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ABSTRACT: Two new dinucleating ligands (H(3)L(2) and HL(3)), derivatives of a well-known dinucleating ligand (HL(1)) with two bis-picolylamine sites connected to a bridging phenolate, with hydrogen-bonding donor groups at two of the pyridine moieties were designed and synthesized. Design of these ligands suggests that they will lead to dinuclear complexes with potential to stabilize phosphoester substrates as monodentate rather than bridging ligands. We report the diferric complexes [Fe(III)(2)(H(2)L(2))(OH)](4+) and [Fe(III)(2)(L(3))(OH)(OH(2))(2)](4+), which have been characterized by spectrophotometric titrations, UV-vis, IR, NMR, EPR, and Mössbauer spectroscopy. The phosphatase activity of the diferric systems, in addition to the partially reduced heterovalent [Fe(III)Fe(II)(L(3))(OH)(OH(2))(2)](3+) complex, has been investigated, and the complexes are shown to catalytically hydrolyze the activated phosphodiester substrate BDNPP (bis-dinitrophenylphosphate) as well as the corresponding phosphomonoester substrate DNPP (dinitrophenylphosphate). The results indicate that indeed the secondary interactions lead to an increase of the phosphatase activity and to active phosphomonoesterase catalysts. Interestingly, the heterovalent form of the HL(3)-based complex is more efficient than the diferric complex, and this is also discussed.
Inorganic Chemistry 10/2012; · 4.60 Impact Factor
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ABSTRACT: A possible biological function of cyclic pseudo-octapeptides is presented. The dinuclear copper(II) complex of a synthetic analogue ([Cu(2)(H(2)Pat(1))(μ-OH)(OH(2))(2)]) of the naturally occurring ascidiacyclamide is known to have a hydroxo-bridged dicopper(II) site which is able to catalytically transform CO(2) into CO(3)(2-). This complex is shown here to function as a phosphatase mimic, suggesting that the so far unknown biological function of these macrocycles within the ascidians may involve phosphoester hydrolysis.
Chemical Communications 08/2012; 48(75):9364-6. · 6.17 Impact Factor
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ABSTRACT: Two synthetic derivatives of the naturally occurring cyclic pseudooctapeptides patellamide A-F and ascidiacyclamide, that is, H(4)pat(2), H(4)pat(3), as well as their Cu(II) complexes are described. These cyclic peptide derivatives differ from the naturally occurring macrocycles by the variation of the incorporated heterocyclic donor groups and the configuration of the amino acids connecting the heterocycles. The exchange of the oxazoline and thiazole groups by dimethylimidazoles or methyloxazoles leads to more rigid macrocycles, and the changes in the configuration of the side chains leads to significant differences in the folding of the cyclic peptides. These variations allow a detailed study of the various possible structural changes on the chemistry of the Cu(II) complexes formed. The coordination of Cu(II) with these macrocyclic species was monitored by high-resolution electrospray mass spectrometry (ESI-MS), spectrophotometric (UV/Vis) and circular dichroic (CD) titrations, and electron paramagnetic resonance (EPR) spectroscopy. Density functional theory (DFT) calculations and molecular mechanics (MM) simulations have been used to model the structures of the Cu(II) complexes and provide a detailed understanding of their geometric preferences and conformational flexibility. This is related to the Cu(II) coordination chemistry and the reactivity of the dinuclear Cu(II) complexes towards CO(2) fixation. The variation observed between the natural and various synthetic peptide systems enables conclusions about structure-reactivity correlations, and our results also provide information on why nature might have chosen oxazolines and thiazoles as incorporated heterocycles.
