Incorporation Of Copper Ions Research Articles

Exploiting Distinct Molecular Architectures of Ultrathin Films Made with Iron Phthalocyanine for Sensing

The possibility of generating distinct film properties from the same material is crucial for a number of applications, which can only be achieved by controlling the molecular architecture. In this paper we demonstrate as a proof-of-principle that ultrathin films produced from iron phthalocyanine (FePc) may be used to detect trace amounts of copper ions in water, where advantage was taken of the cross sensitivity of the sensing units that displayed distinct electrical properties. The ultrathin films were fabricated with three methods, namely physical vapor deposition (PVD), Langmuir-Blodgett (LB), and electrostatic layer-by-layer (LbL) techniques, where for the latter tetrasulfonated phthalocyanine was used (FeTsPc). PVD and LB films were more homogeneous than the LbL films at both microscopic and nanoscopic scales, according to results from micro-Raman spectroscopy and atomic force microscopy (AFM), respectively. From FTIR spectroscopy data, these more homogeneous films were found to have FePc molecules oriented preferentially, tilted in relation to the substrate surface, while FeTsPc molecules were isotropically distributed in the LbL films. Impedance spectroscopy measurements with films adsorbed onto interdigitated gold electrodes indicated that the electrical response depends on the type of film-forming method and varies with incorporation of copper ions in aqueous solutions. Using principal component analysis (PCA), we were able to exploit the cross sensitivity of the sensing units and detect copper ions (Cu(2+)) down to 0.2 mg/L, not only in ultrapure water but also in distilled and tap water. This level of sensitivity is sufficient for quality control of water for human consumption, with a fast, low-cost method.

Effect of Copper Doping on the Crystal Structure and Morphology of 1D Nanostructured Manganese Oxides

We have tried to control the aspect ratio and physicochemical properties of 1D nanostructured manganese oxides through copper doping. Copper-doped manganese oxide nanostructures have been synthesized by one-pot hydrothermal treatment for the mixed solution of permanganate anions and copper cations. According to powder X-ray diffraction and electron microscopic analyses, all the present materials commonly crystallize with alpha-MnO2-type structure but their aspect ratio decreases significantly with increasing the content of copper. Such a variation of crystallite dimension is attributable to the limitation of crystal growth by the incorporation of copper ions. X-ray absorption spectroscopic studies at Mn K- and Cu K-edges clearly demonstrate that the average oxidation state of manganese ions is increased by the substitution of divalent copper ions. Electrochemical measurements reveal the improvement of the electrode performance of nanostructured manganate upon copper doping, which can be interpreted as a result of the decrease of aspect ratio and the increase of Mn valence state. From the present experimental findings, it becomes certain that the present Cu doping method can provide an effective way of controlling the crystal dimension and electrochemical property of 1D nanostructured manganese oxide.

Cox17, a copper chaperone for cytochrome c oxidase: Expression, purification, and formation of mixed disulphide adducts with thiol reagents

Copper chaperone for cytochrome c oxidase (Cox17) is a 7 kDa copper-binding protein, which facilitates incorporation of copper ions into Cu A site of cytochrome c oxidase. Cox17 contains six conserved Cys residues and occurs in three different oxidative states, which display different metal-binding properties and stability. In the present study, we have elaborated technologies for production of partially oxidized human recombinant Cox17 in a bacterial expression system and purification of fully oxidized Cox17. For this purpose we used Escherichia coli Origami™ strain, which is deficient in thioredoxin and thioredoxin reductase systems and allows formation of disulfide bonds in cytoplasmic proteins. Fully oxidized Cox17 was purified by a simplified two-step procedure including gel filtration and cation exchange chromatography. By using mass spectrometry we demonstrated that application of 2-mercaptoethanol (2-ME) during purification leads to formation of its mixed disulfide adducts with Cox17. Moreover, partially reduced Cox17 can form mixed disulfide adducts also with the cellular reducing agent glutathione, which abolishes copper-binding ability of partially reduced Cox17.

Distributions of copper and CuBr in CuBr nanocrystallite-dispersed glasses prepared by copper staining

CuBr nanocrystallite-dispersed glasses were prepared by incorporation of copper into bromide ion-containing borosilicate glass using the technique of copper staining. The copper ion incorporation process was mainly controlled by ionic diffusion from the surface to the interior of the glass. The depth profiles of Cu and CuBr concentration were examined by energy dispersive X-ray analysis and by reference to the change in absorption intensity assigned to the CuBr exciton band along the depth. While the Cu concentration was found to decrease monotonically, the CuBr concentration profile showed a maximum at a distance of 10–50μm from the glass surface. Although the depth reached by the copper ions became greater with increasing heat-treatment time, the depth at which CuBr was precipitated was found to be saturated. This means that regions were found in the glasses in which no CuBr crystallites precipitated, although migration of Cu ions to these regions had taken place.

