Biomedical subjects
Publications and source records attributed to Martin L Kirk.
The {DyIII[15-MC-CuII(N)(S)-pheHA(-5)]}3+ complex displays slow magnetic relaxation behavior in a frozen solution at low temperature, whereas the analogous HoIII structure does not exhibit similar behavior.
S-Ribosylhomocysteinase (LuxS) catalyzes the cleavage of the thioether linkage in S-ribosylhomocysteine (SRH) to produce homocysteine (Hcys) and 4,5-dihydroxy-2,3-pentanedione (DPD), the precursor of type II bacterial autoinducer (AI-2). The proposed catalytic mechanism involves two consecutive ribose carbonyl migration steps via an intramolecular redox reaction and a subsequent beta-elimination step, all catalyzed by a divalent metal ion (e.g., Fe(2+) or Co(2+)) and two general acids/bases in the active site. Absorption and EPR spectroscopic studies were performed with both wild-type and various mutant forms of LuxS under a wide range of pH conditions. The studies revealed a pK(a) of 10.4 for the metal-bound water. The pK(a) value of Cys-83 was determined to be <6 by (13)C-(1)H HSQC NMR experiments with [3-(13)C]cysteine-labeled Zn(2+)-substituted Escherichia coli LuxS. The active form of LuxS contains a metal-bound water and a thiolate ion at Cys-83, consistent with the proposed roles of the metal ion (Lewis acid) and Cys-83 (general acid/base) during catalysis. Finally, an invariant Arg-39 in the active site was demonstrated to be at least partially responsible for stabilizing the thiolate anion of Cys-83.
Two structurally characterized dinuclear valence tautomers are described. Cobalt ions are bridged by p- and m-phenylene units connected to 2,2'-bipyridines. X-ray crystal structures show that the molecules are in the [(Co(III))(Co(III))] forms at ca. 125 K, while spectroscopic studies show that both molecules can achieve the [(Co(II))(Co(II))] form above 400 K and confirm the [(Co(III))(Co(III))] form below 10 K. Magnetic susceptibility studies are also included. Our results highlight the necessity of studying both crystalline and amorphous samples to distinguish the effects of intrinsic electronic structure and intermolecular forces on valence tautomeric behavior.
New paramagnetic, heterobimetallic Mo/Cu complexes featuring the Mo(=O)(mu-S)Cu core of O. carboxidovorans carbon monoxide dehydrogenase have been synthesized and structurally and spectroscopically characterized. The complexes exhibit EPR spectra (left graphic) indicative of extensive electron delocalization across the Mo-S-Cu core, in agreement with computational studies identifying the singly-occupied molecular orbital (right graphic).
X-ray crystallography and resonance Raman (rR) spectroscopy have been used to further characterize (Tp*)MoO(qdt) (Tp* is hydrotris(3,5-dimethyl-1-pyrazolyl)borate and qdt is 2,3-quinoxalinedithiolene), which represents an important benchmark oxomolybdenum mono-dithiolene model system relevant to various pyranopterin Mo enzyme active sites, including sulfite oxidase. The compound (Tp*)MoO(qdt) crystallizes in the triclinic space group, P1, where a = 9.8424 (7) A, b = 11.2323 (8) A, c = 11.9408 (8) A, alpha = 92.7560 (10) degrees, beta = 98.9530 (10) degrees, and gamma = 104.1680 (10) degrees. The (Tp*)MoO(qdt) molecule exhibits the distorted six-coordinate geometry characteristic of related oxo-Mo(V) systems possessing a single coordinated dithiolene ligand. The first coordination sphere bond lengths and angles in (Tp*)MoO(qdt) are very similar to the corresponding structural parameters for (Tp*)MoO(bdt) (bdt is 1,2-benzenedithiolene). The relatively small inner-sphere structural variations observed between (Tp*)MoO(qdt) and (Tp*)MoO(bdt) strongly suggest that geometric effects are not a major contributor to the significant electronic structural differences reported for these two oxo-Mo(V) dithiolenes. Therefore, the large differences observed in the reduction potential and first ionization energy between the two molecules appear to derive primarily from differences in the effective nuclear charges of their respective sulfur donors. However, a subtle perturbation to Mo-S bonding is implied by the nonplanarity of the dithiolene chelate ring, which is defined by the fold angle. This angular distortion (theta = 29.5 degrees in (Tp*)MoO(qdt); 21.3 degrees in (Tp*)MoO(bdt)) observed between the MoS2 and S-C=C-S planes may contribute to the electronic structure of these oxo-Mo dithiolene systems by controlling the extent of S p-Mo d orbital overlap. In enzymes, the fold angle may be dynamically modulated by the pyranopterin, thereby functioning as a transducer of vibrational energy associated with protein conformational changes directly to the active site via changes in the fold angle. This