Deciphering the Impact of Nucleosides and Nucleotides on Copper Ion and Dopamine Coordination Dynamics.

The mode of coordination of copper(II) ions with dopamine (DA, L) in the binary, as well as ternary systems with Ado, AMP, ADP, and ATP (L') as second ligands, was studied with the use of experimental-potentiometric and spectroscopic (VIS, EPR, NMR, IR)-methods and computational-molecular modeling and DFT-studies. In the Cu(II)/DA system, depending on the pH value, the active centers of the ligand involved in the coordination with copper(II) ions changed from nitrogen and oxygen atoms (CuH(DA)3+, Cu(DA)2+), via nitrogen atoms (CuH2(DA)24+), to oxygen atoms at strongly alkaline pH (Cu(DA)22+). The introduction of L' into this system changed the mode of interaction of dopamine from oxygen atoms to the nitrogen atom in the hydroxocomplexes formed at high pH values. In the ternary systems, the ML'-L (non-covalent interaction) and ML'HxL, ML'L, and ML'L(OH)x species were found. In the Cu(II)/DA/AMP or ATP systems, mixed forms were formed up to a pH of around 9.0; above this pH, only Cu(II)/DA complexes occurred. In contrast to systems with AMP and ATP, ternary species with Ado and ADP occurred in the whole pH range at a high concentration, and moreover, binary complexes of Cu(II) ions with dopamine did not form in the detectable concentration.

Intermolecular interactions of tetracyanoethylene (TCNE) and fumaronitrile (FN) with minor amines: A combined UV-Vis and EPR study.

In this paper, the nature of interactions between two cyanocarbons-tetracyanoethylene (TCNE) and fumaronitrile (FN)-and a series of four secondary amines possessing a general formula C4HxN (x = 5-11) is thoroughly scrutinized. For all of the TCNE-amine pairs, tricyanovinylation (TCV) reaction is observed; however, only for pyrrole, it is accompanied with a visible charge-transfer (CT) complex formation-no such chemical individuals, characteristic for TCNE, have been noticed for aliphatic and alicyclic amines. On the contrary, FN forms such complexes with all the amines studied. Interestingly, a rather unexpected reaction of FN with alicyclic amines has been observed. The recorded electron paramagnetic resonance (EPR) spectra indicate the presence of both TCNE●- and FN●- radicals in the analyzed samples, assigned to a complete charge (electron) transfer process within the CT complexes, whose efficiency can be additionally enhanced by photoirradiation. The origination of the former radical, whose presence is observed also in the TCNE-diethylamine mixture, is as well proposed to result indirectly from the TCV reaction, occurring for this system. Finally, the superhyperfine structure of EPR spectra, indicating the existence of some secondary interactions of the radicals with surrounding compounds, is discussed. Formation of CT complexes and tricyanovinylates has been investigated and characterized with UV-Vis spectroscopy, while the presence of (cyano)radicals in the analyzed mixtures has been evidenced by (photoinduced) EPR measurements. Interpretation of the experimental results is also supplemented with computer simulations including density functional theory calculations.

Analysis of NH3 -TPD Profiles for CuSSZ-13 SCR Catalyst of Controlled Al Distribution - Complexity Resolved by First Principles Thermodynamics of NH3 Desorption, IR and EPR Insight into Cu Speciation*.

