The heme-based oxygen sensor Rhizobium etli FixL: influence of auxiliary ligands on heme redox potential and implications on the enzyme activity

Conformational changes associated to sensing mechanisms of heme-based protein sensors are a key molecular event that seems to modulate not only the protein activity but also the potential of the Fe redox couple of the heme domain. In this work, midpoint potentials (E) assigned to the Fe redox couple of the heme domain of FixL from Rhizobium etli (ReFixL) in the unliganded and liganded states were determined by spectroelectrochemistry in the presence of inorganic mediators. In comparison to the unliganded ReFixL protein (+ 19 mV), the binding to ligands that switch off the kinase activity induces a negative shift, i. e. E = − 51, − 57 and − 156 mV for O, imidazole and CN, respectively. Upon binding to CO, which does not affect the kinase active, E was observed at + 21 mV. The potential values observed for Fe of the heme domain of ReFixL upon binding to CO and O do not follow the expected trend based on thermodynamics, assuming that positive potential shift would be expected for ligands that bind to and therefore stabilize the Fe state. Our results suggest that the conformational changes that switch off kinase activity upon O binding have knock-on effects to the local environment of the heme, such as solvent rearrangement, destabilize the Fe state and counterbalances the Fe-stabilizing influence of the O ligand

Synergy of DNA intercalation and catalytic activity of a copper complex towards improved polymerase inhibition and cancer cell cytotoxicity

Improving the binding of metal complexes to DNA to boost cancer cell cytotoxicity requires fine tuning of their structural and chemical properties. Copper has been used as a metal center in compounds containing intercalating ligands due to its ability to catalytically generate reactive oxygen species (ROS), such as hydroxyl radicals (OH˙). We envision the synergy of DNA binding and ROS generation in proximity to target DNA as a powerful chemotherapy treatment. Here, we explore the use of [Cu(2CP-Bz-SMe)]2+(2CP-Bz-SMe = 1,3-bis(1,10-phenanthrolin-2-yloxy)-N-(4-(methylthio)benzylidene)propan-2-amine) for this purpose by characterizing its cytotoxicity, DNA binding, and ability to affect DNA replication through the polymerase chain reaction - PCR and nuclease assays. We determined the binding (Kb) and Stern-Volmer constants (KSV) for complex-DNA association of 5.8 ± 0.14 × 104and 1.64 (±0.08), respectively, through absorption titration and competitive fluorescence experiments. These values were superior to those of other Cu-complex intercalators. We hypothesize that the distorted trigonal bipyramidal geometry of [Cu(2CP-Bz-SMe)]2+allows the phenanthroline fragments to be better accommodated into the DNA double helix. Moreover, the aromaticity of these fragments increases the local hydrophobicity thus increasing the affinity for the hydrophobic domains of DNA. Nuclease assays in the presence of common reducing agents ascorbic acid, nicotinamide adenine dinucleotide, and glutathione showed the effective degradation of DNA due to thein situgeneration of OH˙. The [Cu(2CP-Bz-SMe)]2+complex showed cytotoxicity against the following human cancer cells lines A549, MCF-7, MDA-MB-231 and MG-63 with half maximal inhibitory concentration (IC50) values of 4.62 ± 0.48, 5.20 ± 0.76, 5.70 ± 0.42 and 2.88 ± 0.66 μM, respectively. These low values of IC50, which are promising if compared to that of cisplatin, are ascribed to the synergistic effect of ROS generation with the intercalation ability into the DNA minor grooves and blocking DNA replication. This study introduces new principles for synergizing the chemical and structural properties of intercalation compounds for improved drug-DNA interactions targeting cancer.Fil: Romo, Adolfo I. B.. University of Illinois. Urbana - Champaign; Estados Unidos. Universidade Federal do Ceara; BrasilFil: Carepo, Marta P.. Universidade Nova de Lisboa; PortugalFil: Levin, Pedro. Universidad de Santiago de Chile; ChileFil: Nascimento, Otaciro R.. Universidade Federal do São Carlos; BrasilFil: Díaz, Daniel E.. Universidad de Santiago de Chile; ChileFil: Rodriguez Lopez, Joaquin. University of Illinois. Urbana - Champaign; Estados UnidosFil: Leon, Ignacio Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Química Inorgánica "Dr. Pedro J. Aymonino". Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Química Inorgánica "Dr. Pedro J. Aymonino"; ArgentinaFil: Bezerra, Lucas F.. Universidade Federal do Ceara; BrasilFil: Lemus, Luis A.. Universidad de Santiago de Chile; ChileFil: Diógenes, Izaura C. N.. Universidade Federal do Ceara; Brasi