Cu-doped TiO2 systems with improved photocatalytic activity

Photocatalytic oxidation of phenol was performed over copper doped TiO2 prepared by a sol–gel method. Different preparation methods were followed and a comparison with undoped system was also made. Wide structural and surface characterization of catalysts was carried out in order to establish a correlation between the effect of sulphuric and nitric acids present in the initial solution and the Cu–TiO2 photocatalytic properties. The presence of sulphuric acid clearly stabilizes Cu–TiO2 and TiO2 structure and surface against sintering, maintaining anatase phase and relatively high surface area values with respect non sulphated Cu–TiO2 or TiO2. Best photocatalytic behaviour is found for sulphated TiO2, as previously reported. In addition, incorporation of copper ions into the structure seems to enhance the photoactivity of the system for acidified systems, being the sulphated one the most favourable photocatalyst. The optimum metal loading is found for 0.5M% of copper ion. A possible explanation of this photocatalytic improvement might be related to the stabilization of Cu2O species in doped TiO2 prepared in the presence of sulphuric acid. This stabilization could be related to the presence of oxygen vacancies generated in the preparation procedure using sulphuric acid.

Synthesis, characterization, and properties of poly(m‐aminophenyl propionic acid) and its copolymers with aniline

AbstractThe synthesis of poly(m‐aminophenyl propionic acid) (PAPPA) was carried out by polymerization in different aqueous media, from the respective monomer, using ammonium persulfate as oxidizing agent. The copolymer of aniline and m‐aminophenyl propionic acid was also synthesized. The polymers were characterized by FTIR, UV‐vis, X‐ray electron spectroscopy, nuclear magnetic resonance spectroscopy, elemental analysis, thermal stability analysis (TGA), and electrical conductivity. PAPPA showed a high doping by HCl and H2SO4. If the composition of the comonomer acid decreases, the incorporation of copper ion and the electrical conductivity are increased. Thermal stability is favored by an increase of the content of aniline and copper ions. The polymeric structures have amine and imine groups. The last units are preferentially doped. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 90: 706–715, 2003

A Pivotal Role of Zn-Binding Residues in the Function of the Copper Chaperone for SOD1

A Cu chaperone for SOD1 (CCS) is required for the incorporation of copper ion into the protein. To investigate the roles of the conserved metal-binding residues in CCS, we introduced amino acid substitutions into human CCS and examined the function of the mutant CCS by transforming a mutant yeast strain, SY2950, which lacks the lys7 gene, a yeast orthologue of the mammalian CCS. Mutant CCS in which amino acid residues His147 and Asp167 were substituted by Ala exhibited a decreased ability to complement the growth of SY2950 under Lys-deficient conditions. This is because the mutations made the human CCS function in a less efficient manner, especially under metal-restricted conditions, leaving Cu,Zn-SOD in an apo-form. Since the His and Asp residues are both responsible for binding Zn which would serve to maintain the folded structure, the structural integrity supported by the coordinated Zn ion would be essential for CCS function.

A Gain of Superoxide Dismutase (SOD) Activity Obtained with CCS, the Copper Metallochaperone for SOD1

The incorporation of copper ions into the cytosolic superoxide dismutase (SOD1) is accomplished in vivo by the action of the copper metallochaperone CCS (copper chaperone for SOD1). Mammalian CCS is comprised of three distinct protein domains, with a central region exhibiting remarkable homology (approximately 50% identity) to SOD1 itself. Conserved in CCS are all the SOD1 zinc binding ligands and three of four histidine copper binding ligands. In CCS the fourth histidine is replaced by an aspartate (Asp(200)). Despite this conservation of sequence between SOD1 and CCS, CCS exhibited no detectable SOD activity. Surprisingly, however, a single D200H mutation, targeting the fourth potential copper ligand in CCS, granted significant superoxide scavenging activity to this metallochaperone that was readily detected with CCS expressed in yeast. This mutation did not inhibit the metallochaperone capacity of CCS, and in fact, D200H CCS appears to represent a bifunctional SOD that can self-activate itself with copper. The aspartate at CCS position 200 is well conserved among mammalian CCS molecules, and we propose that this residue has evolved to preclude deleterious reactions involving copper bound to CCS.