NH3 temperature-programmed desorption (NH3 -TPD) is frequently used for probing the nature of the active sites in CuSSZ-13 zeolite for selective catalytic reduction (SCR) of NOx . Herein, we propose an interpretation of NH3 -TPD results, which takes into account the temperature-induced dynamics of NH3 interaction with the active centers. It is based on a comprehensive DFT/GGA+D and first-principles thermodynamic (FPT) modeling of NH3 adsorption on single Cu2+ , Cu+ , [CuOH]+ centers, dimeric [Cu-O-Cu]2+ , [Cu-O2 2- -Cu]2 species, segregated CuO nanocrystals and Brønsted acid sites (BAS). Theoretical TPD profiles are compared with the experimental data measured for samples of various Si/Al ratios and distribution of Al within the zeolite framework. Copper reduction, its relocation, followed by the intrazeolite olation/oxolation processes, which are concomitant with NH3 desorption, were revealed by electron paramagnetic resonance (EPR) and IR. DFT/FPT results show that the peaks in the desorption profiles cannot be assigned univocally to the particular Cu and BAS centers, since the observed low-, medium- and high-temperature desorption bands have contributions coming from several species, which dynamically change their speciation and redox states during NH3 -TPD experiment. Thus, a rigorous interpretation of the NH3 -TPD profiles of CuSSZ-13 in terms of the strength and concentration of the active centers of a particular type is problematic. Nonetheless, useful connections for molecular interpretation of TPD profiles can be established between the individual component peaks and the corresponding ensembles of the adsorption centers.

The Cytotoxic Effect of Copper (II) Complexes with Halogenated 1,3-Disubstituted Arylthioureas on Cancer and Bacterial Cells.

A series of eight copper (II) complexes with 3-(4-chloro-3-nitrophenyl)thiourea were designed and synthesized. The cytotoxic activity of all compounds was assessed in three human cancer cell lines (SW480, SW620, PC3) and human normal keratinocytes (HaCaT). The complexes 1, 3, 5, 7 and 8 were cytotoxic to the studied tumor cells in the low micromolar range, without affecting the normal cells. The complexes 1, 3, 7 and 8 induced lactate dehydrogenase (LDH) release in all cancer cell lines, but not in the HaCaT cells. They provoked early apoptosis in pathological cells, especially in SW480 and PC3 cells. The ability of compounds 1, 3, 7 and 8 to diminish interleukin-6 (IL-6) concentration in a cell was established. For the first time, the influence of the most promising Cu (II) complexes on intensities of detoxifying and reactive oxygen species (ROS) scavenging the enzymes of tumor cells was studied. The cytotoxic effect of all copper (II) conjugates against standard and hospital bacterial strains was also proved.

Cu(II) Complexes with FomA Protein Fragments of Fusobacterium Nucleatum Increase Oxidative Stress and Malondialdehyde Level.

An explanation of carcinogenesis processes may certainly contribute to the prevention and development of novel methods for cancer treatment. In this paper, we considered the probable relationship between the presence of Fusobacterium nucleatum in the colon and its possible influence on the development of colorectal cancer. For this purpose, intracellular and/or extracellular generation of reactive oxygen species (ROS) by mouse colon carcinoma cells (CT26) was stimulated by two fragments of FomA adhesin from F. nucleatum and their complexes with copper(II): Cu(II)-Ac-KGHGNG-NH2 (1Cu) and Cu(II)-Ac-PTVHNE-NH2 (2Cu). Incubation of the cells with copper complexes was followed with ICP-MS technique. The overall generation of ROS was shown by means of fluorescence spectroscopy with two proper probes, whereas identification of ROS was achieved by the spin trapping technique and electron paramagnetic resonance measurements. As a result, an abundant production of the hydroxyl radicals, both inside and outside the cells, was observed upon the stimulation of the CT26 cells with the copper complexes. Clearly both compounds induced strong oxidation stress which triggered a radicals' cascade that finally resulted in the pronounced lipid peroxidation. The latter was evidenced with the measured level of malondialdehyde, a biomarker of the peroxidation process. By applying N-acetylcysteine antioxidant to the studied system, the free radical mechanism of the lipid peroxidation process was confirmed. Hypothetically this mechanism can lead to colon cell damage and further cancerogenesis processes.

Paramagnetic dioxovanadium(IV) molecules inside the channels of zeolite BEA--EPR screening of VO2 reactivity toward small gas-phase molecules.