Chemistry 02/2012; 18(9):2578-90. · 5.93 Impact Factor
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Sarah J Smith,
Rosely A Peralta,
Rafael Jovito,
Adolfo Horn,
Adailton J Bortoluzzi,
Christopher J Noble, Graeme R Hanson,
Robert Stranger,
Vidura Jayaratne,
Germán Cavigliasso,
Lawrence R Gahan,
Gerhard Schenk,
Otaciro R Nascimento,
Angélica Cavalett,
Tiago Bortolotto,
Guilherme Razzera,
Hernán Terenzi,
Ademir Neves,
Mark J Riley
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ABSTRACT: A mixed-valence complex, [Fe(III)Fe(II)L1(μ-OAc)(2)]BF(4)·H(2)O, where the ligand H(2)L1 = 2-{[[3-[((bis(pyridin-2-ylmethyl)amino)methyl)-2-hydroxy-5-methylbenzyl](pyridin-2-ylmethyl)amino]methyl]phenol}, has been studied with a range of techniques, and, where possible, its properties have been compared to those of the corresponding enzyme system purple acid phosphatase. The Fe(III)Fe(II) and Fe(III)(2) oxidized species were studied spectroelectrochemically. The temperature-dependent population of the S = 3/2 spin states of the heterovalent system, observed using magnetic circular dichroism, confirmed that the dinuclear center is weakly antiferromagnetically coupled (H = -2JS(1)·S(2), where J = -5.6 cm(-1)) in a frozen solution. The ligand-to-metal charge-transfer transitions are correlated with density functional theory calculations. The Fe(III)Fe(II) complex is electron paramagnetic resonance (EPR)-silent, except at very low temperatures (<2 K), because of the broadening caused by the exchange coupling and zero-field-splitting parameters being of comparable magnitude and rapid spin-lattice relaxation. However, a phosphate-bound Fe(III)(2) complex showed an EPR spectrum due to population of the S(tot) = 3 state (J= -3.5 cm(-1)). The phosphatase activity of the Fe(III)Fe(II) complex in hydrolysis of bis(2,4-dinitrophenyl)phosphate (k(cat.) = 1.88 × 10(-3) s(-1); K(m) = 4.63 × 10(-3) mol L(-1)) is similar to that of other bimetallic heterovalent complexes with the same ligand. Analysis of the kinetic data supports a mechanism where the initiating nucleophile in the phosphatase reaction is a hydroxide, terminally bound to Fe(III). It is interesting to note that aqueous solutions of [Fe(III)Fe(II)L1(μ-OAc)(2)](+) are also capable of protein cleavage, at mild temperature and pH conditions, thus further expanding the scope of this complex's catalytic promiscuity.
Inorganic Chemistry 02/2012; 51(4):2065-78. · 4.60 Impact Factor
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ABSTRACT: The synthesis and characterization of a novel dinucleating ligand L (L=4,11-dimethyl-1,8-bis{2-[N-(di-2-pyridylmethyl)amino]ethyl}cyclam) and its μ-oxo-bridged diferric complex [(H(2)L){Fe(III)(2)(O)}(Cl)(4)](2+) are reported. This diiron(III) complex is the first example of a truly functional purple acid phosphatase (PAP) mimic as it accelerates the hydrolysis of the activated phosphomonoester 2,4-dinitrophenyl phosphate (DNPP). The spectroscopic and kinetic data indicate that only substrates that are monodentately bound to one of the two ferric ions can be attacked by a suitable nucleophile. This is, most probably, a terminal iron(III)-bound hydroxide. DFT calculations support this assumption and also highlight the importance of secondary interactions, exerted by the protonated cyclam platform, for the positioning and activation of the iron(III)-bound substrate. Similar effects are postulated in the native enzyme but addressed in PAP mimics for the first time.