process could effectively mediate charge redistribution at the active site during the course of atom- and electron-transfer processes. The rR spectrum shows bands at 348 and 407 cm(-1). From frequency analysis of the normal modes of the model, [(NH3)3MoO(qdt)]1+, using the Gaussian03 suite of programs, these bands are assigned as mixed-mode Mo-S vibrations of the five-membered Mo-ditholene core structure. Raman spectroscopy has also provided additional evidence for an in-plane pseudo-sigma dithiolene S-Mo d(xy) covalent bonding interaction in (Tp*)MoO(qdt) and related oxo-Mo-dithiolenes that has implications for electron-transfer regeneration of the active site in sulfite oxidase involving the pyranopterin dithiolene.
Plant sulfite oxidase from Arabidopsis thaliana has been characterized both spectroscopically and kinetically. The enzyme is unusual in lacking the heme domain that is present in the otherwise highly homologous enzyme from vertebrate sources. In steady-state assays, the enzyme exhibits a pH maximum of 8.5 and is also found to function as a selenite oxidase. Sulfite at the lowest experimentally feasible concentrations reduces the enzyme within the dead-time of a stopped-flow instrument at 5 degrees C, indicating that the A. thaliana enzyme has a limiting rate constant for reduction, k(red), at least 10 times greater than that of the chicken enzyme (190 s(-1)). The EPR parameters for the high- and low-pH forms of the A. thaliana enzyme have been determined, and the g-values are found to resemble those previously reported for the vertebrate enzymes. Finally, the A. thaliana enzyme has been probed by resonance Raman spectroscopy. A detailed analysis of the vibrational spectrum in the region where Mo=O stretching modes are anticipated to occur has been performed with the help of density functional theory calculations, evaluated in the context of the Raman data. Calculated frequencies obtained for two model systems have been compared to experimental resonance Raman spectra of oxidized A. thaliana sulfite oxidase catalytically cycled in both H2(16)O and H2(18)O. The vibrational frequency shifts observed upon (18)O-labeling of the enzyme are consistent with theoretical models in which either the equatorial oxygen or both equatorial and axial atoms of the dioxomolybdenum center are labeled. Importantly, the vibrational mode description is consistent with the active site possessing geometrically inequivalent oxo ligands and a Mo d(xy) redox-active molecular orbital oriented in the equatorial plane forming a pi-bonding interaction solely with the equatorial oxo, O(eq). Electron occupancy of this Mo=O(eq) pi* redox orbital upon interaction with substrates would effectively labilize the Mo=O(eq) bond, providing the dominant contribution to lowering the activation energy for oxygen atom transfer.
The effect of sterically encumbering ligands on the electronic structure of oxomolybdenum tetrathiolate complexes was determined using a combination of electronic absorption and magnetic circular dichroism spectroscopies, complimented by DFT bonding calculations, to understand geometric and electronic structure contributions to reduction potentials. These complexes are rudimentary models for a redox-active metalloenzyme active site in a protein matrix and allow for detailed spectroscopic probing of specific oxomolybdenum-thiolate interactions that are directly relevant to Mo-S(cysteine) bonding in pyranopterin molybdenum enzymes. Data are presented for three para-substituted oxomolybdenum tetrathiolate complexes ([PPh4][MoO(p-SPhCONHCH3)4], [PPh4][MoO(p-SPhCONHC(CH2O(CH2)2CN)3)4], and [PPh4][MoO(p-SPhCONHC(CH2O(CH2)2COOCH2CH3)3)4]). The Mo(V/IV) reduction potentials of the complexes in DMF are -1213, -1251, and -1247 mV, respectively. The remarkably similar electronic absorption and magnetic circular dichroism spectra of these complexes establish that the observed reduction potential differences are not a result of significant changes in the electronic structure of the [MoOS4]- cores as a function of the larger ligand size. We provide evidence that these reduction potential differences result from the driving force for a substantial reorganization of the O-Mo-S-C dihedral angle upon reduction, which decreases electron donation from the thiolate sulfurs to the reduced molybdenum center. The energy barrier to favorable O-Mo-S-C geometries results in a reorganizational energy increase, relative to [MoO(SPh)4](-/2-), that correlates with ligand size. The inherent flexible nature of oxomolybdenum-thiolate bonds indicate that thiolate ligand geometry, which controls Mo-S covalency, could affect the redox processes of monooxomolybdenum centers in pyranopterin molybdenum enzymes.
