Correspondence to Michèle Lamoureux.

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Lamoureux, M., Seksek, O. Fluorescence Labelling of DNA by Carboxylic Polypyridyl-Ru Complexes Containing bpy and DIP Ligands: A Study Revisited. J Fluoresc 20, 631–643 (2010). https://doi.org/10.1007/s10895-009-0592-6

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