Chemistry 02/2012; 18(6):1700-10. · 5.93 Impact Factor
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Sarah J. Smith,
Rosely A. Peralta,
Rafael Jovito,
Adolfo Horn,
Adailton J. Bortoluzzi,
Christopher J. Noble, Graeme R. Hanson,
Robert Stranger,
Vidura Jayaratne,
Germán Cavigliasso,
Lawrence R. Gahan,
Gerhard Schenk,
Otaciro R. Nascimento,
Angélica Cavalett,
Tiago Bortolotto,
Guilherme Razzera,
Hernán Terenzi,
Ademir Neves,
Mark J. Riley
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ABSTRACT: A mixed-valence complex, [FeIIIFeIIL1(μ-OAc)2]BF4·H2O, where the ligand H2L1 = 2-{[[3-[((bis(pyridin-2-ylmethyl)amino)methyl)-2-hydroxy-5-methylbenzyl](pyridin-2-ylmethyl)amino]methyl]phenol}, has been studied with a range of techniques, and, where possible, its properties have been compared to those of the corresponding enzyme system purple acid phosphatase. The FeIIIFeII and FeIII2 oxidized species were studied spectroelectrochemically. The temperature-dependent population of the S = 3/2 spin states of the heterovalent system, observed using magnetic circular dichroism, confirmed that the dinuclear center is weakly antiferromagnetically coupled (H = −2JS1·S2, where J = −5.6 cm–1) in a frozen solution. The ligand-to-metal charge-transfer transitions are correlated with density functional theory calculations. The FeIIIFeII complex is electron paramagnetic resonance (EPR)-silent, except at very low temperatures (<2 K), because of the broadening caused by the exchange coupling and zero-field-splitting parameters being of comparable magnitude and rapid spin–lattice relaxation. However, a phosphate-bound FeIII2 complex showed an EPR spectrum due to population of the Stot = 3 state (J= −3.5 cm–1). The phosphatase activity of the FeIIIFeII complex in hydrolysis of bis(2,4-dinitrophenyl)phosphate (kcat. = 1.88 × 10–3 s–1; Km = 4.63 × 10–3 mol L–1) is similar to that of other bimetallic heterovalent complexes with the same ligand. Analysis of the kinetic data supports a mechanism where the initiating nucleophile in the phosphatase reaction is a hydroxide, terminally bound to FeIII. It is interesting to note that aqueous solutions of [FeIIIFeIIL1(μ-OAc)2]+ are also capable of protein cleavage, at mild temperature and pH conditions, thus further expanding the scope of this complex’s catalytic promiscuity.
01/2012;
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ABSTRACT: Cytochrome P450 enzymes (P450s) are exceptionally versatile monooxygenases, mediating hydroxylations of unactivated C-H bonds, epoxidations, dealkylations, and N- and S-oxidations as well as other less common reactions. In the conventional view of the catalytic cycle, based upon studies of P450s in vitro, substrate binding to the Fe(III) resting state facilitates the first 1-electron reduction of the heme. However, the resting state of P450s in vivo has not been examined. In the present study, whole cell difference spectroscopy of bacterial (CYP101A1 and CYP176A1, i.e. P450cam and P450cin) and mammalian (CYP1A2, CYP2C9, CYP2A6, CYP2C19, and CYP3A4) P450s expressed within intact Escherichia coli revealed that both Fe(III) and Fe(II) forms of the enzyme are present in the absence of substrates. The relevance of this finding was supported by similar observations of Fe(II) P450 heme in intact rat hepatocytes. Electron paramagnetic resonance (EPR) spectroscopy of the bacterial forms in intact cells showed that a proportion of the P450 in cells was in an EPR-silent form in the native state consistent with the presence of Fe(II) P450. Coexpression of suitable cognate electron donors increased the degree of endogenous reduction to over 80%. A significant proportion of intracellular P450 remained in the Fe(II) form after vigorous aeration of cells. The addition of substrates increased the proportion of Fe(II) heme, suggesting a kinetic gate to heme reduction in the absence of substrate. In summary, these observations suggest that the resting state of P450s should be regarded as a mixture of Fe(III) and Fe(II) forms in both aerobic and oxygen-limited conditions.
