Metal Ions For Catalytic Activity Research Articles

Biodegradable Nanoscale Coordination Polymers for Targeted Tumor Combination Therapy with Oxidative Stress Amplification

AbstractNowadays various inorganic nanoparticles that generate highly reactive hydroxyl radical (·OH) on the basis of Fenton‐like catalytic activity of metal ions have been designed for chemodynamic therapy. However, the high level of adaptive antioxidants [glutathione (GSH)] in cancer cells could effectively consume ·OH to compromise the treatment efficiency and biosafety of these inorganic nanoparticles, and this is a general concern in chemodynamic therapy. Herein, a new biodegradable nanoscale coordination polymer (NCP) is developed by integration of cisplatin prodrug (DSCP) and iron (III) ions through a reverse microemulsion method. The DSCP in the NCPs could react with GSH to release free cisplatin, while the iron (III) ions could be reduced by GSH into iron (II) to enable Fenton reaction, subsequently leading to amplified intracellular oxidative stress. After surface modification of polyethylene glycol (PEG) and cyclo[Arg‐Gly‐Asp‐D‐Phe‐Lys(mpa)] peptide (cRGD), Fe‐DSCP‐PEG‐cRGD shows an excellent targeting effect against αvβ3‐integrin overexpressed tumor cells. Furthermore, Fe‐DSCP‐PEG‐cRGD enables significant chemo and chemodynamic therapy with dramatically enhanced therapeutic efficiency in comparison to relative monotherapies. Importantly, Fe‐DSCP‐PEG‐cRGD could be efficiently cleared out from mice through feces and urine postinjection 7 days. The NCP presented in this work is simple and economical, which shows great biodegradability and biosafety for potential clinical translation.

Analysis of plant UDP-arabinopyranose mutase (UAM): Role of divalent metals and structure prediction

UDP-arabinopyranose mutase (UAM) is a plant enzyme which interconverts UDP-arabinopyranose (UDP-Arap; a six-membered sugar) to UDP-arabinofuranose (UDP-Araf; a five-membered sugar). Plant mutases belong to a small gene family called Reversibly Glycosylated Proteins (RGPs). So far, UAM has been identified in Oryza sativa (Rice), Arabidopsis thaliana and Hordeum vulgare (Barley). The enzyme requires divalent metal ions for catalytic activity. Here, the divalent metal ion dependency of UAMs from O. sativa (rice) and A. thaliana have been studied using HPLC-based kinetic assays. It was determined that UAM from these species had the highest relative activity in a range of 40–80μM Mn2+. Excess Mn2+ ion concentration decreased the enzyme activity. This trend was observed when other divalent metal ions were used to test activity. To gain a perspective of the role played by the metal ion in activity, an ab initio structural model was generated based on the UAM amino acid sequence and a potential metal binding region was identified. Based on our results, we propose that the probable role of the metal in UAM is stabilizing the diphosphate of the substrate, UDP-Arap.

In vitro selection of a sodium-specific DNAzyme and its application in intracellular sensing

Over the past two decades, enormous progress has been made in designing fluorescent sensors or probes for divalent metal ions. In contrast, the development of fluorescent sensors for monovalent metal ions, such as sodium (Na(+)), has remained underdeveloped, even though Na(+) is one the most abundant metal ions in biological systems and plays a critical role in many biological processes. Here, we report the in vitro selection of the first (to our knowledge) Na(+)-specific, RNA-cleaving deoxyribozyme (DNAzyme) with a fast catalytic rate [observed rate constant (ko(bs)) ∼ 0.1 min(-1)], and the transformation of this DNAzyme into a fluorescent sensor for Na(+) by labeling the enzyme strand with a quencher at the 3' end, and the DNA substrate strand with a fluorophore and a quencher at the 5' and 3' ends, respectively. The presence of Na(+) catalyzed cleavage of the substrate strand at an internal ribonucleotide adenosine (rA) site, resulting in release of the fluorophore from its quenchers and thus a significant increase in fluorescence signal. The sensor displays a remarkable selectivity (>10,000-fold) for Na(+) over competing metal ions and has a detection limit of 135 µM (3.1 ppm). Furthermore, we demonstrate that this DNAzyme-based sensor can readily enter cells with the aid of α-helical cationic polypeptides. Finally, by protecting the cleavage site of the Na(+)-specific DNAzyme with a photolabile o-nitrobenzyl group, we achieved controlled activation of the sensor after DNAzyme delivery into cells. Together, these results demonstrate that such a DNAzyme-based sensor provides a promising platform for detection and quantification of Na(+) in living cells.

