title: Binding of oxindole-Schiff base copper(II) complexes to DNA and its modulation by the ligand.
abstract: Previous studies on copper(II) complexes with oxindole-Schiff base ligands have shown their potential antitumor activity towards different cells, inducing apoptosis through a preferential attack to DNA and/or mitochondria. Herein, we better characterize the interactions between some of these copper(II) complexes and DNA. Investigations on its binding ability to DNA were carried out by fluorescence measurements in competitive experiments with ethidium bromide, using plasmidial or calf-thymus DNA. These results indicated an efficient binding process similar to that observed with copper(II)-phenanthroline species, [Cu(o-phen)(2)](2+), with binding constants in the range 3 to 9×10(2) M(-1). DNA cleavage experiments in the presence and absence of distamycin, a recognized binder of DNA, indicated that this binding probably occurs at major or minor groove, leading to double-strand DNA cleavage, and being modulated by the imine ligand. Corroborating these data, discrete changes in EPR spectra of the studied complexes were observed in the presence of DNA, while more remarkable changes were observed in the presence of nucleotides (AMP, GMP, CMP or UMP). Additional evidence for preferential coordination of the copper centers to the bases guanine or cytosine was obtained from titrations of these complexes with each nucleotide, monitored by absorption spectral changes. Therefore, the obtained data point out to their action as groove binders to DNA bases, rather than as intercalators or covalent cross-linkers. Further investigations by SDS PAGE using (32)P-ATP or (32)P-oligonucleotides attested that no hydrolysis of phosphate linkage in DNA or RNA occurs, in the presence of such complexes, confirming their main oxidative mechanism of action.
authors.affiliation: Departamento de Química Fundamental, Instituto de Química, Universidade de São Paulo, São Paulo, Brazil.
authors.forename: Vivian Chagas
authors.initials: VC
authors.lastname: da Silveira
authors.forename: Henri
authors.initials: H
authors.lastname: Benezra
authors.forename: Juliana Silva
authors.initials: JS
authors.lastname: Luz
authors.forename: Raphaela Castro
authors.initials: RC
authors.lastname: Georg
authors.forename: Carla Columbano
authors.initials: CC
authors.lastname: Oliveira
authors.forename: Ana Maria da Costa
authors.initials: AM
authors.lastname: Ferreira
mesh_terms: D000465:Algorithms; D001667:Binding, Competitive; D002942:Circular Dichroism; D056831:Coordination Complexes / Q000737:chemistry*; D003300:Copper; D004247:DNA / Q000737:chemistry*; D053837:DNA Cleavage; D004270:DNA, Circular / Q000737:chemistry*; D004214:Distamycins / Q000737:chemistry; D004578:Electron Spin Resonance Spectroscopy; D004996:Ethidium / Q000737:chemistry; D007211:Indoles / Q000737:chemistry*; D007364:Intercalating Agents / Q000737:chemistry; D000078183:Oxindoles; D012545:Schiff Bases / Q000737:chemistry; D013050:Spectrometry, Fluorescence
chemical_list: D056831:Coordination Complexes; D004270:DNA, Circular; D004214:Distamycins; D007211:Indoles; D007364:Intercalating Agents; D000078183:Oxindoles; D012545:Schiff Bases; C022960:2-oxindole; D003300:Copper; C030000:stallimycin; D004247:DNA; C427203:calf thymus DNA; D004996:Ethidium