Publications

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    ABSTRACT: The accumulation of plagues of β-amyloid (Aβ) peptides in the brain is a hallmark of Alzheimer's disease (AD). The redox-active Cu and Fe complexes of Aβ can cause damage to the neurons potentially via reactive oxygen species (ROS). The significant metal-mediated oxidative activity of CuAβ suggests that its presence can be chemically devastating regardless whether it is a cause or a result of AD. Flavonoids exhibit various benefits to human health, attributable to their metal-binding and antioxidation activities to certain extents. Despite broad interests and extensive studies of their metal-binding properties and anti/pro-oxidation activities, these properties and the mechanisms of the activities toward metal-centered oxidation reactions have not been fully revealed and concluded. We report herein distinctive antioxidation mechanisms between two flavonoid families toward the oxidation reactions by CuAβ(1-20), wherein the flavonols quercetin (Qr) and myricetin (Mr) competitively inhibit the oxidation of catechol by CuAβ(1-20) with K(i) of 11.2 and 32.6 μM, respectively, whereas the flavanols catechin (Ct) and epicatechin (Et) are substrates with k(cat) = 1.01 × 10(-2) and 1.55 × 10(-3) s(-1) and K(m) = 0.94 and 0.55 mM, respectively. Qr has a nearly 10-fold higher antioxidative efficacy than Ct against the oxidation activity of CuAβ, while Ct is effectively oxidized, which further decreases its antioxidant capacity. Similar inhibition patterns are observed toward oxidation of the catecholamine neurotransmitter dopamine by CuAβ(1-20). Metal ions and CuAβ bind Qr with a 1:1 ratio under our experimental conditions through the α-ketoenolate moiety as determined by the use of Co(2+) and Yb(3+) as paramagnetic NMR probes. Unlike flavanols, which are merely suicide antioxidative substrates, flavonols bind to the metal center and prevent metal-mediated redox reactions. We suggest flavonols may serve as leads for drug discovery and/or as agents toward preventing metal-mediated oxidative stress due to AD and other disorders. Moreover, CuAβ shows 8.6- and 4.2-fold higher kinetic regioselectivity in terms of k(cat) and k(cat)/K(m), respectively, toward the peroxidation of Ct than that of the enantiomer Et, suggesting potential development of metallo-catalysts in regioselective catalysis by the use of metallopeptides as templates.
    Inorganic Chemistry 01/2013; · 4.59 Impact Factor
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    ABSTRACT: The protein myoglobin has been successfully immobilized into a mesoporous metal-organic framework with hierarchical pore sizes, which demonstrates interesting size-selective biocatalysis as well as superior catalytic activities toward small substrate oxidation compared to its mesoporous silica material counterpart.
    Inorganic Chemistry 08/2012; 51(17):9156-8. · 4.59 Impact Factor
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    ABSTRACT: It has been demonstrated for the first time that the heme protein cytochrome c (Cyt c) can enter the interior of a MOF despite the larger molecular dimension of the protein relative to the access pore sizes. Mechanistic studies suggest that the Cyt c molecules must undergo a significant conformational change during translocation into the MOF interior through the relatively small nanopores.
    Journal of the American Chemical Society 08/2012; 134(32):13188-91. · 10.68 Impact Factor
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    ABSTRACT: Cu(II) complexes of Alzheimer's disease-related β-amyloid (Aβ) peptides exhibit metal-centered oxidation chemistry. The metallo-Aβ complexes are the hallmark of the disease and have been attributed to the generation of reactive oxygen species (ROS), causing oxidative stress. In this communication, the inhibitions of the oxidative activity of Cu(II)-Aβ by vitamin B6 compounds pyridoxamine (PM), pyridoxine (PN), pyridoxal (PL), and pyridoxal-5'-phosphate (PLP) are presented. These B6's are competitive inhibitors toward dopamine oxidation by Cu(II)-Aβ(1-20), with K(i) values of 1.4, 8.3, 1.2, and 0.2mM, respectively. The phospho-moiety in PLP seems to exhibit cooperative inhibition, affording a clue for future design of inhibitors.