Journal of Biological Chemistry 11/2011; 286(47):40750-9. · 4.77 Impact Factor
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ABSTRACT: Cytochrome P450 enzymes (P450s) are exceptionally versatile monooxygenases, mediating hydroxylations of unactivated C-H bonds,
epoxidations, dealkylations, N- and S-oxidations as well as other less common reactions. In the conventional view of the catalytic
cycle, based upon studies of P450s in vitro, substrate binding to the Fe(III) resting state facilitates the first one electron
reduction of the heme. However the resting state of P450s in vivo has not been examined. In the present study, whole cell
difference spectroscopy of bacterial (CYP101A1 and CYP176A1 i.e. P450cam and P450cin) and mammalian (CYP1A2, CYP2C9, CYP2A6,
CYP2C19 and CYP3A4) P450s expressed within intact Escherichia coli revealed that both Fe(III) and Fe(II) forms of the enzyme
are present in the absence of substrates. The relevance of this finding was supported by similar observations of Fe(II) P450
heme in intact rat hepatocytes. Electron paramagnetic resonance (EPR) spectroscopy of the bacterial forms in intact cells
showed that a proportion of the P450 in cells was in an EPR silent form in the native state consistent with the presence of
Fe(II) P450. Coexpression of suitable cognate electron donors increased the degree of endogenous reduction to over 80%. A
significant proportion of intracellular P450 remained in the Fe(II) form after vigorous aeration of cells. Addition of substrates
increased the proportion of Fe(II) heme suggesting a kinetic gate to heme reduction in the absence of substrate. In summary,
these observations suggest that the resting state of P450s should be regarded as a mixture of Fe(III) and Fe(II) forms in
both aerobic and oxygen-limited conditions.
Journal of Biological Chemistry 10/2011; · 4.77 Impact Factor
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ABSTRACT: SoxAX cytochromes catalyze the formation of heterodisulfide bonds between inorganic sulfur compounds and a carrier protein,
SoxYZ. They contain unusual His/Cys-ligated heme groups with complex spectroscopic signatures. The heme-ligating cysteine
has been implicated in SoxAX catalysis, but neither the SoxAX spectroscopic properties nor its catalysis are fully understood
at present. We have solved the first crystal structure for a group 2 SoxAX protein (SnSoxAX), where an N-terminal extension of SoxX forms a novel structure that supports dimer formation. Crystal structures of
SoxAX with a heme ligand substitution (C236M) uncovered an inherent flexibility of this SoxA heme site, with both bonding
distances and relative ligand orientation differing between asymmetric units and the new residue, Met236, representing an unusual rotamer of methionine. The flexibility of the SnSoxAXC236M SoxA heme environment is probably the cause of the four distinct, new EPR signals, including a high spin ferric heme form,
that were observed for the enzyme. Despite the removal of the catalytically active cysteine heme ligand and drastic changes
in the redox potential of the SoxA heme (WT, −479 mV; C236M, +85 mV), the substituted enzyme was catalytically active in glutathione-based
assays although with reduced turnover numbers (WT, 3.7 s−1; C236M, 2.0 s−1). SnSoxAXC236M was also active in assays using SoxYZ and thiosulfate as the sulfur substrate, suggesting that Cys236 aids catalysis but is not crucial for it. The SoxYZ-based SoxAX assay is the first assay for an isolated component of the
Sox multienzyme system.
Journal of Biological Chemistry 07/2011; 286(28):24872-24881. · 4.77 Impact Factor
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ABSTRACT: The synthesis and structural analysis (single crystal X-ray data) of two mononuclear ([Cu(L(1))(CN)]BF(4) and [Cu(L(3))(CN)](BF(4))) and three related, cyanide-bridged homodinuclear complexes ([{Cu(L(1))}(2)(CN)](BF(4))(3)·1.35 H(2)O, [{Cu(L(2))}(2)(CN)](BF(4))(3) and [{Ni(L(3))}(2)(CN)](BF(4))(3)) with a tetradentate (L(1)) and two isomeric pentadentate bispidine ligands (L(2), L(3); bispidines are 3,7-diazabicyclo[3.3.1]nonane derivatives) are reported, together with experimental magnetic, electron paramagnetic resonance (EPR), and electronic spectroscopic data and a ligand-field-theory-based analysis. The temperature dependence of the magnetic susceptibilities and EPR transitions of the dicopper(II) complexes, together with the simulation of the EPR spectra of the mono- and dinuclear complexes leads to an anisotropic set of g- and A-values, zero-field splitting (ZFS) and magnetic exchange parameters (Cu1: g(z) = 2.055, g(x) = 2.096, g(y) = 2.260, A(z) = 8, A(x) = 