Interaction of small gas-phase molecules (NO, N2O, O2, CO) with VO2 radicals inside the channels of a dealuminated SiBEA zeolite was investigated by means of electron paramagnetic resonance (EPR), infrared (IR), and mass (QMS) spectroscopies to provide direct insights into the chemistry of a unique paramagnetic state of vanadium - VO2 molecules. A facile way of forming VO2 inside the channels of SiBEA via thermal reduction of VO2(+) precursor cations was shown. Dioxovanadium(IV) was identified based on its unusual EPR signal which, as compared with the typical monooxovanadium(IV) (VO(2+) cation), is featured by rhombic symmetry and a positive Aiso value leading to a hyperfine splitting as large as 32 mT. VO2 molecules exhibit reducing properties transforming N2O and O2 into vanadium intrachannel cage adducts comprising of reactive oxygen species (O(-) and O2(-), respectively). Interaction with CO led to its oxidation to CO2, while paramagnetic NO acted as a scavenger for VO2 radicals producing diamagnetic adducts. The observed reactivity was rationalized in terms of spin-pairing, electron transfer, and oxygen transfer processes. As a result new chemical pathways of vanadium reactivity were demonstrated which were not observed so far either in the homogeneous molecular systems or supported vanadium materials.

Intimate binding mechanism and structure of trigonal nickel(I) monocarbonyl adducts in ZSM-5 zeolite-spectroscopic continuous wave EPR, HYSCORE, and IR studies refined with DFT quantification of disentangled electron and spin density redistributions along σ and π channels.

Interaction of tetracoordinated nickel(I) centers generated inside the channels of ZSM-5 zeolite with carbon monoxide ((12,13)CO, pCO < 1 Torr) led to the formation of T-shaped, top-on monocarbonyl adducts with a unique trigonal nickel core, supported by two oxygen donor ligands. The mechanism of the formation of the {Ni(I)-CO}ZSM-5 species was accounted for by a quantitative molecular orbital correlation diagram of CO ligation. Detailed electronic and magnetic structure of this adduct was obtained from comprehensive DFT calculations, validated by quantitative reproduction of its continuous wave electron paramagnetic resonance (CW-EPR), hyperfine sublevel correlation (HYSCORE), and IR fingerprints, using relativistic Pauli and ZORA-SOMF/B3LYP methods. Molecular analysis of the stretching frequency, νCO = 2109 cm(-1), g and A((13)C) tensors (g(xx) = 2.018, g(yy) = 2.380, g(zz) = 2.436, A(xx) = +1.0 ± 0.3 MHz, A(yy) = -3.6 ± 0.9 MHz, A(zz) = -1.6 ± 0.3 MHz) and Q((27)Al) parameters (e(2)Qq/h = -13 MHz and η = 0.8) supported by quantum chemical modeling revealed that the Ni-CO bond results from the π overlap between the low-laying π(2p) CO states with the 3d(xz) and 3d(yz) orbitals, with a small σ contribution due to the overlap of σ(2p+2s) orbital and a protruding lobe of the in-plane 3d(xz) orbital. Two types of orbital channels (associated with the σ and π overlap) of the electron and spin density flows within the {Ni(I)-CO} unit were identified. A bathochromic shift of the νCO stretching vibration was accounted for by resolving quantitatively the separate contributions due to the σ donation and π back-donation, whereas the (13)C hyperfine coupling was rationalized by incongruent α and ß spin flows via the σ and π channels. As a result the very nature of the carbon-metal bond in the Ni(I)-CO adduct and the molecular backbone of the corresponding spectroscopic parameters were revealed with unprecedented accuracy.

Spectroscopic IR, EPR, and operando DRIFT insights into surface reaction pathways of selective reduction of NO by propene over the Co-BEA zeolite.

Interaction of a Co-BEA catalyst with individual components (NO, C(3)H(6), CO, O(2)) and mixtures simulating the real feed of the selective catalytic reduction (SCR) of nitric oxide in static and pulse experiments at variable temperatures was investigated by means of IR, EPR, and operando DRIFT spectroscopy coupled with QMS/GC analysis of the products. Speciation of cobalt active sites into Co(II), mono- and polynuclear oxo-cobalt species as well as CoO clusters was quantified by IR using CO and NO as probe molecules. The key intermediates, by-products, and final products of the SCR reaction were identified and their spectroscopic signatures ascertained. Based on the spectroscopic operando results, a concise mechanistic scheme of the selective catalytic reduction of nitric oxide by propene, triggered by a two-electron Co(II)/Co(0) redox couple, was developed. It consists of a complex network of the sequential/parallel selective reduction steps that are interlocked by the rival nonselective oxidation of the intermediates and their thermal decomposition. It has been shown that the SCR process is initiated by the chemoselective capture of NO from the reaction mixture by the cobalt active sites leading to the cobalt(II) dinitrosyls, which in the excess of oxygen are partially oxidized to surface nitrates and nitrites. N(2)O is produced by semi-decomposition of the dinitrosyl intermediates on the mononuclear centers, whereas NO(2)via NO oxidation on the polynuclear oxo-cobalt sites. Cyanide and isocyanate species, formed together with propene oxygenates in the course of the C=C bond scission, are the mechanistically pivotal reaction intermediates of C(3)H(6) interaction with the dinitrosyles and NO(3)(-)/NO(2)(-) surface species. Dinitrogen is produced by three main reaction routes involving oxidation of cyanides by NO/NO(2), reduction of dinitrosyls, nitrates, and nitrites by propene oxygenates (medium temperature range) or their reduction by carbon monoxide (high temperature range).

Unraveling the Origin of Enhanced Activity of the Nb2O5/H2O2 System in the Elimination of Ciprofloxacin: Insights into the Role of Reactive Oxygen Species in Interface Processes.

The overlooked role of reactive oxygen species (ROS), formed and stabilized on the surface of Nb2O5 after H2O2 treatment, was investigated in the adsorption and degradation of ciprofloxacin (CIP), a model antibiotic. The contribution of ROS to the elimination of CIP was assessed by using different niobia-based materials in which ROS were formed in situ or ex situ. The formation of ROS was confirmed by electron paramagnetic resonance (EPR) and Raman spectroscopy. The modification of the niobia surface charge by ROS was monitored with zeta potential measurements. The kinetics of CIP removal was followed by UV-vis spectroscopy, while identification of CIP degradation products and evaluation of their cytotoxicity were obtained with liquid chromatography-mass spectrometry (LC-MS) and microbiological studies, respectively. Superoxo and peroxo species were found to significantly improve the efficiency of CIP adsorption on Nb2O5 by modifying its surface charge. At the same time, it was found that improved removal of CIP in the dark and in the presence of H2O2 was mainly determined by the adsorption process. The enhanced adsorption was confirmed by infrared spectroscopy (IR), total organic carbon measurements (TOC), and elemental analysis. Efficient chemical degradation of adsorbed CIP was observed upon exposure of the Nb2O5/H2O2 system to UV light. Therefore, niobia is a promising inorganic adsorbent that exhibits enhanced sorption capacity toward CIP in the presence of H2O2 under dark conditions and can be easily regenerated in an environmentally benign way by irradiation with UV light.

Phosphate doping as a promising approach to improve reactivity of Nb2O5 in catalytic activation of hydrogen peroxide and removal of methylene blue via adsorption and oxidative degradation.

This study is devoted to the evaluation of the influence of phosphate dopants on the reactivity of Nb2O5-based nanomaterials in the combined catalytic activation of H2O2 and the elimination of methylene blue (MB) from an aqueous solution via adsorption and chemical degradation. For this purpose, several niobia-based catalysts doped with various amounts of phosphate were prepared by a facile hydrothermal method and subsequent calcination. Phosphate doping was shown to strongly enhance the ability of Nb2O5 to activate H2O2, as well as to adsorb and degrade MB. The most pronounced differences in the reactivity of the parent Nb2O5 and phosphate-doped samples were observed under strongly acidic conditions (pH ~ 2.4), at which the most active modified catalysts (Nb/P molar ratio = 5/1) was approximately 6 times more efficient in the removal of MB. The observed enhancement of reactivity was attributed to the increased generation of singlet oxygen 1O2, which was identified as the main oxidizing agent responsible for efficient degradation of MB. To our knowledge, it is the first report revealing that phosphate doping of Nb2O5 resulted in an improved activity of niobia in the adsorption and degradation of organic pollutants.