Oxidation Of AI-Cu-Fe Quasicrystals

AbstractThe oxidation behavior of i-A163Cu25Fe12 at 800°C in air was investigated by means of TGA, XRD, SEM and TEM. In the beginning a homogeneous oxide layer is formed by the subsequent growth of metastable γ-Al2O3 and Θ-Al2O3. Nucleation of the thermodynamical stable α-Al2O3 occurs at the interface oxide/quasicrystal. The following growth of α-Al2O3 through the oxide layer leads to the formation of oxide nodules. The high growth rate of the α-Al2O3 can be explained by the incorporation of copper ions. The oxidation resistance of the quasicrystal is insufficient at high temperatures, because no protective oxide layer is formed. The high temperature oxidation behavior of Al-Cu-Fe quasicrystal and the aluminides β-FeAl and β-NiAl is compared regarding the oxidation rate, the oxide phases and the concentration changes in the material due to selective oxidation of aluminum.

On the incorporation of copper ions in heavy metal fluoride glasses

AbstractCu2+ ions incorporated into (ZBLAN) fluoride glasses reflect via distributions of spin coupling parameters local randomness of the glass matrix. Cw‐X‐band ESR measurements at 4.2 K applying different values of the microwave power as well as computer simulations have shown that even at very low copper concentrations (<0.6 mol%) the Cu2+ ions form point defects coexisting and weakly interacting with magnetically coupled species. The tendency of Cu2+ ions to form aggregates is remarkedly higher than in oxide glasses which corresponds to a nonuniform distribution of the paramagnetic centres in the glass matrix. After cyclic heating and cooling processes, recrystallization of the matrix represented by lowering the line widths, and resolving hyperfine splittings in the g∥ region could be observed.

Enrichment-dependent anodic oxidation of Zinc in Al-Zn Alloys

As part of a systematic study of anodic oxidation of aluminium alloys, focused on the importance of initial enrichment of thin layers of alloy to the oxidation of alloying elements at the alloy/film interface, the behaviour of Al/Zn alloys is considered, which is also of relevance to surface treatment of commercial aluminium alloys. The alloys investigated, by a combination of transmission electron microscopy and Rutherford backscattering spectroscopy, are bulk Al-0.90 at% Zn alloy, in the electropolished condition, and Al-0.23 at% Zn alloy, produced by magnetron sputtering. Following anodic oxidation at 50 Am −2, both alloys reveal zinc enrichments immediately beneath the amorphous anodic films, in thin layers of alloy which contain average concentrations of about 1.8 × 10 19 and 4.6 × 10 19 Zn atoms m −2, respectively. For the Al-0.9 at% Zn alloy, the enrichment is sufficient for zinc atoms to be oxidized at the alloy/film interface and, hence, for zinc ions to be incorporated into the mainly alumina film. The incorporated zinc ions migrate outwards in the anodic film about 2.3 times faster than Al 3+ ions. For the Al-0.23 at% Zn alloy, the enrichment is insufficient for oxidation of zinc atoms to proceed and, hence, no significant amount of zinc ions was detected in the film. The oxidation behaviour of zinc in Al-Zn alloys is generally similar to that of copper in dilute Al-Cu alloys, although the incorporation of zinc ions into the anodic films does not create the high population density of flaws found in films on Al-Cu alloys following incorporation of copper ions. Further, anodic film material on Al-Zn alloys becomes detached from the substrate at a particular condition of the alloy/film interface which, in the present work, occurs at an anodizing voltage of about 100 V for the Al-0.90 at% Zn alloy. Following detachment of the film, most of the zinc in the zincenriched alloy layer is lost from the exposed alloy surface and the enrichment reforms during subsequent regrowth of the anodic film.

The importance of surface treatment to the anodic oxidation behaviour of AlCu alloys

The anodic oxidation of Al-0.9 at%Cu alloy has been investigated, focusing on the effect of a mechanical polishing pre-treatment on incorporation of copper ions into the anodic film and the accumulation of copper atoms in the alloy just beneath the alloy/film interface. For the particular alloy and anodizing conditions, a relatively pure alumina film is formed to thickness of about 170 nm with associated progressive accumulation of copper atoms in the alloy at the alloy/film interface. The thickness of the copper-enriched layer is about 2 nm and the average composition approaches about 42 at%Cu. Only on achieving this level of enrichment are copper ions incorporated into the anodic film. The copper ions are more mobile than aluminium ions and hence, are able to reach the film/electrolyte interface during subsequent film growth.

Role of transit peptide sequence of plastocyanin for its expression, processing, and copper-binding activity in Escherichia coli.