Transition metal catalyzed mineralization of Calcon and bioassay of the mineralized solutions by Escherichia coli colony forming unit assay

Mineralization of Calcon was studied by Fenton and Fenton like processes in a wide range of pH with hydrogen peroxide (HP) and ammonium persulphate (APS) as oxidants and catalyzed by transition metal catalysts such as Fe2+, Ag+ and Co2+. Maximum mineralization for Fenton and other processes was achieved at pH 3 and 1, respectively. Although mineralization was negligible in alkaline media, the catalytic activity of metal ions on HP is found to be similar, while that on APS was in the order: Fe2+>Co2+>Ag+. The study reveals that Calcon resisted mineralization by Fenton process (35.6% in 120min) and the efficiency of other systems followed the order: Fe2+/APS>Ag+/APS≈Ag+/HP>Co2+/HP>Co2+/APS and a maximum of 91.1% and 73.7% mineralization were achieved with Fe2+/APS and Ag+/APS systems, respectively, at pH 1. To understand the impact of these processes operating at low pH, biotoxicity of the treated solutions was evaluated on the basis of Escherichia coli (E. coli) growth inhibition bioassay. It was observed that the treated solutions were still more toxic to E. coli than the original dye solution. Thus, metal catalyzed Fenton and Fenton like processes are not environmentally benign under the specified conditions of our study.

Interaction of mineral salts with the skin: a literature survey

There is growing scientific evidence that the health, well-being and the attractiveness of the skin are strongly influenced by nutrition. Consumers recognize this and have supported the creation of a global cosmeceuticals market estimated in 2010 at $27.2 billion. Early in 2011, the US Department of Health and Human Services and Department of Agriculture issued the Dietary Guidelines for Americans, 2010. Twelve vitamins and nine minerals were recognized as essential. The minerals include calcium, copper, iron, magnesium, phosphorus, selenium, zinc, potassium and sodium. Although the topical benefits of several minerals such as zinc, magnesium and iron are recognized and, in some cases, approved by the FDA, the topical benefits of the others to the skin are largely unexplored and unexploited. This review attempts to summarize what has been published in the literature on the interactions of the eight of the nine essential elements with the skin.

Bifidobacterium longum l-Arabinose Isomerase—Overexpression in Lactococcus lactis, Purification, and Characterization

Bifidobacterium longum NRRL B-41409 L-arabinose isomerase (L-AI) was cloned and overexpressed in Lactococcus lactis using a phosphate-depletion-inducible expression system. The purified B. longum L-AI was characterized using D-galactose and L-arabinose as the substrates. The enzyme was active and stable at acidic pH with an optimum at pH 6.0-6.5. The enzyme showed the highest activity at 55 °C during a 20-min incubation at pH 6.5. The K(m) value was 120 mM for L-arabinose and 590 mM for D-galactose. The V(max) was 42 U mg(-1) with L-arabinose and 7.7 U mg(-1) with D-galactose as the substrates. The enzyme had very low requirement for metal ions for catalytic activity, but it was stabilized by divalent metal ions (Mg(2+), Mn(2+)). The enzyme bound the metal ions so tightly that they could not be fully removed from the active site by EDTA treatment. Using purified B. longum L-AI as the catalyst at 35 °C, equilibrium yields of 36 % D-tagatose and 11 % L-ribulose with 1.67 M D-galactose and L-arabinose, respectively, as the substrates were reached.