The structural characterization of complexes [Mn(II)4Mn(III)22(pdol)12(OCH3)12(O)16(N3)6] (1) and [Mn(II)4Mn(III)22(pdol)12(OCH3)12(O)16(OH)2(H3O)(OCH3)3].ClO4.5CH3OH (2), where pdol(2-) is di-2-pyridyl methanediol, reveals that each has a metallacryptand shell that encapsulates a manganese oxide core. Variable-temperature direct current magnetic susceptibility measurements on 2 indicate a paramagnetic ground state that results from an overall antiferromagnetic interaction in the cluster, with chiT values decreasing from 300 K (51.2 cm3 K mol(-1)) to 2 K (19.8 cm3 K mol(-1)). Variable-temperature alternating current magnetic susceptibility measurements imply that both 1 and 2 behave as single-molecule magnets. Fitting the frequency-dependent out-of-phase magnetic susceptibility to the Arrhenius equation yields an effective energy barrier, Ueff, to magnetization relaxation of 16.5 +/- 0.7 K (11.5 +/- 0.5 cm(-1)) for 1 and 36.2 +/- 2.0 K (25.1 +/- 1.4 cm(-1)) for 2. The larger value for 2 is in agreement with the lower molecular symmetry, larger magnetoanisotropy, and higher ground spin state of 2 compared to those of 1. This observation suggests a new strategy for increasing the blocking temperatures in high-nuclearity manganese clusters.
A terminal Pd-oxo unit is reported. The unit is encapsulated in a cavity defined by two [A-alpha-PW9O34]9- units fused together by one [WO(OH2)]4+ center and forms from Pd(II) in buffered media in the presence of O2. Both X-ray diffraction and EXAFS data are consistent with a Pd-oxo bond distance of ca. 1.65 A. 17O NMR studies confirm that the solid-state structure is maintained in solution.
Terminal mono-oxo complexes of the late transition metal elements have long been considered too unstable to synthesize because of repulsion between the oxygen electrons and the mostly filled metal d orbitals. A platinum(IV)-oxo compound flanked by two polytungstate ligands, K7Na9[O=Pt(H2O)L2], L = [PW9O34(9-)], has now been prepared and isolated at room temperature as air-stable brown crystals. X-ray and neutron diffraction at 30 kelvin revealed a very short [1.720(18) angstrom] Pt-O bond and no evidence of a hydrogen atom at the terminal oxygen, ruling out a better precedented Pt-OH complex. Density functional theory and spectroscopic data account for the stability of the Pt(IV)-oxo unit by electron withdrawal into delocalized orbitals of the polytungstates.