8, A(y) = 195 × 10(-4) cm(-1), Cu2: g and A as for Cu(1) but rotated by the Euler angles α = -6°, β = 100°, D(exc) = -0.07 cm(-1), E(exc)/D(exc) = 0.205 for [{Cu(L(1))}(2)(CN)](BF(4))(3)·1.35 H(2)O; Cu1,2: g(z) = 2.025, g(x) = 2.096, g(y) = 2.240, A(z) = 8, A(x) = 8, A(y) = 190 × 10(-4)cm(-1), D(exc) = -0.159 cm(-1), E(exc)/D(exc) = 0.080 for [{Cu(L(2))}(2)(CN)](BF(4))(3)). Thorough ligand-field-theory-based analyses, involving all micro states and all relevant interactions (Jahn-Teller and spin-orbit coupling) and DFT calculations of the magnetic exchange leads to good agreement between the experimental observations and theoretical predictions. The direction of the symmetric magnetic anisotropy tensor D(exc) in [{Cu(L(2))}(2)(CN)](BF(4))(3) is close to the Cu···Cu vector (22°), that is, nearly perpendicular to the Jahn-Teller axis of each of the two Cu(II) centers, and this reflects the crystallographically observed geometry. Antisymmetric exchange in [{Cu(L(1))}(2)(CN)](BF(4))(3)·1.35 H(2)O causes a mixing between the singlet ground state and the triplet excited state, and this also reflects the observed geometry with a rotation of the two Cu(II) sites around the Cu···Cu axis.
Inorganic Chemistry 06/2011; 50(15):6890-901. · 4.60 Impact Factor
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ABSTRACT: The synthesis and Cu(II) coordination chemistry of the cyclic pseudo-octapeptide H(4)pat(1), a dimethyl-imidazole analogue of naturally occurring cyclic peptides (patellamide A-F, ascidiacyclamide) is reported. Substitution of the oxazoline and thiazole heterocycles by dimethyl-imidazoles leads to a slightly different structure of the macrocycle in the solid state. The Cu(II) coordination chemistry of H(4)pat(1), monitored with high-resolution electrospray mass spectrometry, spectrophotometric titrations, and EPR spectroscopy, revealed the presence of both mono- and dinuclear Cu(II) complexes. The dimethyl-imidazole analogue shows a high cooperativity in Cu(II) coordination, that is, the preferred formation of dinuclear complexes. The dinuclear unbridged Cu(II) complexes of H(4)pat(1) have unusual EPR features, reminiscent of those of patellamide D: the dipole-dipole interaction of the Cu(II) centers is negligible due to the "magic angle" orientation of the two Cu(II) ions. Density functional theory calculations (DFT) are used to model the structures of the Cu(II) complexes, and the structural assignments from the spectroscopic investigations are supported by the optimized and by X-ray structures of the metal-free macrocycle and dinuclear Cu(II) complexes of H(4)pat(1). The rigidity of the dimethyl-imidazole rings has a significant effect on the structures of the metal-free ligands and Cu(II) complexes and therefore changes the properties of these compounds. This may explain why Nature has chosen the thiazole-oxazoline combination for the patellamides and ascidiacyclamide.
Inorganic Chemistry 06/2011; 50(11):5165-74. · 4.60 Impact Factor
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ABSTRACT: SoxAX cytochromes catalyze the formation of heterodisulfide bonds between inorganic sulfur compounds and a carrier protein, SoxYZ. They contain unusual His/Cys-ligated heme groups with complex spectroscopic signatures. The heme-ligating cysteine has been implicated in SoxAX catalysis, but neither the SoxAX spectroscopic properties nor its catalysis are fully understood at present. We have solved the first crystal structure for a group 2 SoxAX protein (SnSoxAX), where an N-terminal extension of SoxX forms a novel structure that supports dimer formation. Crystal structures of SoxAX with a heme ligand substitution (C236M) uncovered an inherent flexibility of this SoxA heme site, with both bonding distances and relative ligand orientation differing between asymmetric units and the new residue, Met(236), representing an unusual rotamer of methionine. The flexibility of the SnSoxAX(C236M) SoxA heme environment is probably the cause of the four distinct, new EPR signals, including a high spin ferric heme form, that were observed for the enzyme. Despite the removal of the catalytically active cysteine heme ligand and drastic changes in the redox potential of the SoxA heme (WT, -479 mV; C236M, +85 mV), the substituted enzyme was catalytically active in glutathione-based assays although with reduced turnover numbers (WT, 3.7 s(-1); C236M, 2.0 s(-1)). SnSoxAX(C236M) was also active in assays using SoxYZ and thiosulfate as the sulfur substrate, suggesting that Cys(236) aids catalysis but is not crucial for it. The SoxYZ-based SoxAX assay is the first assay for an isolated component of the Sox multienzyme system.