Electronic and geometric structures of the organophosphate-degrading enzyme from Agrobacterium radiobacter (OpdA)

The organophosphate-degrading enzyme from Agrobacterium radiobacter (OpdA) is a highly efficient catalyst for the degradation of pesticides and some nerve agents such as sarin. OpdA requires two metal ions for catalytic activity, and hydrolysis is initiated by a nucleophilic hydroxide that is bound to one of these metal ions. The precise location of this nucleophile has been contentious, with both a terminal and a metal-ion-bridging hydroxide as likely candidates. Here, we employed magnetic circular dichroism to probe the electronic and geometric structures of the Co(II)-reconstituted dinuclear metal center in OpdA. In the resting state the metal ion in the more secluded αsite is five-coordinate, whereas the Co(II) in the solvent-exposed βsite is predominantly six-coordinate with two terminal water ligands. Addition of the slow substrate diethyl 4-methoxyphenyl phosphate does not affect the αsite greatly but lowers the coordination number of the βsite to five. A reduction in the exchange coupling constant indicates that substrate binding also triggers a shift of the μ-hydroxide into a pseudoterminal position in the coordination sphere of either the α or the β metal ion. Mechanistic implications of these observations are discussed.

Mechanistic Studies of Bacillus subtilis QueF, the Nitrile Oxidoreductase Involved in Queuosine Biosynthesis

The enzyme QueF was recently identified as an enzyme involved in the biosynthesis of queuosine, a 7-deazaguanosine modified nucleoside found in bacterial and eukaryotic tRNA. QueF exhibits sequence homology to the type I GTP cyclohydrolases characterized by FolE, but contrary to the predictions based on sequence analysis the enzyme in fact catalyzes a mechanistically unrelated reaction, the NADPH-dependent reduction of 7-cyano-7-deazaguanine (preQ0) to 7-aminomethyl-7-deazaguanine (preQ1), a late step in the queuosine pathway. The reduction of a nitrile is unprecedented in biology, and we report here characterization and mechanistic studies of the enzyme from Bacillus subtilis. The recombinant enzyme exhibits optimal activity at pH 7.5 and moderate ionic strength, and is not dependent on metal ions for catalytic activity. Steady-state kinetic analysis provided a kcat = 0.66 +/- 0.04 min-1, KM (preQ0) = 0.237 +/- 0.045 microM, and KM (NADPH) = 19.2 +/- 1.1 microM. Based on sequence analysis and homology modeling we predicted previously that Cys55 would be present in the active site and in proximity to the nitrile group of preQ0. Consistent with that prediction we observed that the enzyme was inactivated when preincubated with iodoacetamide, and protected from inactivation when preQ0 was present. Furthermore, titrations of the enzyme with preQ0 in the absence of NADPH were accompanied by the appearance of a new absorption band at 376 nm in the UV-vis spectrum consistent with the formation of an alpha,beta-unsaturated thioimide. Site-directed mutagenesis of Cys55 to Ala or Ser resulted in loss of catalytic activity and no absorption at 376 nm upon addition of preQ0. Based on our data we propose a chemical mechanism for the enzyme-catalyzed reaction, and a chemical rationale for the observation of covalent catalysis.

The activity of the dinuclear cobalt-β-lactamase from Bacillus cereus in catalysing the hydrolysis of β-lactams

Metallo-beta-lactamases are native zinc enzymes that catalyse the hydrolysis of beta-lactam antibiotics, but are also able to function with cobalt(II) and require one or two metal-ions for catalytic activity. The hydrolysis of cefoxitin, cephaloridine and benzylpenicillin catalysed by CoBcII (cobalt-substituted beta-lactamase from Bacillus cereus) has been studied at different pHs and metal-ion concentrations. An enzyme group of pK(a) 6.52+/-0.1 is found to be required in its deprotonated form for metal-ion binding and catalysis. The species that results from the loss of one cobalt ion from the enzyme has no significant catalytic activity and is thought to be the mononuclear CoBcII. It appears that dinuclear CoBcII is the active form of the enzyme necessary for turnover, while the mononuclear CoBcII is only involved in substrate binding. The cobalt-substituted enzyme is a more efficient catalyst than the native enzyme for the hydrolysis of some beta-lactam antibiotics suggesting that the role of the metal-ion is predominantly to provide the nucleophilic hydroxide, rather than to act as a Lewis acid to polarize the carbonyl group and stabilize the oxyanion tetrahedral intermediate.