Relevance of the electron transfer pathway in photodynamic activity of Ru(II) polypyridyl complexes containing 4,7-diphenyl-1,10-phenanthroline ligands under normoxic and hypoxic conditions.

The purpose of this study was to investigate the correlation between the spectroscopic and photophysical properties of Ru(II) polypyridyl complexes and their photodynamic activity in vitro. A series of Ru(II) polypyridyl complexes with 4,7-diphenyl-1,10-phenanthroline (dip) and 2,3-bis(2-pyridyl)quinoxaline (dpq) and its derivatives were synthesized and characterized regarding their photophysical, biological, and photodynamic properties. The complexes were evaluated not only in the context of 1O2 generation but also regarding other types of reactive oxygen species (ROS) to assess the possibility of Ru(II) complexes to induce phototoxicity via various ROS using fluorescence and EPR spectroscopy. The compounds were found to be moderately cytotoxic with IC50 values ranging from 1 to 35 µM and retained their cytotoxic activity under hypoxic conditions. The unraveled phototoxic activity is based mainly on the generation of H2O2 and 1O2, highlighting the importance of electron-transfer processes in the observed photodynamic activity of Ru polypyridyl complexes. A combination of photodynamic activity with cytotoxicity under decreased dioxygen concentrations may help overcome the current photodynamic therapy (PDT) limitation. The findings highlight the need for broadening the scope of tested Ru-based photosensitizers.

Heterogeneous binding of dioxygen: EPR and DFT evidence for side-on nickel(II)-superoxo adduct with unprecedented magnetic structure hosted in MFI zeolite.

This article reports on the activation of dioxygen on nickel(I) dispersed inside the nanopores of the ZSM-5 zeolite, which can be regarded as a heterogeneous mimetic system (zeozyme) for Ni-bearing enzymes. The side-on η(2)-coordination of the resulting nickel-bound superoxo adduct was ascertained by detailed analysis of the EPR spectra of both (16)O(2) and (17)O(2) species supported by computer simulations of the spectra and relativistic DFT calculations of the EPR signatures. Molecular analysis of the g and A((17)O) tensors (g(xx) = 2.0635, g(yy) = 2.0884, g(zz) = 2.1675; |A(xx)| ≈ 1.0 mT, |A(yy)| = 5.67 mT, |A(zz)| ≈ 1.3 mT) and quantum chemical modeling revealed an unusual electronic and magnetic structure of the observed adduct (with g(zz)(g(max)) > g(yy)(g(mid)) > g(xx)(g(min)) and the largest O-17 hyperfine splitting along the g(mid) direction) in comparison to the known homogeneous and enzymatic nickel-superoxo systems. It is best described as a mixed metalloradical with two supporting oxygen donor ligands and even triangular spin-density redistribution within the η(2)-{NiO(2)}(11) magnetophore. The semioccupied molecular orbital (SOMO) is constituted by highly covalent δ overlap between the out-of-plane 2p(π(g)*) MO of dioxygen and the 3d(x(2)(-y(2))) MO of nickel. By means of the extended transition state-natural orbitals for the chemical valence approach (ETS-NOCV), three distinct orbital channels (associated with σ, π, and δ overlap) of congruent and incongruent charge and spin density flows within the η(2)-{NiO(2)}(11) unit, contributing jointly to activation of the attached dioxygen, were identified. Their individual energetic relevance was quantified, which allowed for explaining the oxygen binding mechanism with unprecedented accuracy. The nature and structure sensitivity of the g tensor was rationalized in terms of the contributions due to the magnetic field-induced couplings of the relevant molecular orbitals that control the g-tensor anisotropy. The calculated O-17 hyperfine coupling constants correspond well with the experimental parameters, supporting assignment of the adduct. To the best of our knowledge, the η(2)-{NiO(2)}(11) superoxo adducts have not been observed yet for digonal mononuclear nickel(I) centers supported by oxygen donor ligands.

