New insights in the DNA-[Cr(phen)2(dppz)]3+ binding and photocleavage properties by the complex with an intercalating ligand

• Autores: Toneatto, Judith; Boero, Rodolfo Alejandro; Lorenzatti, Guadalupe; Cabanillas, Ana Maria de Los A.; Arguello, Gerardo
• Año de publicación: 2010
• Idioma: inglés
• Tipo de recurso: artículo
• Materia: CHROMIUM COMPLEX; PHOTOSENSITIZATION; INTERCALATING LIGAND; DNA BINDING; DNA PHOTOCLEAVAGE; BACTERIAL TRANSFORMATION; BACTERIAL GROWTH PHOTOINHIBITION
• Institución: Consejo Nacional de Investigaciones Científicas y Técnicas

Due to the key role of DNA in cell life and pathological processes, the design of specific chemical nucleases, fic chemical nucleases, DNA probes and alkylating agents is an important research area for the development of new therapeutic agents and tools in Biochemistry. Hence, the interaction of small molecules with DNA has attracted in particular a great deal of attention. 27a great deal of attention. The aim of this study was to investigate the ability of [Cr(phen)2(dppz)]3+ to associate with DNA and to 282(dppz)]3+ to associate with DNA and to characterize it as photocleavage reagent for Photodynamic Therapy (PDT).Chromium(III) complex [Cr(phen)2(dppz)]3+, (dppz=dipyridophenazine, phen=1,10-phenanthroline), 302(dppz)]3+, (dppz=dipyridophenazine, phen=1,10-phenanthroline),where dppz is a planar bidentate ligand with an extended ¦Ð system, has been found to bind strongly 31¦Ð system, has been found to bind strongly to double strand oligonucleotides (ds-oligo) and plasmid DNA with intrinsic DNA binding constants, Kb, of 32ds-oligo) and plasmid DNA with intrinsic DNA binding constants, Kb, of (3.9¡À0.3)¡Á105 M−1 and (1.1¡À0.1)¡Á105 M−1, respectively. The binding properties to DNA were 335 M−1 and (1.1¡À0.1)¡Á105 M−1, respectively. The binding properties to DNA were investigated by UV¨Cvisible (UV¨CVis) absorption spectroscopy and electrophoretic studies. UV¨CVis absorption 34¨Cvisible (UV¨CVis) absorption spectroscopy and electrophoretic studies. UV¨CVis absorption data provide clearly that the chromium(III) complex interacts with DNA intercalatively. Competitive binding experiments show that the enhancement in the emission intensity of ethidium bromide (EthBr) in the presence of DNA was quenched by [Cr(phen)2(dppz)]3+, indicating that the Cr(III) complex displaces EthBr 372(dppz)]3+, indicating that the Cr(III) complex displaces EthBr from its binding site in plasmid DNA. Moreover, [Cr(phen)2(dppz)]3+, non-covalently bound to 382(dppz)]3+, non-covalently bound to DNA, promotes the photocleavage of plasmid DNA under 457 nm irradiation. We also found that the irradiated Cr(III)-plasmid DNA association is able to impair the transforming capacity of bacteria. These results provide evidence confirming the responsible and essential role of the excited state of [Cr(phen)2(dppz)]3+ for firming the responsible and essential role of the excited state of [Cr(phen)2(dppz)]3+ for damaging the DNA structure. The combination of DNA, [Cr(phen)2(dppz)]3+ and light, is necessary to induce damage. In addition, assays of the photosensitization of transformed bacterial suspensions suggest that Escherichia coli may be photoinactivated by irradiation in the presence of [Cr(phen)2(dppz)]3+. In sum, our may be photoinactivated by irradiation in the presence of [Cr(phen)2(dppz)]3+. In sum, our results allow us to postulate the [Cr(phen)2(dppz)]3+ complex as a very attractive candidate for DNA 2(dppz)]3+ complex as a very attractive candidate for DNA photocleavage with potential applications in Photodynamic Therapy (PDT).fic chemical nucleases, DNA probes and alkylating agents is an important research area for the development of new therapeutic agents and tools in Biochemistry. Hence, the interaction of small molecules with DNA has attracted in particular a great deal of attention. 27a great deal of attention. The aim of this study was to investigate the ability of [Cr(phen)2(dppz)]3+ to associate with DNA and to 282(dppz)]3+ to associate with DNA and to characterize it as photocleavage reagent for Photodynamic Therapy (PDT).Chromium(III) complex [Cr(phen)2(dppz)]3+, (dppz=dipyridophenazine, phen=1,10-phenanthroline), 302(dppz)]3+, (dppz=dipyridophenazine, phen=1,10-phenanthroline),where dppz is a planar bidentate ligand with an extended ¦Ð system, has been found to bind strongly 31¦Ð system, has been found to bind strongly to double strand oligonucleotides (ds-oligo) and plasmid DNA with intrinsic DNA binding constants, Kb, of 32ds-oligo) and plasmid DNA with intrinsic DNA binding constants, Kb, of (3.9¡À0.3)¡Á105 M−1 and (1.1¡À0.1)¡Á105 M−1, respectively. The binding properties to DNA were 335 M−1 and (1.1¡À0.1)¡Á105 M−1, respectively. The binding properties to DNA were investigated by UV¨Cvisible (UV¨CVis) absorption spectroscopy and electrophoretic studies. UV¨CVis absorption 34¨Cvisible (UV¨CVis) absorption spectroscopy and electrophoretic studies. UV¨CVis absorption data provide clearly that the chromium(III) complex interacts with DNA intercalatively. Competitive binding experiments show that the enhancement in the emission intensity of ethidium bromide (EthBr) in the presence of DNA was quenched by [Cr(phen)2(dppz)]3+, indicating that the Cr(III) complex displaces EthBr 372(dppz)]3+, indicating that the Cr(III) complex displaces EthBr from its binding site in plasmid DNA. Moreover, [Cr(phen)2(dppz)]3+, non-covalently bound to 382(dppz)]3+, non-covalently bound to DNA, promotes the photocleavage of plasmid DNA under 457 nm irradiation. We also found that the irradiated Cr(III)-plasmid DNA association is able to impair the transforming capacity of bacteria. These results provide evidence confirming the responsible and essential role of the excited state of [Cr(phen)2(dppz)]3+ for firming the responsible and essential role of the excited state of [Cr(phen)2(dppz)]3+ for damaging the DNA structure. The combination of DNA, [Cr(phen)2(dppz)]3+ and light, is necessary to induce damage. In addition, assays of the photosensitization of transformed bacterial suspensions suggest that Escherichia coli may be photoinactivated by irradiation in the presence of [Cr(phen)2(dppz)]3+. In sum, our may be photoinactivated by irradiation in the presence of [Cr(phen)2(dppz)]3+. In sum, our results allow us to postulate the [Cr(phen)2(dppz)]3+ complex as a very attractive candidate for DNA 2(dppz)]3+ complex as a very attractive candidate for DNA photocleavage with potential applications in Photodynamic Therapy (PDT).
Fil: Toneatto, Judith. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Boero, Rodolfo Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina
Fil: Lorenzatti, Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Cabanillas, Ana Maria de Los A.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Arguello, Gerardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina