Copper(II)-Assisted Degradation of Pheophytin a by Reactive Oxygen Species.

The central ion Mg2+ is responsible for the differences between chlorophyll a and its free base in their reactivity toward metal ions and thus their resistance to oxidation. We present here the results of spectroscopic (electronic absorption and emission, circular dichroism, and electron paramagnetic resonance), spectroelectrochemical, and computational (based on density functional theory) investigations into the mechanism of pheophytin, a degradation that occurs in the presence of Cu ions and O2. The processes leading to the formation of the linear form of tetrapyrrole are very complex and involve the weakening of the methine bridge due to an electron withdrawal by Cu(II) and the activation of O2, which provides protection to the free ends of the opening macrocycle. These mechanistic insights are related to the naturally occurring damage to the photosynthetic apparatus of plants growing on metal-contaminated soils.

Bispalladium(II) Complexes of di-p-Pyrirubyrin Derivatives as Promising Near-Infrared Photoacoustic Dyes.

The insertion of palladium(II) into di-p-pyrirubyrin results in mutually convertible bimetallic complexes. Post-synthetic functionalization of one of them yielded bispalladium(II) dioxo-di-p-pyrirubyrin and, after demetallation, dioxo-di-p-pyrirubyrin, introducing for the first time the α,ß'-pyridin-2-one unit into the macrocyclic frame. Bispalladium(II) di-p-pyrirubyrin 6, bispalladium(II) dioxo-di-p-pyrirubyrin 9, and dioxo-di-p-pyrirubyrin 10 absorb and emit light around 1000 nm and are characterized by high photostability. Thus, they are promising candidates for near-infrared photoacoustic dyes, ideally targeting (9) the wavelength of Yb-based fiber lasers. The incorporation of an α,ß'-pyridine moiety into expanded porphyrins opens a highly interesting area of research due to the attractive optical and coordination properties of the resulting molecules.

Chlorophyll a π-Cation Radical as Redox Mediator in Superoxide Dismutase (SOD) Mimetics.

The extensive speciation of copper(II) chloride in organic solvents varies with concentration, temperature, pressure and oxygen content, providing the ability to switch between different chlorophyll transmetalation pathways. We found that one of them is exceptionally suitable for the formation and stabilisation of the chlorophyll π-cation radical. This is due to unique redox cycling, which is coupled to the generation and transformation of various reactive oxygen species. In the presence of a proton donor, our system shows behavior which resembles that of superoxide dismutase (SOD). Regardless of light, chlorophyll acts as an electron transfer mediator.

Mechanistic Studies on the Reaction between Aquacobalamin and the HNO Donor Piloty's Acid over a Wide pH Range in Aqueous Solution.

Detailed kinetic and mechanistic studies have been carried out on the reaction between aquacobalamin/hydroxocobalamin (CblOH2+/CblOH) and nitroxyl (HNO) generated by Piloty's acid (PA, N-hydroxybenzenesulfonamide) over a wide pH range (3.5-13). The resulting data showed that in a basic solution HNO can react with hydroxocobalamin to form nitrosylcobalamin despite the inert nature of CblOH. It was shown that at low PA concentrations the rate-determining step is the decomposition of PhSO2NHO- to release HNO, whereas the reaction between CblOH and HNO becomes the rate-determining step at high PA concentrations. Data from kinetic studies on the reaction of CblOH with an excess of HNO enabled us to experimentally determine the pKa(HNO) value from initial rate data as a function of pH, giving pKa(HNO) = 11.47 ± 0.04. An especially interesting observation was made in the neutral pH range, where PA is stable and does not produce HNO. Under such conditions, rapid formation of CblNO was observed in the studied system. The obtained data suggest that CblOH2+ reacts directly with PA to form a Piloty's acid-bound cobalamin intermediate, which deprotonates rapidly at neutral pH followed by rate-determining S-N bond cleavage to give CblNO and release PhSO2-.

Ligand-Tuning of the Stability of Pd(II) Conjugates with Cyanocobalamin.

Besides the well-known functions performed by vitamin B12 (CblCN) in biochemical processes of the human body, an increasing interest has been raised by the possibility of its use as a transmembrane drug carrier, capable, among others, of enhancing the accumulation of inorganic cytostatics in cancer cells. The present study was aimed at determining the possibility of the formation of CblCN conjugates with Pd(II) complexes. A key aspect was their stability, which we attempted to tune by appropriate choice of ligands. Syntheses, spectroscopic analysis of postreaction systems and kinetic investigations of conjugate formation reactions, have been complemented by DFT modelling. The obtained results showed that ligand charge, geometry and electron affinity may have a significant impact on carrier binding and release leading to the activation of the Pd(II) complex. This provides a rationale to expect that with appropriate composition of the coordination sphere, it will be possible to extend the spectrum of less toxic inorganic chemotherapeutics.

High-Pressure Mechanistic Insight into Bioinorganic NO Chemistry.

Pressure is one of the most important parameters controlling the kinetics of chemical reactions. The ability to combine high-pressure techniques with time-resolved spectroscopy has provided a powerful tool in the study of reaction mechanisms. This review is focused on the supporting role of high-pressure kinetic and spectroscopic methods in the exploration of nitric oxide bioinorganic chemistry. Nitric oxide and other reactive nitrogen species (RNS) are important biological mediators involved in both physiological and pathological processes. Understanding molecular mechanisms of their interactions with redox-active metal/non-metal centers in biological targets, such as cofactors, prosthetic groups, and proteins, is crucial for the improved therapy of various diseases. The present review is an attempt to demonstrate how the application of high-pressure kinetic and spectroscopic methods can add additional information, thus enabling the mechanistic interpretation of various NO bioinorganic reactions.

Nitrosyl- versus nitroxyl-cobalamin?

The Commentary is in answer to the comment of a reader that objected against the use of the term 'nitroxylcobalamin' in two recent reports in JBC from our group. We use this opportunity to explain to the reader where this terminology originated from.

Can nitrocobalamin be reduced by ascorbic acid to nitroxylcobalamin? Some surprising mechanistic findings.

Despite detailed studies on nitroxylcobalamin (CblNO) formation, the possible intracellular generation of CblNO via reduction of nitrocobalamin (CblNO2) remains questionable. To study this further, spectroscopic studies on the reaction of CblNO2 with the intracellular antioxidant ascorbic acid (HAsc-) were performed in aqueous solution at pH < 5.0. It was found that nitroxylcobalamin is the final product of this interaction, which is not just a simple reaction but a rather complex chemical process. We clearly show that an excess of nitrite suppresses the formation of CblNO, from which it follows that ascorbic acid cannot reduce coordinated nitrite. We propose that under the influence of ascorbic acid, nitrocobalamin is reduced to Cbl(II) and nitric oxide (·NO), which can subsequently react rapidly to form CblNO. It was further shown that this system requires anaerobic conditions as a result of the rapid oxidation of both Cbl(II) and CblNO.

Mechanistic information on the nitrite-controlled reduction of aquacob(III)alamin by ascorbate at physiological pH.

The interaction with nitric oxide (NO) is an important aspect of the biological activity of vitamin B12 (Cbl). Whereas the formation of nitroxylcobalamin (CblNO) via the binding of NO to reduced CblCo(II) has been studied in detail before, the possible intracellular formation of CblNO via reduction of nitrocobalamin (CblNO2) is still questionable. To study this further, spectroscopic and kinetic studies on the reaction of CblNO2 with the intracellular antioxidant ascorbic acid (Asc) were performed in aqueous solution at the physiological pH of 7.2. It was found that the redox pathway of this reaction requires anaerobic conditions as a result of the rapid re-oxidation of reduced CblCo(II). In the studied system, both CblOH2 and CblNO2 are reduced to CblCo(II) by ascorbate at pH 7.2, the CblOH2 complex being two orders of magnitude more reactive than CblNO2. Clear evidence for redox cycling between CblOH2/CblNO2 and CblCo(II) under aerobic conditions was observed as an induction period during which all oxygen was used prior to the formation of CblCo(II) in the presence of an excess of ascorbate. No evidence for the intermediate formation of CblNO or NO radicals during the reduction of CblNO2 could be found. Nitrocob(III)alamin can be reduced by ascorbic acid under physiological conditions. The products of the reaction are cob(II)alamin and nitrite ion. This reaction is ca. 200 times slower than the one involving aquacob(III)alamin.