Plastocyanin is a copper protein that functions as an electron carrier in the thylakoid lumen of the chloroplast. To characterize the transit peptide of plastocyanin and develop expression systems for it in Escherichia coli, three kinds of expression vectors which encode different size precursor plastocyanin molecules were constructed. Their expression, processing, and copper-binding activity have been examined. When the full-length cDNA encoding the precursor plastocyanin from Silene pratensis was expressed in E. coli, a large amount of precursor plastocyanin accumulated in insoluble aggregates. Its accumulation level was increased by the addition of copper ions. About six percent of precursor plastocyanin molecules were transported into the periplasmic space and processed to the mature protein. On the other hand, expression of the intermediate size cDNA, which contains the hydrophobic domain and basic amino acid of C-terminal transit peptide, caused exclusive translocation to the periplasmic space and correct processing to the mature size. The addition of copper ions increased the holo-protein content, but did not change the polypeptide content of mature plastocyanin, indicating that translocation and processing are independent of the incorporation of copper ions. The mature plastocyanin content corresponds to 8% (w/w) of the total E. coli protein content (123 mg per liter of culture). The purified mature holo-protein showed almost the same spectroscopic and kinetic properties as those of purified spinach plastocyanin. Expression of the cDNA encoding the mature polypeptide and two preceeding amino acid residues caused the accumulation of only a small amount of plastocyanin.(ABSTRACT TRUNCATED AT 250 WORDS)

Mutational study of Streptomyces tyrosinase trans-activator MelC1. MelC1 is likely a chaperone for apotyrosinase.

The melanin operon (melC) of Streptomyces antibioticus contains two genes, melC1 and melC2 (apotyrosinase). Our previous studies indicated that MelC1 forms a transient binary complex with the downstream apotyrosinase MelC2 to facilitate the incorporation of copper ion and the secretion of tyrosinase. In this study, we investigated the role of histidine residues in the function of MelC1 by examining a series of substitution or deletion mutants. Of eight mutants only the substitution of His-117 with Asp in the mutant M-117D rendered the complete abolishment of the intracellular tyrosinase activity in both Streptomyces and Escherichia coli. Replacement of His-102 by Leu in the mutant M-102L also caused a 64-70% reduction of tyrosinase activity in Streptomyces and E. coli. These two mutations also affected the secretion of both MelC1 and MelC2 proteins. In vitro copper activation of the purified MelC1.MelC2 binary complex from these two mutants regained only 20-30% tyrosinase activity of the wild type. Biochemical characterization of the tyrosinases from these two mutants revealed that they were different in several aspects. The intracellular tyrosinase activity in M-117D, but not in M-102L, could be partially reactivated by copper ion or by the cell extract containing MelC1. The copper content and the specific activity of the tyrosinase purified from the culture supernatant from M-117D were only 40% of those in wild type and M-102L. Additionally, fast protein liquid chromatography analysis indicated that in these two mutants the copper activation process was defective, very likely due to the incompetent MelC1.MelC2 binary complex formed: reduced association in M-117D and elevated association in M-102L. Furthermore, the conformation of MelC2 in the binary complex or in the mature enzyme form in wild type could be differentiated by the proteinase K digestion pattern, and so did the conformation of MelC2 found in those of M-102L, but not in M-117D mutant. Taken together, our results demonstrate that MelC1 is indispensable in the incorporation of copper ion into MelC2 apotyrosinase via a transient, competent binary complex formation, during which a conformational transition of MelC2 has occurred. This strongly suggests that MelC1 is a chaperone for the apotyrosinase MelC2.

Copper transfer and activation of the Streptomyces apotyrosinase are mediated through a complex formation between apotyrosinase and its trans-activator MelC1.