    Bioorganic & medicinal chemistry letters 09/2011; 21(21):6430-2. · 2.65 Impact Factor
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    ABSTRACT: Microperoxidase-11 has for the first time been successfully immobilized into a mesoporous metal-organic framework (MOF) consisting of nanoscopic cages and it demonstrates superior enzymatic catalysis performances compared to its mesoporous silica counterpart.
    Journal of the American Chemical Society 06/2011; 133(27):10382-5. · 10.68 Impact Factor
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    ABSTRACT: A multidentate imidazole-containing cyclophosphazene (Im6Cpz) ligand was synthesized and its metal complexes (MxIm6Cpz; M = ZnII, CuII, CoII; x = 1, 2, 3) were prepared and used as phosphoesterase models towards hydrolysis of the model substrates p-nitrophenylphosphate (NPP) and bis(p-nitrophenyl)phosphate (BNPP) in 75 % DMSO buffer solution at pH = 7–11 at 37 °C. The hydrolysis of BNPP by Cu3Im6Cpz exhibits enzyme-like saturation kinetics with kcat = 1.4 × 10–5 s–1 and kcat/Km = 0.0027 M–1 s–1. The CuII complex exhibits a tremendous selectivity toward NPP, showingnearly a stoichiometric binding with kcat = 6.8 × 10–4 s–1 under saturation conditions and a significant second-order rate constant kcat/Kd = 136 M–1 s–1, which is 5.0 × 104 times higher than that for BNPP hydrolysis. An intramolecular dinuclear pathway was revealed for the complexes Cu2Im6Cpz andCu3Im6Cpz; whereas an intermolecular dinuclear pathway was observed for CuIm6Cpz. Cu3Im6Cpz shows an order of magnitude higher activity than the simple CuII complex of the untethered ligand N-methylimidazole, Cu(MeIm)2, suggesting a possible contribution from the proximity effect in the Im6Cpz complexes. The high catalytic specificity of Cu3Im6Cpz towards the phosphomonoester NPP suggests that it can serve as a good model system and a blueprint for further exploration of catalytic activity and specificity in phosphoester hydrolysis.
    Berichte der deutschen chemischen Gesellschaft 01/2011; 2011(5):674 - 682. · 2.94 Impact Factor
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    ABSTRACT: High-energy irradiations of polymers may cause bond cleavage or cross-linking and change the structures and physical properties of the polymers, which may offer various applications. Despite wide investigation, relationship between the kinetics and mechanism of annihilation after irradiations and the structure and some physical properties of irradiated polymers is still poorly established. We have been exploring such possible relationship and report herein investigation of the kinetics of radical annihilation of γ-ray irradiated acrylic, i.e., poly(methylmethacrylate) or PMMA, at elevated temperatures with EPR spectroscopy. The EPR spectra consist of three components, a quintet Ra, a quartet Rb, and a broad singlet Rc. Ra and Rb follow second-order annihilation kinetics, while the decay of the radical Rc is comprised of at least two parallel kinetic processes, a slow second-order pathway and a fast pathway which can be equally well fitted to first- or second-order kinetics. The kinetics is analogous to that for the radical decays in irradiated 2-hydroxyethyl methacrylate copolymer. On the basis of the large hyperfine coupling constant of 230 mT, Ra may be assigned to a radical adjacent to two groups of protons, such as a doublet of quartet with similar coupling constants due to an anti-methylene proton and a methyl group; the Rb signal, possibly a methyl radical; and the broad singlet Rc, a magnetically coupled combination species. Alternative assignments of the radicals have also been suggested. The rate constant increases with increasing dose for each radical at a given temperature, possibly due to increase in radical concentrations at higher doses. The rate constants satisfy the Arrhenius equation, suggesting a single mechanistic pathway for the annihilation process in the temperature range; wherein the activation energy decreases with increasing dose for all radicals, possibly due to higher concentrations of free radicals in close proximity produced at higher doses.