Spin ground state and magnetic properties of cobalt(II): relativistic DFT calculations guided by EPR measurements of bis(2,4-acetylacetonate)cobalt(II)-based complexes.

The spin ground state of the core ion and structure of the bis(2,4-acetylacetonate)cobalt(II) model complex and its synthetic aqua and ethanol derivatives, Co(acac)(2)L(n), (L = EtOH, H(2)O), were examined by means of density functional theory (DFT) calculations supported by electron paramagnetic resonance (EPR) measurements. Geometry optimizations were carried out for low-spin (doublet) and high-spin (quartet) states. For the Co(acac)(2) complex two possible conformations, a square-planar and a tetrahedral one, were taken into account. For all structures relative energies were calculated with both "pure" and hybrid functionals. The calculated data were complemented with the results of the EPR investigations carried out at liquid helium temperature, allowing for definite assignment of the high-spin state for the Co(acac)(2)(EtOH)(2) complex. However, because of the unresolved spectral features, only effective g-values could be assessed, whereas the zero-field splitting parameters (ZFS) were calculated by means of the spin-orbit mean field (SOMF) relativistic DFT method for which direct spin-spin (SS) and spin-orbit coupling (SOC) contributions were quantified.

Magnetic Properties of Monomer and Dimer Tetrahedral VO(x) Entities Dispersed on Amorphous Silica-based Materials: Prediction of EPR Parameters from Relativistic DFT Calculations and Broken Symmetry Approach to Exchange Couplings.

Molecular structures of the isolated tetrahedral oxovanadium(IV) and bridged µ-oxo-divanadium(IV) complexes hosted by the clusters mimicking surfaces of amorphous silica-based materials were investigated using density functional theory (DFT) calculations. Principal values of the g and A tensors for the monomer vanadyl species were obtained using the coupled-perturbed DFT level of theory and the spin-orbit mean-field approximation (SOMF). Magnetic exchange interaction for the µ-oxo bridged vanadium(IV) dimer was investigated within the broken symmetry approach. An antiferromagnetic coupling of the individual magnetic moments of the vanadium(IV) centers in the [VO-O-VO](2+) bridges was revealed and discussed in detail. The coupling explains pronounced decrease of the electron paramagnetic resonance signal (EPR) intensity, observed for the reduced VO(x)/SiO(2) samples with the increasing coverage of vanadia, in terms of transformation of the paramagnetic monomer species into the dimers with S = 0 ground state.

Enhanced adsorption and degradation of methylene blue over mixed niobium-cerium oxide - Unraveling the synergy between Nb and Ce in advanced oxidation processes.

Formation of reactive oxygen species (ROS) via H2O2 activation is of vital importance in catalytic environmental chemistry, especially in degradation of organic pollutants. A new mixed niobium-cerium oxide (NbCeOx) was tailored for this purpose. A thorough structural and chemical characterization of NbCeOx along with CeO2 and Nb2O5 reference materials was carried out using TEM/STEM/EDS, SEM, XRD, XPS, EPR, UV-vis and N2 physisorption. The ability of the catalysts to activate H2O2 towards ROS formation was assessed on the basis of EPR and Raman measurements. Catalytic activity of the oxides was evaluated in degradation of methylene blue (MB) as a model pollutant. Very high activity of NbCeOx was attributed to the mixed redox-acidic nature of its surface, which originated from the synergy between Nb and Ce species. These two properties (redox activity and acidity) ensured convenient conditions for efficient activation of H2O2 and degradation of MB. The activity of NbCeOx in MB degradation was found 3 times higher than that of the commercial Nb2O5 CBMM catalyst and 240 times higher than that of CeO2. The mechanism of the degradation reaction was found to be an adsorption-triggered process initiated by hydroxyl radicals, generated on the surface via the transformation of O2-•/O22-.

Spectroscopic CW-EPR and HYSCORE investigations of Cu(2+) and O(2)(-) species in copper doped nanoporous calcium aluminate (12CaO.7Al(2)O(3)).