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The electronic structure of cis,trans-(L-N(2)S(2))MoO(X) (where L-N(2)S(2) = N,N'-dimethyl-N,N'-bis(2-mercaptophenyl)ethylenediamine and X = Cl, SCH(2)C(6)H(5), SC(6)H(4)-OCH(3), or SC(6)H(4)CF(3)) has been probed by electronic absorption, magnetic circular dichroism, and resonance Raman spectroscopies to determine the nature of oxomolybdenum-thiolate bonding in complexes possessing three equatorial sulfur ligands. One of the phenyl mercaptide sulfur donors of the tetradentate L-N(2)S(2) chelating ligand, denoted S(180), coordinates to molybdenum in the equatorial plane such that the OMo-S(180)-C(phenyl) dihedral angle is approximately 180 degrees, resulting in a highly covalent pi-bonding interaction between an S(180) p orbital and the molybdenum d(xy) orbital. This highly covalent bonding scheme is the origin of an intense low-energy S --> Mo d(xy) bonding-to-antibonding LMCT transition (E(max) approximately 16000 cm(-)(1), epsilon approximately 4000 M(-)(1) cm(-)(1)). Spectroscopically calibrated bonding calculations performed at the DFT level of theory reveal that S(180) contributes approximately 22% to the HOMO, which is predominantly a pi antibonding molecular orbital between Mo d(xy) and the S(180) p orbital oriented in the same plane. The second sulfur donor of the L-N(2)S(2) ligand is essentially nonbonding with Mo d(xy) due to an OMo-S-C(phenyl) dihedral angle of approximately 90 degrees. Because the formal Mo d(xy) orbital is the electroactive or redox orbital, these Mo d(xy)-S 3p interactions are important with respect to defining key covalency contributions to the reduction potential in monooxomolybdenum thiolates, including the one- and two-electron reduced forms of sulfite oxidase. Interestingly, the highly covalent Mo-S(180) pi bonding interaction observed in these complexes is analogous to the well-known Cu-S(Cys) pi bond in type 1 blue copper proteins, which display electronic absorption and resonance Raman spectra that are remarkably similar to these monooxomolybdenum thiolate complexes. Finally, the presence of a covalent Mo-S pi interaction oriented orthogonal to the MOO bond is discussed with respect to electron-transfer regeneration in sulfite oxidase and Mo=S(sulfido) bonding in xanthine oxidase.
A new class of oxo-bridged high-valent hexamanganese (Mn6) clusters containing a novel (Mn6O8)6+ core, [MnIV(4)MnIII2(mu-O)4(mu3-O)4(dmb)6(O2CR)2]4+ (where dmb=4,4'-dimethyl-2,2'-bipyridine, and RCO2=2,6-di(p-tolyl)benzoate (Ar(Tol)CO2-) (3) or 2,6-di(4-tert-butylphenyl)benzoate (Ar(4-tBuPh)CO2-) (4)), was synthesized using sterically hindered m-terphenyl-derived carboxylate ligands. These complexes can be synthesized by oxidizing the MnII mononuclear complexes, [Mn(dmb)2(OH2)(O2CR)]+ (where RCO2=Ar(Tol)CO2- (1) or Ar(4-tBuPh)CO2- (2)) with (n-Bu4N)MnO4, by direct Mn(II) + Mn(VII) in situ comproportionation reactions, or by ligand substitution on the dinuclear manganese (III,IV) or (IV,IV) complexes, [(Mn2(mu-O)2(dmb)4)](3+/4+). The compound [MnIV4MnIII2(mu-O)4(mu3-O)4(dmb)6(Ar(Tol)CO2)2](OTf)4 [3(OTf)4] crystallizes in the monoclinic space group P2(1)/n, with the cell parameters a=15.447(1) A, b=15.077(2) A, c=27.703(2) A, beta=91.68(2) degrees, V=6449.3(6) A3, and Z=2. The X-ray structure reveals that there are three different bridging modes for the oxo groups: mu, "pyramidal" mu3, and "T-shaped" mu3. Solid-state variable temperature magnetic susceptibility studies suggest that the Mn centers are net antiferromagnetically coupled to yield a diamagnetic ST=0 ground spin state with a large number of low-lying, thermally accessible states with ST>0. 1H NMR spectra were recorded for both Mn6 clusters and selected resonances assigned. The electronic and redox properties of these complexes along with the effect of the presence of the bulky carboxylate ligands are also described here.