    Polymer. 01/2011;
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    ABSTRACT: The annihilation of the radicals in irradiated 2-hydroxyethyl methacrylate copolymer was analyzed by the use of electron paramagnetic resonance (EPR) spectroscopy. The EPR spectra were deconvoluted into three radicals: a quartet (Ra), a triplet (Rb), and a broad singlet (Rc). Radical Ra was attributed to coupling with a methyl radical and/or a doublet or triplet with about the same hyperfine coupling due to a methylene radical. Radical Rb was due to a methylene radical produced by main-chain scission. Radical Rc was attributed to various free radicals without coupling to protons. By comparing the EPR spectra of radicals Ra, Rb, and Rc with the spectrum of a 2,2-diphenyl-1-picrylhydrazyl (DPPH) standard with a known spin number, we calculated the spin numbers of the radicals, which decreased with time in the temperature range 25–45°C, regardless of the irradiation dose. The annealing of Ra and Rb and the annealing of Rc at longer times followed second-order kinetics; these were different from the kinetics for the color formation and defect-controlled hardening of polymers. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010
    Journal of Applied Polymer Science 09/2010; 117(6):3114 - 3120. · 1.40 Impact Factor
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    ABSTRACT: The peptidyl antibiotic bacitracin (Bc) is one of the most widely used antibiotics which can bind divalent transition metal ions, including Mn(II), Co(II), Ni(II), Cu(II), and Zn(II). The metal binding is essential for its antimicrobial activity. Previous analysis of the hyperfine-shifted (1)H NMR signals of Co(II)-Bc A(1) revealed the structure of the metal binding environment and a potential hydrophobic site important for the bioactivity of this antibiotic. Co(II)-Bc in DMSO shows relatively sharper hyperfine-shifted (1)H NMR signals compared with the spectrum acquired in an aqueous solution, allowing more thorough analysis of the signals with 1D and 2D NMR methods. Pyrophosphate and derivatives bind to Co(II)-Bc to form kinetically inert ternary complexes. The coordinated D-Glu-4 is found detached from the metal center of metallobacitracin upon trimetaphosphate binding, implying its role in the antibiotic activity of Bc. We further demonstrate in this report the structure-function relationship on desamido-Bc of low antibiotic activity by the use of NMR, wherein D-Glu-4 is suggested to be important for the bioactivity of Bc. The interaction of the phospho-moiety with Bc is also reflected by DNA binding, wherein metal-free Bc does not bind DNA, whereas various metal complexes of Bc do. Cu(II)-Bc was further demonstrated to bind and oxidatively cleave DNA under reduction conditions in the air. It also exhibited a significant oxidative activity toward catechol oxidation, showing enzyme-like saturation kinetics with k(cat) = 7.0 x 10(-3) s(-1) and k(cat)/K(m) = 2.1 M(-1) s(-1) aerobically and k(cat) = 0.38 s(-1) and k(cat)/K(m) = 14.7 M(-1) s(-1) in the presence of 32 mM of H(2)O(2). The binding of pyrophosphate moiety to metallobacitracin, the detachment of d-Glu-4, and the significant oxidative activity of Cu(II)-Bc provide further insights into the bioactivity of this metallopeptide and Cu-oxygen chemistry.
    Journal of the American Chemical Society 04/2010; 132(16):5652-61. · 10.68 Impact Factor
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    ABSTRACT: Histatin-5 (Hn5) is an antimicrobial salivary peptide of 24 amino acids. Two specific metal-binding sites were revealed with electronic, NMR, and EPR spectroscopy. The complex Cu(2)(II)-Hn5 effectively oxidizes catechol, exhibiting enzyme-like kinetics (k(cat)=0.011 and 0.060 s(-1) and k(cat)/K(m)=19 and 50 M(-1)s(-1) without and with 12.8mM H(2)O(2), respectively). The significant oxidative activity may contribute to the biological activity of this antibiotic metallopeptide.