Asymmetric heterobimetallic mixed-valence complex trans-[(S'O IND.3')Co(cyclam)(NCS)Ru'(N'H IND.3') IND.4'(NCS)](B'F IND.4'): synthesis and characterization

[(SO3)Co(cyclam)(NCS)] and [(SO3)Co(cyclam)-NCS-Ru(NH3)4(NCS)](BF4) complexes were synthesized and characterized by means of X-ray diffraction, electrochemistry, elemental analysis, and spectroscopic techniques. Crystallographic and FTIR data indicated NCS- ligand is coordinated to Co through the nitrogen atom in the monomer species. Electrochemistry and FTIR data of the material isolated after reductive electrolysis of [(SO3)Co(cyclam)(NCS)] hint that NCS- and SO32- are released thus forming [Co(cyclam)(L)2]2+, where L is solvent molecules. The formation of the heterobimetallic mixed-valence complex induced a thermodynamic stabilization of Co and Ru metal atoms in the oxidized and reduced states, respectively. According to the Robin and Day classification, a Class II system with a comproportionation constant of 5.78 × 106 is suggested for the mixed-valence complex based on the electrochemical and UV-Vis-NIR results.CNPq (303530/2008-1)CAPESFAPESPFUNCA

On the correlation between electronic intramolecular delocalization and Au-S bonding strength of ruthenium tetraammine SAMs

Trans-[Ru(L)(NH3)4(L’)](PF6)n type complexes, where L = 4-cyanopyridine (CNpy), NCS-, CN-, and L’ = CNpy, 1,4-dithiane (1,4-dt), 4-mercaptopyridine (pyS) and thionicotinamide (tna), were synthesized and characterized. SAMs on gold of the complexes containing sulfur were studied by reductive desorption and SERS spectroscopy. Depending on the nature of L’, the withdrawing capability of the CNpy ligand is strong enough to partially oxidize the ruthenium atom and, as a consequence, delocalize the s electronic density from the trans located ligand. The reductive desorption results showed that the stability of the SAMs is directly related to this effect

Water-mediated reduction of [Cu(dmp)2(CH3CN)]2+: Implications of the structure of a classical complex on its activity as an anticancer drug

The interplay between coordination geometry and reactivity in copper complexes has been widely studied for years. ForCuII, it is known that the reduction is favored when the geometry is closer to that of the CuI state, which is mainly tetrahedral.Conversely, the reduction of CuII complexes in the absence of exogenous reducers has been barely addressed. Herein, wereport on the ability of a classic CuII complex to be partially reduced in acetonitrile solutions containing water, in the absenceof external reducers. The role of the structure on the spontaneous reduction is presented by contrasting the geometricfeatures with a related complex, [Cu(phen)2(CH3CN)]2+, which is inert towards the redox process mediated by water. Theparticipation of water in the reduction of [Cu(dmp)2(CH3CN)]2+ is associated with the production of hydroxyl radical. This,prompted us to evaluate the use of [Cu(dmp)2(CH3CN)]2+ as an anticancer metallodrug, showing an activity stronger than[Cu(phen)2(CH3CN)]2+ on 2D and 3D models of bone, lung, and breast cancer cell lines. Furthermore, both complexes aremore active than cisplatin. The main mechanism of action is the intracellular ROS generation, with a higher production ofcytotoxic species by [Cu(dmp)2(CH3CN)]2+. Certainly, the performance of [Cu(dmp)2(CH3CN)]2+ as an anticancer agent and itsreactivity in solution phase are connected through the geometrical constraints imparted by the dmp ligands.Fil: Levin, Pedro. Universidad de Santiago de Chile; ChileFil: Ruiz, Maria Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Química Inorgánica "Dr. Pedro J. Aymonino". Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Química Inorgánica "Dr. Pedro J. Aymonino"; ArgentinaFil: Romo, Adolfo Ignacio Barros. Universidade Federal do Ceara; BrasilFil: Nascimento, Otaciro Rangel. Universidade de Sao Paulo; BrasilFil: Di Virgilio, Ana Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Química Inorgánica "Dr. Pedro J. Aymonino". Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Química Inorgánica "Dr. Pedro J. Aymonino"; ArgentinaFil: Oliver, Allen Grayson. University of Notre Dame; Estados UnidosFil: Ayala, Alejandro P.. Universidade Federal do Ceara; BrasilFil: Diógenes, Izaura C. N.. Universidade Federal do Ceara; BrasilFil: Leon, Ignacio Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Química Inorgánica "Dr. Pedro J. Aymonino". Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Química Inorgánica "Dr. Pedro J. Aymonino"; ArgentinaFil: Lemus, Luis. Universidad de Santiago de Chile; Chil