The lowest triplet states of bridged cis-2,2'-bithiophenes - theory vs. experiment.

Theoretical methods that were previously used to give a good quantitative description of the 3(1)Bu state of trans-2,2'-bithiophene are applied to characterize the lowest triplet states of three bridged cis-2,2'-bithiophenes: 3,3'-cyclopentadithiophene (CPDT), 3,3'-dithienylpyrrole (DTP), and 3,3'-dithienylthiophene (DTT). By obtaining highly accurate reproductions of the phosphorescence spectra of all three compounds, we rationalize the observed vibronic activity, further explore the performance of the applied theoretical methods, and address the quality of the reported experimental spectra. Over the course of this study we have, first, characterized the changes in the electronic structures between the ground state and the lowest triplet state and, second, expressed the related geometrical differences in terms of the Huang-Rhys factors. The Huang-Rhys factors have then been used to generate theoretical emission spectra with vibronic resolution. The applied procedure has yielded quantitative reproductions of the previously reported experimental phosphorescence spectra of DTT and DTP. The experimental spectrum of CPDT, on the other hand, turned out to be considerably narrower and intensity-deficient in its low energy region when compared with the theoretical results. Our experimental reinvestigation of the CPDT phosphorescence has given a refined spectrum that is significantly wider than the previously reported one, and is in nearly quantitative agreement with the theoretical prediction. This enabled us to attribute the observed discrepancy to an experimental artifact associated with the sensitivity characteristics of the commonly used photomultipliers.

Joint theoretical and experimental study on the phosphorescence of 2,2'-bithiophene.

Theoretical investigation into vibronic activity observed in the phosphorescence spectrum of 2,2'-bithiophene (2T) has revealed unexpectedly large discrepancies between the theoretical predictions and the available experimental spectrum. Theoretical spectra, obtained using the results of the high-level quantum chemistry calculations and the well founded model for vibronic activity, are considerably wider than the experimental one. The energy of the onset (corresponding to the adiabatic transition energy) is quantitatively reproduced, but the centers of gravity of the simulated spectral bands (vertical emission energies) are considerably red-shifted with respect to experiment. Critical reconsideration and tests of the approximations underlying the theoretical approach have failed to pinpoint the source of the observed discrepancies. In the following experimental reinvestigation of the phosphorescence of 2T we have observed that unusually large concentration of the chromophore and careful recalibration of the detector prior to measurements have been essential to collect a good quality spectrum. Incidentally, being considerably different from the previously reported one, the newly reported phosphorescence spectrum of 2T is in nearly quantitative agreement with the theoretical predictions.

Conjugation Paths in Oxatriphyrin(2.1.1) - C2 Bridge Modifications.

An efficiency of delocalization in strongly conjugated systems remains an important factor crucial for modulation of the optical properties directly correlated with its range. An ortho-substituted phenylene derivative bearing electron donating/accepting functionality was built-in a fully unsaturated macrocyclic system with a global delocalization of a diatropic and/or a paratropic current. A precisely located structural modification influence observed behaviour in spectroscopic parameters that are only slightly recognizable in 4n+2 systems but showing a significant influence on the reduced derivatives with a contribution of 4n π-electrons delocalization path.

Structural and electronic effects in the metalation of porphyrinoids. Theory and experiment.