    Bioorganic & medicinal chemistry letters 12/2009; 19(23):6709-12. · 2.65 Impact Factor
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    ABSTRACT: The dinuclear aminopeptidase from Streptomyces griseus (SgAP) and its metal derivatives catalyze the hydrolysis of the phosphoester bis(p-nitrophenyl) phosphate (BNPP) and the phosphonate ester p-nitrophenyl phenylphosphonate with extraordinary rate enhancements at pH 7.0 and 25 degrees C [A. Ercan, H. I. Park, L.-J. Ming, Biochemistry 45, (2006) 13779-13793.], reaching 6.7 billion-fold in terms of the first-order rate constant of the di-Co(II) derivative with respect to the autohydrolytic rates. Since phosphoesters are transition state-like inhibitors in peptide hydrolysis, their hydrolysis by SgAP is quite novel. Herein, we report the investigation of this proficient alternative catalysis of SgAP and the role of each metal ion in the dinuclear site toward peptide and BNPP hydrolysis. Mn(II) selectively binds to one of the dinuclear metal sites (M1), affording MnE-SgAP with an empty (E) second site for the binding of another metal (M2), including Mn(II), Co(II), Ni(II), Zn(II), and Cd(II). Peptide hydrolysis is controlled by M2, wherein the k(cat) values for the derivatives MnM2-SgAP are different yet similar between MnCo- and CoCo-SgAP and pairs of other metal derivatives. On the other hand, BNPP hydrolysis is affected by metals in both sites. Thus, the two hydrolytic catalyses must follow different mechanisms. Based on crystal structures, docking, and the results presented herein, the M1 site is close to the hydrophobic specific site and the M2 site is next to Tyr246 that is H-bonded to a coordinated nucleophilic water molecule in peptide hydrolysis; whereas a coordinated water molecule on M1 becomes available as the nucleophile in phosphodiester hydrolysis.
    Journal of inorganic biochemistry 09/2009; 104(1):19-29. · 3.25 Impact Factor
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    ABSTRACT: Fe3+ complexes of pyridine-containing copolymers were found to be efficient and selective catalysts toward phosphodiester hydrolysis and show significant activity toward oxidative DNA cleavage. The catalysis toward bis(p-nitrophenyl)phosphate (BNPP) hydrolysis exhibits enzyme-like pre-equilibrium kinetics with maximum activities in the range of ca. pH 6–8 and a first-order catalytic proficiency (kcat/ko) of 4.2 × 107-fold at the acidic pH value of 5.3 (i.e., pKa of the coordinated nucleophilic water) and 25 °C, entitling this Fe3+ copolymer an acid phosphodiester catalyst. This catalyst also shows significant selectivity toward BNPP hydrolysis relative to the hydrolyses of p-nitrophenyl phenylphosphonate and p-nitrophenylphosphate monoester, with a ratio of 4250:16:1 in terms of their first-order catalytic proficiencies at pH 8.0 and 25 °C. Fe3+ complexes of a few pyridine-containing copolymers show different hydrolytic activities, which points a direction for rational design of catalytic metallopolymers. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
    Berichte der deutschen chemischen Gesellschaft 02/2009; 2009(9):1199 - 1207. · 2.94 Impact Factor
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    ABSTRACT: The potential risk of metal-centered oxidative catalysis has been overlooked in the research of the copper complexes of the Alzheimer's disease-related beta-amyloid (Abeta) peptides. Cu(2+) complexes of Abeta(1-40) and its 1-16 and 1-20 fragments have recently been shown to exhibit significant metal-centered oxidative activities toward several catecholamine neurotransmitters with and without H(2)O(2) around neutral pH [G.F.Z. da Silva, L.-J. Ming, "Metallo-ROS" in Alzheimer's disease: metal-centered oxidation of neurotransmitters by Cu(II)-beta-amyloid and neuropathology of Alzheimer's disease, Angew. Chem. Int. Ed. 46 (2007) 3337-3341]. The results further support the metallo-Abeta-associated oxidative stress theory often considered to be connected to the neuropathology of the disease. The metal-centered oxidative catalysis of CuAbeta(1-16/20) challenges the long-standing proposed redox role of Met35 in Abeta because Abeta(1-16/20) do not contain a Met. External Met has been determined by kinetic, optical, and electron paramagnetic resonance methods to bind directly to the Cu(2+) center of CuAbeta(1-40) and CuAbeta(1-20) with K(d)=2.8 mM and 11.3 microM, respectively, which reflects less accessibility of the metal center in the full-length CuAbeta(1-40). However, Met does not serve as a reducing agent for the Cu(II) which thus must amplify the observed oxidative catalysis of CuAbeta(1-20)through a non-redox mechanism. Conversely, the CuAbeta-catalyzed oxidation reaction of dopamine is inhibited by bio-available reducing agents such as ascorbate (competitive K(ic)=66 microM) and glutathione (non-competitive, K(inc)=53 microM). These data indicate that the oxidation chemistry of metallo-Abeta is not initiated by Met35. The results yield further molecular and mechanistic insights into the roles of metallo-Abeta in this disease.
