Coordination compounds of copper have been invoked as major actors in processes involving the reduction of molecular oxygen, mostly with the generation of radical species the assignment for which has, so far, not been fully addressed. In the present work, we have carried out studies in solution and on surfaces to gain insights into the nature of the radical oxygen species (ROS) generated by a copper(II) coordination compound containing a thioether clip-phen derivative, 1,3-bis(1,10-phenanthrolin-2-yloxy)-N-(4-(methylthio)benzylidene)propan-2-amine (2CP-Bz-SMe), enabling its adsorption/immobilization to gold surfaces. Whereas surface plasmon resonance (SPR) and electrochemistry of the adsorbed complex indicated the formation of a dimeric Cu(I) intermediate containing molecular oxygen as a bridging ligand, scanning electrochemical microscopy (SECM) and nuclease assays pointed to the generation of a ROS species. Electron paramagnetic resonance (EPR) data reinforced such conclusions, indicating that radical production was dependent on the amount of oxygen and H2 O2 , thus pointing to a mechanism involving a Fenton-like reaction that results in the production of OH(.) .

Ruthenium polypyridine complexes have shown promise as agents for photodynamic therapy (PDT) and tools for molecular biology (chromophore-assisted light inactivation). To accomplish these tasks, it is important to have at least target selectivity and great reactive oxygen species (ROS) photogeneration: two properties that are not easily found in the same molecule. To prepare such new agents, we synthesized two new ruthenium complexes that combine an efficient DNA binding moiety (dppz ligand) together with naphthyl-modified (1) and anthracenyl-modified (2) bipyridine as a strong ROS generator bound to a ruthenium complex. The compounds were fully characterized and their photophysical and photochemical properties investigated. Compound 2 showed one of the highest quantum yields for singlet oxygen production ever reported (Φ= 0.96), along with very high DNA binding (log K = 6.78). Such photochemical behavior could be ascribed to the lower triplet state involving the anthracenyl-modified bipyridine, which is associated with easier oxygen quenching. In addition, the compounds exhibited moderate selectivity toward G-quadruplex DNA and binding to the minor groove of DNA, most likely driven by the pendant ligands. Interestingly, they also showed DNA photocleavage activity even upon exposure to a yellow light-emitting diode (LED). Regarding their biological activity, the compounds exhibited an exciting antibacterial action, particularly against Gram-positive bacteria, which was enhanced upon blue LED irradiation. Altogether, these results showed that our strategy succeeded in producing light-triggered DNA binding agents with pharmacological and biotechnological potential.

O composto 5-(4-pyridinyl)-1,3,4-oxadiazole-2-thiol (Hpyt) adsorve espontaneamente sobre ouro formando SAMs ("Self-Assembled Monolayers") que, de acordo com os resultados eletroquímicos e de STM ("Scanning Tunneling Microscopy"), contêm poros através dos quais as moléculas dos complexos [Fe(CN) 6 ] 4-, a dependência da corrente faradáica com o pH da solução eletrolítica permitiu o cálculo do pKa da molécula de Hpyt adsorvida sobre ouro (4,2). Os parâmetros termodinâmicos, H ads and G ads , para o processo de adsorção desta molécula foram estimados em -20,01 e -39,39 kJ mol -1 , respectivamente, utilizando-se o modelo de Langmuir. O processo redox da metaloproteína citocromo c foi estudado utilizando-se a SAM de Hpyt. A constante de velocidade heterogênea de transferência de elétrons foi calculada em 2,29 10 -3 cm s -1 .5-(4-pyridinyl)-1,3,4-oxadiazole-2-thiol (Hpyt) spontaneously adsorbs on gold forming SAMs (self-assembled monolayers) that, based on STM (Scanning Tunneling Microscopy) and electrochemical data, contain pinholes through which [Fe(CN) 6 ] 4-and [Ru(NH 3 ) 6 ] 3+ probe molecules access the underlying gold electrode. For the former molecule, the dependence of the faradaic current on the electrolyte solution pH value allowed the evaluation of the surface pKa as 4.2. The thermodynamic parameters H ads and G ads for the Hpyt adsorption process could be described by the Langmuir model and were calculated as -20.01 and -39.39 kJ mol -1 , respectively. Electrodic redox reaction of cytochrome c metalloprotein was accessed by using the Hpyt SAM with a heterogeneous electron transfer rate constant of 2.29 10 -3 cm s -1 .

The self-assembled monolayers (SAMs) derived from thionicotinamide (TNA), thioisonicotinamide (iTNA), and 5-(4-pyridyl)-1,3,4-oxadiazole-2-thiol (Hpyt) on gold have been characterized via surface-enhanced Raman scattering (SERS) as a function of pH and applied potential. Density functional theory calculations performed on the molecule/metal interaction model reinforced the vibrational assignments of the SERS spectra. Despite the structural similarity, these compounds presented different behaviors depending on the pH and applied potential with the iTNA SAM being the most affected. Upon adsorption and at pH 6, Hpyt SAM is not protonated while TNA and iTNA SAMs are partially and completely protonated, respectively. The results presented herein, besides being helpful for the understanding of the formation of the SAMs, can shed light on the understanding of the different responses observed for the cytochrome c metalloprotein by using the SAMs derived from these molecules.

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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 (+19mV), the binding to ligands that switch off the kinase activity induces a negative shift, i. e. E=-51, -57 and -156mV for O, imidazole and CN, respectively. Upon binding to CO, which does not affect the kinase active, E was observed at +21mV. 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.

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