Journal of Biological Chemistry 05/2011; 286(28):24872-81. · 4.77 Impact Factor
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ABSTRACT: Sulfite dehydrogenase (SDH) from Starkeya novella is a heterodimeric enzyme comprising a Mo active site and a heme c electron relay, which mediates electron transfer from the Mo cofactor to cytochrome c following sulfite oxidation. Studies on the wild type enzyme (SDH(WT)) and its variants have identified key amino acids at the active site, specifically Arg-55 and His-57. We report the Mo(VI/V), Mo(V/IV) and Fe(III/II) (heme) redox potentials of the variants SDH(R55K), SDH(R55M), SDH(R55Q) and SDH(H57A) in comparison with those of SDH(WT). For SDH(R55M), SDH(R55Q) and SDH(H57A) the heme potentials are lowered from ca. 240mV in SDH(WT) to ca. 200mV, while the heme potential in SDH(R55K) remains unchanged and the Mo redox potentials are not affected significantly in any of these variants. Protein film voltammetry reveals a pH dependence of the electrochemical catalytic half-wave potential (E(cat)) of -59mV/pH in SDH(WT) and SDH(R55K) which tracks the pH dependence of the Mo(VI/V) redox potential. By contrast, the catalytic potentials for SDH(R55M) and SDH(H57A) are pH-independent and follow the potential of the heme cofactor. These results highlight a switch in the pathway of electron exchange as a function of applied potential that is revealed by protein film voltammetry where an actuation of rate limiting intramolecular electron transfer (IET, Mo to heme) at high potential attenuates the catalytic current relative to faster, direct electron transfer (Mo to electrode) at lower potential. The same change in electron transfer pathway is linked to an unusual peak-shaped profile of the ideally sigmoidal steady state voltammogram in SDH(WT) alone, which has been associated with a potential dependent change in the orientation of the enzyme on the electrode surface. All other variants show purely sigmoidal voltammetry due to their inherently slower turnover numbers which are always lower than IET rates.
Biochimica et Biophysica Acta 01/2011; 1807(1):108-18. · 4.66 Impact Factor
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Rosely A Peralta,
Adailton J Bortoluzzi,
Bernardo de Souza,
Rafael Jovito,
Fernando R Xavier,
Ricardo A. A. Couto,
Annelise Casellato,
Faruk Nome,
Andrew Dick,
Lawrence R. Gahan, [......],
Flavia C. S. de Paula,
Elene C Pereira-Maia,
Sergio de P. Machado,
Patricia C. Severino,
Claus Pich,
Tiago Bortolotto,
Hernan Terenzi,
Eduardo E Castellano,
Ademir Neves,
Mark J Riley
11/2010;
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Rosely A Peralta,
Adailton J Bortoluzzi,
Bernardo de Souza,
Rafael Jovito,
Fernando R Xavier,
Ricardo A A Couto,
Annelise Casellato,
Faruk Nome,
Andrew Dick,
Lawrence R Gahan, [......],
Flávia C S de Paula,
Elene C Pereira-Maia,
Sergio de P Machado,
Patricia C Severino,
Claus Pich,
Tiago Bortolotto,
Hernán Terenzi,
Eduardo E Castellano,
Ademir Neves,
Mark J Riley
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ABSTRACT: Purple acid phosphatases (PAPs) are a group of metallohydrolases that contain a dinuclear Fe(III)M(II) center (M(II) = Fe, Mn, Zn) in the active site and are able to catalyze the hydrolysis of a variety of phosphoric acid esters. The dinuclear complex [(H(2)O)Fe(III)(μ-OH)Zn(II)(L-H)](ClO(4))(2) (2) with the ligand 2-[N-bis(2-pyridylmethyl)aminomethyl]-4-methyl-6-[N'-(2-pyridylmethyl)(2-hydroxybenzyl) aminomethyl]phenol (H(2)L-H) has recently been prepared and is found to closely mimic the coordination environment of the Fe(III)Zn(II) active site found in red kidney bean PAP (Neves et al. J. Am. Chem. Soc. 2007, 129, 7486). The biomimetic shows significant catalytic activity in hydrolytic reactions. By using a variety of structural, spectroscopic, and computational techniques the electronic structure of the Fe(III) center of this biomimetic complex was determined. In the solid state the electronic ground state reflects the rhombically distorted Fe(III)N(2)O(4) octahedron with a dominant tetragonal compression aligned along the μ-OH-Fe-O(phenolate) direction. To probe the role of