    Biochimica et Biophysica Acta 12/2008; 1792(1):49-55. · 4.66 Impact Factor
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    ABSTRACT: A lipase-producing bacterium was isolated and identified as Pseudomonas monteilii TKU009. A lipase (F2) and lipase-like materials (F1) were purified from the culture supernatant of P. monteilii TKU009 with soybean powder as the sole carbon/nitrogen source. The molecular mass of F1 and F2 was estimated to be 44 kDa by SDS-PAGE and gel filtration. The optimum pH, optimum temperature, and pH and thermal stabilities of F2 were 7, 40 degrees C, 8-11, and 50 degrees C; and of F1 were 6, 40 degrees C, 6-7, and 50 degrees C, respectively. F2 was completely inhibited by EDTA and slightly by Mg(2+), Fe(2+), Mn(2+), and SDS. F1 was completely inhibited by EDTA and Fe(2+) and strongly by Zn(2+), Mn(2+), Ca(2+), Mg(2+), and SDS. The activities of both the enzymes were enhanced by the addition of non-ionic surfactants Triton X-100 and Tween 40, especially for F1. F2 preferably acted on substrates with a long chain (C10-C18) of fatty acids, while F1 showed a broad spectrum on those with chain length of C4-C18. The marked activity of F2 in organic solvents makes it an ideal choice for application in a water-restricted medium including organic synthesis.
    Journal of Industrial Microbiology 10/2008; 36(1):65-73. · 1.80 Impact Factor
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    ABSTRACT: amide copolymer exhibits very efficient 2-electron catalysis toward the oxidation of catechol and derivatives to form quin- ones with and without 80 mM (0.27%) H2O2, showing re- markable (0.114-2.67)10 5 and (2.83-9.60)10 4 -fold rate enhancements, respectively, in terms of first-order rate con- stant relative to auto-oxidation of the substrates in an aque- ous environment under mild conditions. Metal-binding pro- files suggest the presence of cooperativity in the catalysis. The oxidation catalysis is inhibited by the di-copper tyrosi-
    Berichte der deutschen chemischen Gesellschaft 04/2008; 2008(16):2584 - 2592. · 2.94 Impact Factor
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    ABSTRACT: Free radicals are formed upon irradiation of polymers. The annealing of γ-ray irradiated syndiotactic polystyrene (sPS) with doses 10, 23 and 36kGy was studied with electron paramagnetic resonance (EPR) spectroscopy. The EPR spectra are possibly attributed to three types of radicals, (a) the benzyl radical Ra, (b) resonance structure of the phenyl radical Rb, and (c) carbon-superoxide-centered radical Rc with three-, four-, and single-line features, respectively. Radical Ra can be easily formed by the removal of the proton from the tertiary carbon; radical Rb is created from delocalization of the spin of the radical Ra onto the benzene ring; and radical Rc may be generated from the interaction of a carbon-centered radical with dioxygen from the air which forms a carbon-superoxide center. By comparing EPR spectra of the radicals with a DPPH standard, the spin numbers of the radicals can be calculated. The spin number of all radicals decreases exponentially with time in the temperature range of 60–90°C regardless of dose of the irradiation. The annealing of Ra, Rb, and Rc follows first-order kinetics. The activation energies of the annihilation are determined to be 15.8–19.0, 16.0–19.5, and 23.2–26.6kJ/mol for radicals Ra, Rb, and Rc, respectively. The kinetic study presented herein can serve as a criterion for γ-ray irradiation process in various applications, such as sterilization of polymer materials and devices.