Cation‐dependent stabilization of electrogenerated naphthalene diimide dianions in porous polymer thin films and their application to electrical energy storage

Porous polymer networks (PPNs) are attractive materials for capacitive energy storage because they offer high surface areas for increased double-layer capacitance, open structures for rapid ion transport, and redox-active moieties that enable faradaic (pseudocapacitive) energy storage. Here we demonstrate a new attractive feature of PPNs--the ability of their reduced forms (radical anions and dianions) to interact with small radii cations through synergistic interactions arising from densely packed redox-active groups, only when prepared as thin films. When naphthalene diimides (NDIs) are incorporated into PPN films, the carbonyl groups of adjacent, electrochemically generated, NDI radical anions and dianions bind strongly to K(+), Li(+), and Mg(2+), shifting the formal potentials of NDI's second reduction by 120 and 460 mV for K(+) and Li(+)-based electrolytes, respectively. In the case of Mg(2+), NDI's two redox waves coalesce into a single two-electron process with shifts of 240 and 710 mV, for the first and second reductions, respectively, increasing the energy density by over 20 % without changing the polymer backbone. In contrast, the formal reduction potentials of NDI derivatives in solution are identical for each electrolyte, and this effect has not been reported for NDI previously. This study illustrates the profound influence of the solid-state structure of a polymer on its electrochemical response, which does not simply reflect the solution-phase redox behavior of its monomers.Porous polymer networks (PPNs) are attractive materials for capacitive energy storage because they offer high surface areas for increased double‐layer capacitance, open structures for rapid ion transport, and redox‐active moieties that enable faradaic (ps54451322513229FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO2013/25527‐1This research was supported by an NSF GRFP (DGE‐1144153) award to C.R.D. W.R.D. acknowledges support from the Alfred P. Sloan and Camille and Henry Dreyfus Foundations. This work was supported in part (K.H.B., H.D.A.) through grant DE‐FG02‐87ER45298, by

Ascorbyl and hydroxyl radical generation mediated by a copper complex adsorbed on gold

This work presents the results obtained for a thioether derivative of bipyridine, (E,Z)-1-(4'-methyl-[2,2'-bipyridine]-4-yl)-N-(4(methylthio)phenyl)methanimine (4-mbpy-Bz-SMe), and its copper complex [CuII(4-mbpy-Bz-SMe)2]2+. Electronic spectra acquired at 183 K of the cuprous complex [CuI(4-mbpy-Bz-SMe)2]+ generated in situ indicated the formation of the peroxodicopper compound {[CuII(4-mbpy-Bz-SMe)2]2(μ-O22-)}2+. A gold electrode modified with [CuII(4-mbpy-Bz-SMe)2]2+ (Au/[Cu]) was fully characterized by SERS spectroscopy, electrochemistry and impedance spectroscopy thus showing adsorption occurs through the sulfur atom of the 4-mbpy-Bz-SMe moieties. DNA cleavage assays showed the copper complex, in solution and adsorbed on gold, degrades DNA if reducing conditions are maintained, i.e. ascorbic acid (H2AA) in solution or applied potentials more negative than 0.12 V vs. Ag/AgCl (CuI form). The electron paramagnetic resonance (EPR) spectra obtained for the electrolyzed solution (Eapl = -0.2 V, no H2O2) and for the solution containing [CuII(4-mbpy-Bz-SMe)2]2+ and H2O2 showed hydroxyl radical, HO˙, generation had occurred. The cyclic voltammograms obtained with H2AA in solution at Au/[CuII(4-mbpy-Bz-SMe)2]2+ as the working electrode showed a one-electron reaction leading to the ascorbyl radical (HA˙), which was detected by EPR. The current assigned to the electrode oxidation of HA˙ to AA decreased with the addition of catalase, a scavenger of H2O2, meaning peroxide is involved in the mechanism