the Fe-O(phenolate) bond, the phenolate moiety was modified to contain electron-donating or -withdrawing groups (-CH(3), -H, -Br, -NO(2)) in the 5-position. The effects of the substituents on the electronic properties of the biomimetic complexes were studied with a range of experimental and computational techniques. This study establishes benchmarks against accurate crystallographic structural information using spectroscopic techniques that are not restricted to single crystals. Kinetic studies on the hydrolysis reaction revealed that the phosphodiesterase activity increases in the order -NO(2) ←Br ←H ←CH(3) when 2,4-bis(dinitrophenyl)phosphate (2,4-bdnpp) was used as substrate, and a linear free energy relationship is found when log(k(cat)/k(0)) is plotted against the Hammett parameter σ. However, nuclease activity measurements in the cleavage of double stranded DNA showed that the complexes containing the electron-withdrawing -NO(2) and electron-donating -CH(3) groups are the most active while the cytotoxic activity of the biomimetics on leukemia and lung tumoral cells is highest for complexes with electron-donating groups.
Inorganic Chemistry 11/2010; 49(24):11421-38. · 4.60 Impact Factor
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ABSTRACT: The CuII coordination chemistry of three synthetic analogues of westiellamide (H3Lwa) with an [18]azacrown-6 macrocyclic structure and imidazole (H3L1), oxazole (H3L2), or thiazole (H3L3) heterocyclic donors in addition to the peptide groups, is reported. The Nheterocycle-Npeptide-N(heterocycle) binding sites are highly preorganized for the coordination to CuII ions. The stability constants of mono- and dinuclear CuII complexes of H3L1, H3L2, and H3L3, obtained by isothermal titration microcalorimetry, are reported. EPR and NMR spectroscopy as well as electrospray ionization mass spectrometry (ESI-MS) were used to characterize the complexes formed in solution. The stabilities of the mononuclear and dinuclear CuII complexes of the three ligands are in the range of 10(5) M(-1), but there are subtle differences; specifically the oxazole-derived ligand has, in contrast to the other two macrocycles, a negative formation entropy for coordination to the first CuII ion and a higher stability for complexation to a second CuII center in comparison with the first CuII center (cooperativity). Differences between the three ligands are also apparent in terms of the formation mechanism. With the oxazole-based ligand H3L2, NMR spectroscopy, EPR spectroscopy, and ESI-MS indicate the formation of a ligand-CuII 2:1 intermediate, and this may explain the differences in the formation entropy as well as the cooperativity.
European Journal of Biochemistry 09/2010; 15(7):1129-35. · 3.42 Impact Factor
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Zhen Li,
Lina Cheng,
Qiao Sun,
Zhonghua Zhu,
Mark J Riley,
Muhsen Aljada,
Zhenxiang Cheng,
Xiaolin Wang, Graeme R Hanson,
Shizhang Qiao,
Sean C Smith,
Gao Qing Max Lu
Angewandte Chemie International Edition 03/2010; 49(15):2777-81. · 13.45 Impact Factor
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ABSTRACT: While manganese has been successfully exploited as a spectroscopic probe of EPR silent centers (Zn, Ca, Mg) in metalloenzymes,
it was only during the last decade that manganese-containing metalloenzymes were investigated in great detail. Indeed, in
some biological systems it remains unclear whether iron and/or manganese is required for catalytic competency. Binuclear manganese
enzymes are a small group of enzymes that catalyze a variety of chemical reactions and are involved in numerous metabolic
functions. In this review the structural and biochemical properties of these enzymes are described. The contributions of electron
paramagnetic resonance-related techniques to our understanding of the structure and reactivity of binuclear manganese enzymes
are discussed and, where appropriate, supported by data obtained from complementary spectroscopic methods. This article is
intended as a guide to illustrate the usefulness of electron paramagnetic resonancerelated techniques in the study of these
enzymes.