The structure-reactivity relationships in metalation reactions of porphyrinoids have been studied using experimental and theoretical methods. A series of eight porphyrinoic ligands, derivatives of chlorophylls, was prepared in which both the peripheral groups and the degrees of saturation of the macrocycle were systematically varied. To reveal the solvent and structural factors which control the interactions of these macroligands with metal centers, their interactions with reactive Zn(2+) and inert Pt(2+) ions were investigated using absorption spectroscopy. In parallel, quantum chemical calculations (density functional theory, DFT) were performed for the same set of molecules to examine the influence of structural and electronic factors on the energy of the frontier orbitals, the nucleophilicity/electronegativity of the macrocycle, its hardness, and conformation. These static descriptors of chemical reactivity, relevant to metalation reactions, were verified against the results obtained in the experimental model. The experimentally obtained kinetic data clearly show that the solvent has a crucial role in the activation of the incoming metal center. In terms of chelator structure, the largest effects concern the size of the delocalized pi-electron system and the presence of side groups. Both the DFT calculations and experimental results show the strong influence of the macrocycle rigidity and of the peripheral groups on the chelating ability of porphyrinoids. In particular, the peripheral functionalization of the macrocyclic system seems to drastically reduce its reactivity toward metal ions. The effect of peripheral groups is two-fold: (i) a lower electron density on the core nitrogens, and (ii) increased rigidity of the macrocycle. The outcomes of the theoretical and experimental analyses are discussed also in terms of their relevance to the mechanism of biological metal insertion in the biosynthesis of heme and chlorophyll.

Application of high pressure laser flash photolysis in studies on selected hemoprotein reactions.

This article focuses on the application of high pressure laser flash photolysis for studies on selected hemoprotein reactions with the objective to establish details of the underlying reaction mechanisms. In this context, particular attention is given to the reactions of small molecules such as dioxygen, carbon monoxide, and nitric oxide with selected hemoproteins (hemoglobin, myoglobin, neuroglobin and cytochrome P450(cam)), as well as to photo-induced electron transfer reactions occurring in hemoproteins (particularly in various types of cytochromes). Mechanistic conclusions based on the interpretation of the obtained activation volumes are discussed in this account.

Understanding chlorophylls: central magnesium ion and phytyl as structural determinants.

Phytol, a C20 alcohol esterifying the C-17(3) propionate, and Mg2+ ion chelated in the central cavity, are conservative structural constituents of chlorophylls. To evaluate their intramolecular structural effects we prepared a series of metal- and phytyl-free derivatives of bacteriochlorophyll a and applied them as model chlorophylls. A detailed spectroscopic study on the model pigments reveals meaningful differences in the spectral characteristics of the phytylated and non-phytylated pigments. Their analysis in terms of solvatochromism and axial coordination shows how the central Mg and phytyl residue shape the properties of the pigment. Surprisingly, the presence/absence of the central Mg has no effect on the solvatochromism of (bacterio)chlorophyll pi-electron system and the hydrophobicity of phytyl does not interfere with the first solvation shell of the chromophore. However, both residues significantly influence the conformation of the pigment macrocycle and the removal of either residue increases the macrocycle flexibility. The chelation of Mg has a flattening effect on the macrocycle whereas bulky phytyl residue seems to control the conformation of the chromophore via steric interactions with ring V and its substituents. The analysis of spectroscopic properties of bacteriochlorophyllide (free acid) shows that esterification of the C-17(3) propionate is necessary in chlorophylls because the carboxyl group may act as a strong chelator of the central Mg. These observations imply that the truncated chlorophylls used in theoretical studies are not adequate as models of native chromophores, especially when fine effects are to be modeled.

Hemicyanine dyes derived from 2,3,3-trimethyl-3H-indolium as candidates for non-covalent protein probes.

