NMR analysis of duplex d(CGCGATCGCG)2 modified by Λ- and Δ-[Ru(bpy)2(m-GHK)]Cl2 and DNA photocleavage study
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- Myari, A., Hadjiliadis, N., Garoufis, A. et al. J Biol Inorg Chem (2007) 12: 279. doi:10.1007/s00775-006-0184-8
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The interaction of the diastereomeric complexes Λ-[Ru(bpy)2(m-GHK)]Cl2 and Δ-[Ru(bpy)2(m-GHK)]Cl2 (bpy is 2,2′-bipyridine, GHK is glycine–l-histidine–l-lysine) with the deoxynucleotide duplex d(5′-CGCGATCGCG)2 was studied by means of 1H NMR spectroscopy. At a Δ-isomer to DNA ratio of 1:1, significant shifts for the metal complex are observed, whereas there is negligible effect on the oligonucleotide protons and only one intermolecular nuclear Overhauser effect (NOE) is present at the 2D nuclear Overhauser enhancement spectroscopy spectrum. The 1Η NMR spectrum at ratio 2:1 is characterized by a slight shift for the Δ-isomer’s bpy aromatic protons as well as significant shifts for the decanucleotide G4 H1′ and Η2″, A5 H2, G10 H1′, T6 NH and G2 NH protons. Furthermore, at ratio 2:1, 11 intermolecular NOEs are observed. The majority of the NOEs involve the sugar Η2′ and Η2″ protons sited in the major groove of the decanucleotide. Increasing the Δ-isomer to d(CGCGATCGCG)2 ratio to 5:1 results in noteworthy spectral changes. The Δ-isomer’s proton shifts are reduced, whereas significant shifts are observed for the decanucleotide protons, especially the sugar protons, as well as for the exchangeable protons. Interaction is characterized by the presence of only one intermolecular NOE. Furthermore, there is significant broadening of the imino proton signals as the ratio of the Δ-isomer to DΝΑ increases, which is attributed to the opening of the two strands of the duplex. The Λ-isomer, on the other hand, approaches the minor groove of the oligonucleotide and interacts only weakly, possibly by electrostatic interactions. Photocleavage studies were also conducted with the plasmid pUC19 and a 158-bp restriction fragment, showing that both diastereomers cleave DNA with similar efficiency, attacking mainly the guanines of the sequence probably by generating active oxygen species.