The melanin operon (melC) of Streptomyces antibioticus is composed of two genes that encode MelC1 and MelC2 proteins. MelC1 has been suggested as a trans-activator which can facilitate the incorporation of copper into the apotyrosinase (MelC2) (Lee, Y.-H. W., Chen, B.-F., Wu, S.-Y., Leu, W.-M., Lin, J.-J., Chen, C. W., and Lo, S. J. (1988) Gene (Amst.) 65, 71-81). However, the molecular mechanism of the trans-activation or copper-transfer process mediated through MelC1 to MelC2 is not clear yet. In this study, we found apotyrosinase in both the extracellular fraction and cell extract from cells grown in copper-deficient medium. Using gel-filtration and immunoaffinity chromatographies, we demonstrated that apotyrosinase (MelC2) formed a stable complex with MelC1 in the intra- and extracellular fractions. Furthermore, addition of copper ion to the complex generated tyrosinase activity. The MelC1-MelC2 complex was purified to near homogeneity by DE52 and phenyl-agarose chromatographies. In conjunction with fast protein liquid gel filtration chromatography and NH2-terminal sequencing analysis, the results indicated that the stoichiometric ratio of MelC1 and MelC2 in the purified complex was 1:1. Essentially no copper was found in the complex. Addition of copper ion to the purified complex resulted in incorporation of approximately 2 molecules of copper ion and the mature active tyrosinase was gradually released from the complex. Taken together, these results demonstrate that the molecular mechanism of activation of Streptomyces apotyrosinase by its trans-activator MelC1 is initially mediated via a binary complex formation between these two proteins, followed by incorporation of copper ion. This activation mechanism accounts for the essential role of MelC1 in the expression of melanin operon.

Synthetic porphyrins at interfaces; photosensitization and related reactions of atropisomers of o-substituted tetraphenylporphine derivatives in films, reversed micelles and membranes.

Abstract— Previous studies from our laboratories and elsewhere have shown that amides between fatty acids and the synthetic α, α, α, α ‐tetra (o‐aminophenyl)‐porphyrins exhibit good surfactant properties which facilitate their incorporation into structured assemblies characterized by hydrophobic‐hydrophilic compartmentalization. This paper will focus on a number of aspects of our studies of these porphyrins at different interfaces.The α, α, α, α (4,0) isomers are readily incorporated into Langmuir‐Blodgett films as either free base or metal complexes. Studies of assemblies containing free base and palladium (II) complexes have been carried out in which the porphyrin is irradiated in the presence of oxygen and nonexcited but oxidizable substrates. Much of the reactivity observed can be attributed to 1O2* generation. These studies reveal the migration range and reactivity of activated oxygen in a structure related to biomem‐branes.Several of these synthetic porphyrins have also been examined in cell suspensions and in synthetic reversed micelle solutions. Studies in the former have shown that the porphyrins can mediate the photoinactivation of several enzymes located inside and within the mitochondrial membrane in tumor cells extracted from rats. They are found to compare favorably to hematoporphyrin derivative in effectiveness.Studies of the same porphyrins in their reactivity towards copper ion incorporation in anionic reversed micelles indicate striking rate differences which can be interpreted in part to structural variations between the porphyrins as well as to their orientation at the hydrophobic‐hydrophilic interface.

Copper-albumin: What is its functional role?

The small copper fraction in animal blood that is bound to albumin protein is generally considered to have a transport role for the metal. However, several studies have concluded that copper ion incorporation into copper-enzymes requires caeruloplasmin to act as the transport form. The kinetic rates of Cu(albumin) reactions are also not in keeping with a general transport function. Only in the portal bloodstream does the Cu(albumin) appear to have a well-established transport role, i.e. in delivering the metal ions from the intestine to the liver. Thereafter the evidence as to its function is less certain; it could act as a storage form of the metal or have no role at all. Similar considerations apply to other metal-albumin fractions e.g. those of zinc and nickel.

The study of the gas—solid interaction of acetic acid with a cuprous oxide surface using reflection—absorption spectroscopy

The adsorption and reaction of acetic acid have been studied using the technique of reflection-absorption infrared spectroscopy. Evaporated copper films on both glass and gold substrates have been oxidized to cuprous oxide and allowed to interact with gaseous acetic acid. The results indicate that at ambient temperatures acetic acid adsorbs on cuprous oxide to form a vacuum stable (10 −7 Torr), thick (several hundred angstrom) film having an ionic lattice comprised primarily of discrete protons and acetate ions. At higher temperatures (near 200°C) incorporation of copper ions into the film occurs with apparent partial decomposition of the acetate ion. The general implications of these results on the understanding of the mechanisms of attack of organic acids on metal-metal oxide surfaces is discussed.

Glass processing

Copper(II) and nickel(II) complexes of sulfadimethoxine, sulfadiazine, sulfamerazine and sulfamethazine were synthesized with a good yield according to an original procedure. These complexes were first characterized by single-crystal X-ray diffraction and electrochemistry. Structural inspections showed that the antibacterial entity of ligands remains non-coordinated to metal ions in the complex high-lighting the fact that in each cluster, antiseptic activity of the metal has been associated to the antibiotic activity of the ligand. In order to confirm this possibility, antibacterial activities of the complexes were studied on several bacteria. The antibacterial activity of the complex is as important as the ligands one with the addition of antiseptic activity via the incorporation of copper ions.