Non-covalent interaction of hemicyanine dyes, derivatives of 2,3,3-trimethyl-3H-indolium with bovine serum albumin has been studied by spectral method. For this purpose, three hemicyanine dyes containing N-(2-carboxyethyl)-2,3,3-trimethyl-3H-indolium moiety were synthesized and their UV/Vis and fluorescence spectra, aggregation, photostability and association with bovine serum albumin were studied. The hemicyanine dyes with 2-ethylcarboxylic group was found to interact with bovine serum albumin, which is probably due to negative charge on the dye molecule at the expense of the carboxylic group and the ability to form hydrogen bonds with albumin.

Fine tuning of copper(II)-chlorophyll interactions in organic media. Metalation versus oxidation of the macrocycle.

The nature of chlorophyll interactions with copper(II) ions varies considerably in organic solvents, depending on the dominant coordinative form. Besides formation of the metallo tetrapyrrolic complex, Cu(II) ions can cause oxidation of the pigment, reversible or irreversible, which can lead to the destruction of the macrocyclic structure. All these reaction types can be distinguished within a quite narrow range of reaction conditions. The ability to form new metallo derivatives in either metalation or transmetalation reactions is obviously limited by the concentration of the potential oxidant, but can be secured below this level via suitable composition of the reaction system. The decisive factor in the selection of a specific reaction pathway is the presence of a potential ligand that can affect the reactivity of Cu(II) for example by shifting its redox potential. Spectroscopic and electrochemical studies were performed in order to determine the predominant species of Cu(II) in methanol, nitromethane and acetonitrile in the presence of chloride and acetate ions, as well as to assign their appropriate oxidizing ability. This allowed us to estimate the boundary conditions for the electron transfer processes in chlorophyll-Cu(II) systems. Chlorophyll and its free base can undergo both types of electron transfer processes, however, they reveal different susceptibilities that make this class of ligands quite versatile markers in tuning the reactivity of metal ions in solutions.

Exploring novel modified vitamin B12 as a drug carrier: forecast from density functional theory modeling.

Three non-native derivatives of vitamin B12 with imidazole, ethylenediamine, and pyrazine as cobalt(III) ß-ligands were characterized by applying the BP/def2-TZVP density functional method. The binding of all three ligands is thermodynamically favorable. It is proposed that their synthesis might be possible from aquacobalamin as a starting form of vitamin B12, as has been done in the case of an imidazole derivative of B12 (Hannibal et al. Inorg. Chem. 2007, 46, 3613-3618). Furthermore, the possibility of the formation of their conjugates with cisplatin is investigated. The proposed ß-ligands may serve as bridging ligands, binding to the platin ion as N-donors. In parallel, the calculations are done for the previously synthetized B12-cisplatin adduct with CN(-) as a bridging ligand and are compared with available experimental data, allowing assessment of the applied computational protocol. A good agreement between the computed and experimental structural parameters is obtained. In each of the studied structures, the Co-ß-ligand bond is weaker than the Pt-ß-ligand bond.

Interplay between acetate ions, peripheral groups, and reactivity of the core nitrogens in transmetalation of tetrapyrroles.

The mechanism of acetate-assisted transmetalation of tetrapyrroles was investigated in a model system consisting of chlorophyll a and copper(II) acetate in organic solvents by using a spectroscopic and kinetic approach. Surprisingly, acetate ions bind to the central Mg in chlorophyll much more strongly than do acetonitrile, methanol and even pyridine, one of the best ligands in chlorophyllic systems. This exceptionally strong non-symmetrical axial ligation of the central Mg by acetate causes its out-of-plane displacement and deformation of the tetrapyrrole ring, thus facilitating the interaction with an incoming CuII complex. This mechanism is controlled by a keto-enol tautomerism of the chlorophyll isocyclic ring. Additionally, depending on solvent, acetate activates the incoming metal ions. These new insights allow to suggest a mechanism for the acetate method of metal exchange in tetrapyrrolic macrocycles, which resembles biological insertion of metal ions into porphyrins. It also provides a guideline for the design of more efficient methods for the metalation of porphyrins and related macrocycles.