Enzyme Deactivation Due to Metal-Ion Dissociation during Turnover of the Cobalt-β-Lactamase Catalyzed Hydrolysis of β-Lactams

Metallo-beta-lactamases are native zinc enzymes that catalyze the hydrolysis of beta-lactam antibiotics but are also able to function with cobalt (II) and require one or two metal ions for catalytic activity. The kinetics of the hydrolysis of benzylpenicillin catalyzed by cobalt substituted beta-lactamase from Bacillus cereus (BcII) are biphasic. The dependence of enzyme activity on pH and metal-ion concentration indicates that only the di-cobalt enzyme is catalytically active. A mono-cobalt enzyme species is formed during the catalytic cycle, which is virtually inactive and requires the association of another cobalt ion for turnover. Two intermediates with different metal to enzyme stoichiometries are formed on a branched reaction pathway. The di-cobalt enzyme intermediate is responsible for the direct catalytic route, which is pH-independent between 5.5 and 9.5 but is also able to slowly lose one bound cobalt ion via the branching route to give the mono-cobalt inactive enzyme intermediate. This inactivation pathway of metal-ion dissociation occurs by both an acid catalyzed and a pH-independent reaction, which is dependent on the presence of an enzyme residue of pK(a) = 8.9 +/- 0.1 in its protonated form and shows a large kinetic solvent isotope effect (H(2)O/D(2)O) of 5.2 +/- 0.5, indicative of a rate-limiting proton transfer. The pseudo first-order rate constant to regenerate the di-cobalt beta-lactamase from the mono-cobalt enzyme intermediate has a first-order dependence on cobalt-ion concentration in the pH range 5.5-9.5. The second-order rate constant for metal-ion association is dependent on two groups of pK(a) 6.32 +/- 0.1 and 7.47 +/- 0.1 being in their deprotonated basic forms and one group of pK(a) 9.48 +/- 0.1 being in its protonated form.

1.6 Å Crystal structure of a PA2721 protein from pseudomonas aeruginosa—A potential drug‐resistance protein

1.6 Å Crystal structure of a PA2721 protein from <i>pseudomonas aeruginosa</i>—A potential drug‐resistance protein

Inhibitory activity of thermal copolymers of amino acids for the metal-catalyzed hydrolysis of an RNA dinucleotide

It is well known that the hydrolysis of RNA is substantially catalyzed by several metal ions. Although this fact poses a problem for the RNA world hypothesis, there may have been unknown pathways for the protection of RNA molecules against the hydrolytic degradation under the primitive earth conditions. Thus, we have investigated whether or not thermal copolymers of amino acids (TCAA) inhibit the catalytic activity of metal ions for the RNA hydrolyses; TCAA is a suitable model material for prebiotic protein-like molecules since TCAA involving peptide bonding is readily prepared by heating amino acid mixtures under prebiotic conditions. The activities of metal ions that Fe(III) and Co(II) enhance somewhat the 3′,5′-cytidylylguanosine ( C 3 ′ pG ) hydrolysis and Ce(III) and Eu(III) accelerate greatly the C 3 ′ pG hydrolysis were notably reduced by TCAA. This fact indicates that protein-like molecules would have played important roles for the accumulation of RNA under the primitive earth conditions.

The Quorum-Quenching Lactonase from Bacillus thuringiensis Is a Metalloprotein

Lactonases from Bacillus species hydrolyze the N-acylhomoserine lactone (AHL) signaling molecules used in quorum-sensing pathways of many Gram-negative bacteria, including Pseudomonas aeruginosa and Erwinia carotovora, both significant pathogens. Because of sequence similarity, these AHL lactonases have been assigned to the metallo-beta-lactamase superfamily of proteins, which includes metalloenzymes of diverse activity, mechanism, and metal content. However, a recent study claims that AHL lactonase from Bacillus sp. 240B1 is not a metalloprotein [Wang, L. H., et al. (2004) J. Biol. Chem. 279, 13645]. Here, the gene for an AHL lactonase from Bacillus thuringiensis is cloned, and the protein is expressed, purified, and found to bind 2 equiv of zinc. The metal-bound form of AHL lactonase catalyzes the hydrolysis of N-hexanoyl-(S)-homoserine lactone but not the (R) enantiomer. Removal of both zinc ions results in loss of activity, and reconstitution with zinc restores activity, indicating the importance of metal ions for catalytic activity. Metal content, sequence alignments, and X-ray absorption spectroscopy of the zinc-containing lactonase all support a proposed dinuclear zinc binding site similar to that found in glyoxalase II.

Catalytic Activity of Metal Ions in Redox Processes in Polymer‐Salt Systems During Synthesis of Mixed Oxides

AbstractFor Abstract see ChemInform Abstract in Full Text.