Continuous wave (CW) and pulse electron paramagnetic resonance in a variant of hyperfine sublevel correlation spectroscopy (HYSCORE) were used for obtaining structural information concerning speciation and local environment of alien Cu(2+) and native O(2)(-) ions encaged in copper doped nanoporous 12CaO.7Al(2)O(3) (mayenite). The samples were prepared by a solid-state reaction and characterized by means of XRD, SEM, and Raman techniques. X-Band CW-EPR spectra showed that three different Cu(2+) species together with paramagnetic extraframework O(2)(-) anions were present in the mayenite sample, whereas extraframework OH(-) anions were revealed by Raman spectroscopy. (27)Al HYSCORE provided evidence for the interaction of Cu(2+) ions with the mayenite framework. Superhyperfine interaction of the Cu(2+) ions with proximal (d(Cu-OH) = 2.4 A) and distal (d(Cu-OH) = 5.0 A) OH(-) anions, located in the same and the nearby cage, respectively, was resolved by means of (1)H HYSCORE spectra. A different situation held for the encaged O(2)(-) radicals found to be sitting on the Ca(2+) ions. They exhibited only a weak superhyperfine interaction of 1 MHz with the (27)Al(3+) framework ions.

Correction: Stability of Cu(II) complexes with FomA protein fragments containing two His residues in the peptide chain.

Correction for 'Stability of Cu(ii) complexes with FomA protein fragments containing two His residues in the peptide chain' by Monika Katarzyna Lesiów et al., Metallomics, 2019, 11, 1518-1531, DOI: 10.1039/C9MT00131J.

Stability of Cu(ii) complexes with FomA protein fragments containing two His residues in the peptide chain.

The coordination of Cu(ii) ions by the Ac-KGHGNGEEGTPTVHNE-NH2 (1L) peptide - a FomA protein fragment of Fusobacterium nucleatum- and its cyclic analogue: cyclo(KGHGNGEEGTPTVHNE) (2L) was studied by potentiometric titration, spectroscopic methods (UV-Vis, CD, EPR) and mass spectrometry (MS). Both the ligands contain two histydyl residues located in the third and fourteenth position of the peptide chain. For the 1L and 2L ligands mono- and dinuclear complexes were identified and studied in an aqueous solution. At the pH range characteristic of the intestinal environment (5.5-7.5), copper(ii) complexes were identified and their formation constants were determined. The same forms of the complexes with respectively the linear peptide and the cyclic peptide show similar stability, but greater than that reported in the literature for complexes with the same coordination mode. Moreover, the 1L peptide and its complex exhibit an α-helix structure, whereas the 2L peptide adopts this secondary structure only after coordination with the metal ion.

ROS-mediated lipid peroxidation as a result of Cu(ii) interaction with FomA protein fragments of F. nucleatum: relevance to colorectal carcinogenesis.

The ability of the studied FomA protein fragments of Fusobacterium nucleatum (Fn) with copper(ii) ions (Cu(ii)-Ac-KGHGNGEEGTPTVHNE-NH2 (1Cu) and its cyclic analogue Cu(ii)-cyclo(KGHGNGEEGTPTVHNE) (2Cu)) to induce reactive oxygen species (ROS) generation, as a result of red-ox processes, was determined by UV-Vis, luminescence methods, spin trapping and cyclic voltamperometry. The contribution of 1O2 and ˙OH to DNA degradation was proved using gel electrophoresis. Furthermore, the pronounced generation of ROS by mouse colon carcinoma cells (CT26) stimulated by both copper(ii) complexes was confirmed. A fluorescence method allowed the total amounts of ROS generated inside the CT26 cells to be detected, while the spin trapping technique proved that free radicals mainly attached to the membrane surface. These last results are in agreement with the data obtained from the ICP-MS method, which demonstrates that 1Cu and 2Cu complexes are not efficiently accumulated inside the cell. Furthermore, the role of ROS in lipid peroxidation was established. The above-mentioned factors may clearly indicate the contribution of ROS generated by the studied copper(ii) complexes to colonic cell damage, which can lead to a carcinogenesis process. This study may be an important step to recognize and understand the mechanism of colon cancer initiation.