Deconvoluting the different contributions of thiolate and ene-1,2-dithiolate donors to the underlying electronic structure of the Mo site in sulfite oxidase (SO) has proven to be a difficult task. One way in which these differences might be illuminated is by selectively substituting Se for S in model complexes which possess multiple sulfur donor ligand environments. Here we report the synthesis and structures of new oxo-Mo(V) complexes as effective models for the one-electron reduced active site of SO. We have used the tridentate heteroscorpionate ligand (2-dimethylethanethiol)bis(3,5-dimethylpyrazolyl)methane (L3SH) in order to model the constrained cysteinyl sulfur (S(Cys)) ligand environment observed in the crystal structure of the enzyme, and benzene-1,2-dithiol (bdt) as a mimic of the ene-1,2-dithiolate chelate. [(L3S)MoO(bdt)] and [(L3S)MoO(SPh)(2)] have been structurally characterized by X-ray crystallography, and as such, [(L3S)MoO(bdt)] is only the second known model compound that closely approximates the active site structure of reduced forms of SO. Additionally, benzenethiol (SPh) and benzeneselenol (SePh) have been used to perturb the equatorial ligand environment of [(L3S)MoO(bdt)].) This has provided much needed insight into the electronic structure of the one-electron reduced SO site and has allowed for increased understanding of the individual roles played by these different thiolate donors in the oxidative half-reaction of the enzyme. Interestingly, the EPR spectra of [(L3S)MoO(bdt)], [(L3S)MoO(SPh)(2)], and [(L3S)MoO(SePh)(2)] closely resemble that of both high pH (hpH) and low pH (lpH) SO, except for the fact that the magnitude of g(1) is found to be consistently higher in the model spectra compared to that of the enzyme. It is suggested that this derives from an increase in Mo-S covalency in the models relative to hpH and lpH SO.
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Binuclear manganese complexes Mn2(III/IV)(dtsalpn)2DCBI, 1, Mn2(III/III)(dtsalpn)2HDCBI, 2, containing the ligand dicarboxyimidazole (DCBI) have been prepared in order to address the issue of imidazole bridged and ferromagnetically coupled Mn sites in high oxidation states of the OEC in Photosystem II (PS II). Temperature dependent magnetic susceptibility studies of 1 indicates that the interaction between the two Mn(III)/Mn(IV) ions is ferromagnetic (J = +1.4 cm(-1)). Variable temperature EPR spectra of 1 shows that a g = 2 multiline is as an excited state signal corresponding to S = 1/2.
We report molecular structures and temperature-dependent magnetic susceptibility data for several new metal complexes of heterospin triplet ground-state biradical ligands. The ligands are comprised of both nitronyl-nitroxide (NN) and semiquinone (SQ) spin carriers. Five compounds are five-coordinate M(II) complexes (M = Mn, Co, Ni, Cu, and Zn), and one is a six-coordinate Ni(II) complex. Five compounds were structurally characterized. During copper complex formation a reaction with methanol occurs to form a unique methoxy-substituted SQ ring. Variable-temperature magnetic susceptibility studies are consistent with strong intraligand (NN-SQ and NN-PhSQ) ferromagnetic exchange coupling. For the five-coordinate Mn, Co, and Ni complexes, the S = 1 ligand is antiferromagnetically coupled to the metal. For both the five-coordinate Cu complex and the six-coordinate Ni complex, the ligand is ferromagnetically coupled to the metal spins in accordance with orbital symmetry arguments. Despite the low molecular symmetries, the predicted trend in metal-ligand exchange interactions is supported by spin dimer analysis based on extended Hückel calculations. For (NN-SQ)NiTp(Cum,Me)() (Tp(Cum,Me)() = hydro-tris(3-cumenyl-5-methylpyrazolyl)borate), an antisymmetric exchange term was required for the best fit of the magnetic susceptibility data. Antisymmetric exchange was less important for the other complexes due to inherently smaller Deltag. Finally, it is shown that intraligand exchange coupling is of paramount importance in stabilizing high-spin states of mixed metal-biradical complexes.
Magnetic circular dichroism (MCD) continues to be a powerful probe of metalloenzyme electronic and geometric structure, in addition to playing a major role in the determination of heme enzyme coordination geometries. Excited state electronic structure contributions to enzyme activity have been gleaned from C-term MCD studies, which are usually interpreted in the context of other spectroscopies, including electronic absorption and resonance Raman. The recent development of sophisticated methods for the analysis of variable-temperature, variable-field MCD have allowed the ground states of metalloenzyme active sites to be studied in detail, providing information on the electronic and geometric structure of the site. This has been especially informative for non-heme iron enzymes. In the past two years X-ray MCD has been shown to be a promising technique for the study of metalloenzymes.