Catalytic Activity of Metal Ions in Redox Processes in Polymer–Salt Systems during Synthesis of Mixed Oxides

Data are presented on the catalytic activity of metal ions in the oxidative destruction of poly(vinyl alcohol) in polymer–salt (nitrate) systems. The results indicate that the catalytic activity is due to the coexistence of oxidized and reduced forms of metals and that the oxidation potential for the conversion from one oxidation state to another must be high enough for catalysis. To assess catalytic activity, one can also compare the Gibbs energies of redox reactions involving oxides. Metal ions exhibit catalytic activity in solutions (of iron nitrate) and films (of molybdenum, tungsten, and silver salts) and also during the pyrolysis of polymer–salt systems in the synthesis of mixed oxides or in the presence of silver nitrate.

Purification and characterization of two isoapyrases from Solanum tuberosum var. ultimus

Two isoenzymes of ATP-diphosphohydrolase (apyrase) were extracted and purified from S. tuberosum var. Ultimus. Their hydrolytic activity ratios (ATPase/ADPase) were 1.0 (apyrase B) and ca 15.0 (apyrase A). They were characterized and compared with apyrases of other varieties of S. tuberosum. Ultimus apyrases, like the other apyrases, did not hydrolyse esteric bonds but only pyrophosphate bonds of organic and inorganic compounds. The optimum pH of all the studied hydrolytic activities of the Ultimus apyrases A and B was 6, except for the ADPase of enzyme A which was 8. Both enzymes require bivalent metal ions for catalytic activity. The activation order for both Ultimus enzymes was: Ca 2+>Mn 2+> Mg 2+>Co 2+>Zn 2+. Chemical modification of tryptophan, tyrosine, arginine and carboxylic residues decreased all enzymic activities of both apyrases. The modification of histidine residues reduced the ATPase and ADPase activities of the low ratio apyrase and the ATPase of the high ratio enzyme but did not affect its ADPase activity. Neither of the Ultimus apyrases showed the participation of -SH groups in the active site. The pI values obtained were: 5.45 for apyrase B and 6.56 for apyrase A. The absorption and the fluorescence spectra of the Ultimus isoenzymes were coincident. The amino acid composition of both isoenzymes is very similar, the number of histidines being the most remarkable difference. The amino acid composition of both isoenzymes does not explain the difference of one pH unit in the isoelectric point between the Ultimus enzymes A and B.

Dynamics of ternary complex formation and dissociation

Abstract

Catalytic effect of various metal ions on the methylation of mercury in the presence of humic substances

Methylation of Hg2+ (Hg(NO3)2) in the presence of fulvic acid (FA) and various metal ions has been studied. The concentrations of Hg2+ and FA ranged from 5 to 20 mg L−1 and 171 to 285 mg L−1 DOC, respectively. The pH range was 3 to 6.5. FA was isolated from an acid brown-water lake by XAD-8 polymeric adsorbent. Methylmercury production in the dark during 2 to 4 days incubation at 30 °C increased with increasing concentrations of Hg2+ ion and FA as well as with additions of metal ions (5 to 10 × 10−5 mole L−1 The observed catalytic activity of metal ions followed the order Fe3+ (Fe2+) > Cu2+ ≈ Mn2+, > Al3+. The production of methylmercury had a pH-optimum around 4 to 4.5 at the conditions tested.

Homogeneous catalytic oxidation of pyrocatechol in the presence of transition metal ions

Pyrocatechol oxidation by oxygen in aqueous solutions containing different metal ions, proceeds via the intermediate formation of pyrocatechol-metal ion-oxygen complexes, Catalytic activity of metal ions depends on their donor-acceptor properties.

Reduction by a Model of NAD(P)H. XIV. Mechanistic Consideration on the Role of Metal Ion

Abstract The driving force for the catalytic activity of metal ions on the reduction of α-keto esters with an NAD(P)H-model compound has been discussed. The scope of the reaction and spectroscopic investigations as well as molecular orbital consideration have revealed that the transition state of the reaction consists of a ternary complex in analogy with a coenzyme–enzye–substrate complex in an enzymic system. It is concluded that, at the transition state, one electron migrates from a model compound to a substrate through a metal ion, which is followed by the transfer of a proton.