    Polymer. 01/2008; 49(18):3987-3992.
  • Giordano F Z da Silva, Li-June Ming
    Angewandte Chemie International Edition 02/2007; 46(18):3337-41. · 11.34 Impact Factor
  • Li-June Ming
    Angewandte Chemie. 01/2007; 119(18):3401-3405.
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    Altan Ercan, Hyun Ik Park, Li-June Ming
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    ABSTRACT: A unique "enzyme catalytic promiscuity" has recently been observed, wherein a phosphodiester and a phosphonate ester are hydrolyzed by a dinuclear aminopeptidase and its metal derivatives from Streptomyces griseus (SgAP) [Park, H. I., Ming, L.-J. (1999) Angew. Chem., Int. Ed. Engl. 38, 2914-2916 and Ercan, A., Park, H. I., Ming, L.-J. (2000) Chem. Commun. 2501-2502]. Because tetrahedral phosphocenters often serve as transition-state inhibitors toward the hydrolysis of the peptide, phosphoester hydrolysis by peptidases is thus not expected to occur effectively and must take place through a unique mechanism. Owing to the very different structures and mechanistic requirements between phosphoesters and peptides during hydrolysis, the study of this effective phosphodiester hydrolysis by SgAP may provide further insight into the action of this enzyme that is otherwise not obtainable from regular peptide substrates. We present herein a detailed investigation of both peptide and phosphodiester hydrolyses catalyzed by SgAP. The latter exhibits a first-order rate enhancement of 4 x 10(10)-fold compared to the uncatalyzed reaction at pH 7.0 and 25 degrees C. The results suggest that peptide and phosphodiester hydrolyses by SgAP may share a common reaction mechanism to a certain extent. However, their differences in pH dependence, phosphate and fluoride inhibition patterns, and proton inventory reflect that they must follow different pathways. Mechanisms for the two hydrolyses are drawn on the basis of the results, which provide the foundation for further investigation of the catalytic promiscuity of this enzyme by means of physical and molecular biology methods. The catalytic versatility of SgAP suggests that this enzyme may serve as a unique "natural model system" for further investigation of dinuclear hydrolysis. A better understanding of enzyme catalytic promiscuity is also expected to shed light on the evolution and action of enzymes.
    Biochemistry 12/2006; 45(46):13779-93. · 3.38 Impact Factor
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    ABSTRACT: Blastula protease 10 (BP10) is a metalloenzyme involved in sea urchin embryogenesis, which has been assigned to the astacin family of zinc-dependent endopeptidases. It shows greatest homology with the mammalian tolloid-like genes and contains conserved structural motifs consistent with astacin, tolloid, and bone morphogenetic protein 1. Astacin, a crustacean digestive enzyme, has been proposed to carry out hydrolysis via a metal-centered mechanism that involves a metal-coordinated "tyrosine switch." It has not been determined if the more structurally complex members of this family involved in eukaryotic development share this mechanism. The recombinant BP10 has been overexpressed in Escherichia coli, its metalloenzyme nature has been confirmed, and its catalytic properties have been characterized through kinetic studies. BP10 shows significant hydrolysis toward gelatin both in its native zinc-containing form and copper derivative. The copper derivative of BP10 shows a remarkable 960% rate acceleration toward the hydrolysis of the synthetic substrate N-benzoyl-arginine-p-nitroanilide when compared with the zinc form. The enzyme also shows calcium-dependent activation. These are the first thorough mechanistic studies reported on BP10 as a representative of the more structurally complex members of astacin-type enzymes in deuterostomes, which can add supporting data to corroborate the metal-centered mechanism proposed for astacin and the role of the coordinated Tyr. We have demonstrated the first mechanistic study of a tolloid-related metalloenzyme involved in sea urchin embryogenesis.
    Journal of Biological Chemistry 05/2006; 281(16):10737-44. · 4.65 Impact Factor

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