12/2009: pages 273-341;
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ABSTRACT: The binuclear heterovalent manganese model complex [Mn(II)Mn(III)(L1)(OAc)(2)] ClO(4) x H(2)O (H(2)L1 = 2-(((3-((bis(pyridin-2-ylmethyl)amino)methyl)-2-hydroxy-5-methylbenzyl)(pyridin-2-ylmethyl)amino)-methyl)phenol) has been prepared and studied structurally, spectroscopically, and computationally. The magnetic and electronic properties of the complex have been related to its structure. The complex is weakly antiferromagnetically coupled (J approximately -5 cm(-1), H = -2J S(1) x S(2)) and the electron paramagnetic resonance (EPR) and magnetic circular dichroism (MCD) spectra identify the Jahn-Teller distortion of the Mn(III) center as predominantly a tetragonal compression, with a significant rhombic component. Electronic structure calculations using density functional theory have confirmed the conclusions derived from the experimental investigations. In contrast to isostructural M(II)Fe(III) complexes (M = Fe, Mn, Zn, Ni), the Mn(II)Mn(III) system is bifunctional possessing both catalase and hydrolase activities, and only one catalytically relevant pK(a) (= 8.2) is detected. Mechanistic implications are discussed.
Inorganic Chemistry 11/2009; 48(21):10036-48. · 4.60 Impact Factor
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Pia Nielsen,
Hans Toftlund,
Andrew D Bond,
John F Boas,
John R Pilbrow, Graeme R Hanson,
Christopher Noble,
Mark J Riley,
Suzanne M Neville,
Boujemaa Moubaraki,
Keith S Murray
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ABSTRACT: A family of spin crossover cobalt(II) complexes of the type [Co(terpyRX)(2)](Y)(2) x nH(2)O (X = 4, 8, 12 and Y = BF(4)(-), ClO(4)(-), PF(6)(-), BPh(4)(-)) has been synthesized, whereby the alkyl chain length, RX, and counteranion, Y, have been systematically varied. The structural (single crystal X-ray diffraction) and electronic (magnetic susceptibility, electron paramagnetic resonance (EPR)) properties have been investigated within this family of compounds. Single crystal X-ray diffraction analysis of [Co(terpyR8)(2)](ClO(4))(2), [Co(terpyR8)(2)](BF(4))(2) x H(2)O, and [Co(terpyR4)(2)](PF(6))(2) x 3 H(2)O, at 123 K, revealed compressed octahedral low spin Co(II) environments and showed varying extents of disorder in the alkyl tail portions of the terpyRX ligands. The magnetic and EPR studies were focused on the BF(4)(-) family and, for polycrystalline solid samples, revealed that the spin transition onset temperature (from low to high spin) decreased as the alkyl chain lengthened. EPR studies of polycrystalline powder samples confirmed these results, showing signals only due to the low spin state at the temperatures seen in magnetic measurements. Further to this, simultaneous simulation of the EPR spectra of frozen solutions of [Co(terpyR8)(2)](BF(4))(2) x H(2)O, recorded at S-, X-, and Q-band frequencies, allowed accurate determination of the g and A values of the low spin ground state. The temperature dependence of the polycrystalline powder EPR spectra of this and the R4 and R12 complexes is explained in terms of Jahn-Teller effects using the warped Mexican hat potential energy surface model perturbed by the low symmetry of the ligands. While well recognized in Cu(II) systems, this is one of the few times this approach has been used for Co(II).
Inorganic Chemistry 09/2009; 48(15):7033-47. · 4.60 Impact Factor
