The Mn(2+)-bicarbonate complex in a frozen solution revisited by pulse W-band ENDOR

The coordination of bicarbonate to Mn (2+) is the simplest model system for the coordination of Mn (2+) to carboxylate residues in a protein. Recently, the structure of such a complex has been investigated by means of X-band pulse EPR (electron paramagnetic resonance) experiments ( Dasgupta, J. ; et al. J. Phys. Chem. B 2006, 110, 5099 ). Based on the EPR results, together with electrochemical titrations, it has been concluded that the Mn (2+) bicarbonate complex consists of two bicarbonate ligands, one of which is monodentate and other bidentate, but only the latter has been observed by the pulsed EPR techniques. The X-band measurements, however, suffer several drawbacks. (i) The zero-field splitting (ZFS) term of the spin Hamiltonian affects the nuclear frequencies. (ii) There are significant contributions from ENDOR (electron nuclear double resonance) lines of the M S not equal +/- (1)/ 2 manifolds. (iii) There are overlapping signals of (23)Na. All these reduce the uniqueness of the data interpretation. Here we present a high-field ENDOR investigation of Mn (2+)/NaH (13)CO 3 in a water/methanol solution that eliminates the above difficulties. Both Davies and Mims ENDOR measurements were carried out. The spectra show that a couple of slightly inequivalent (13)C nuclei are present, with isotropic and anisotropic hyperfine couplings of A iso1 = 1.2 MHz, T perpendicular1 = 0.7 MHz, A iso2 = 1.0 MHz, T perpendicular2 = 0.6 MHz, respectively. The sign of the hyperfine coupling was determined by variable mixing time (VMT) ENDOR measurements. These rather close hyperfine parameters suggest that there are either two distinct, slightly different, carbonate ligands or that there is some distribution in conformation in only one ligand. The distances extracted from T perpendicular1 and T perpendicular2 are consistent with a monodentate binding mode. The monodentate binding mode and the presence of two ligands were further supported by DFT calculations and (1)H ENDOR measurements. Additionally, (23)Na ENDOR resolved at least two types of (23)Na (+) in the Mn (2+)-bicarbonate complex, thus suggesting that the bicarbonate bridges two positively charged metal ions.     

55Mn pulse ENDOR at 34 GHz of the S0 and S2 states of the oxygen-evolving complex in photosystem II

55Mn pulse ENDOR experiments at 34 GHz (Q-band) are reported for the S0 and S2 states of the oxygen-evolving complex of photosystem II. Their numerical analysis (i) shows that in both states all four Mn ions are magnetically coupled, (ii) allows a refinement of the hyperfine interaction (HFI) parameters obtained earlier for the S2 state at X-band (Peloquin, J. M.; Campbell, K. A.; Randall, D. W.; Evanchik, M. A.; Pecoraro, V. L.; Armstrong, W. H.; Britt, R. D. J. Am. Chem. Soc. 2000, 122, 10926-10942), (iii) provides the first reliable 55Mn HFI tensors for the S0 state, and (iv) leads to the suggestion that the Mn oxidation states in S0 and S2 are Mn4(III, III, III, IV) and Mn4(III, IV, IV, IV), respectively. In addition, a Q-band EPR spectrum is reported for the S0 state, and inversion-recovery experiments at 4.5 K directly show that the electron spin-lattice relaxation for the S0 state is about 2 orders of magnitude faster than that for the S2 state.     

The catalytic Mn2+ sites in the enolase-inhibitor complex: crystallography, single-crystal EPR, and DFT calculations

Crystals of Zn2+/Mn2+ yeast enolase with the inhibitor PhAH (phosphonoacetohydroxamate) were grown under conditions with a slight preference for binding of Zn2+ at the higher affinity site, site I. The structure of the Zn2+/Mn2+-PhAH complex was solved at a resolution of 1.54 A, and the two catalytic metal binding sites, I and II, show only subtle displacement compared to that of the corresponding complex with the native Mg2+ ions. Low-temperature echo-detected high-field (W-band, 95 GHz) EPR (electron paramagnetic resonance) and 1H ENDOR (electron-nuclear double resonance) were carried out on a single crystal, and rotation patterns were acquired in two perpendicular planes. Analysis of the rotation patterns resolved a total of six Mn2+ sites, four symmetry-related sites of one type and two out of the four of the other type. The observation of two chemically inequivalent Mn2+ sites shows that Mn2+ ions populate both sites I and II and the zero-field splitting (ZFS) tensors of the Mn2+ in the two sites were determined. The Mn2+ site with the larger D value was assigned to site I based on the 1H ENDOR spectra, which identified the relevant water ligands. This assignment is consistent with the seemingly larger deviation of site I from octahedral symmetry, compared to that of site II. The ENDOR results gave the coordinates of the protons of two water ligands, and adding them to the crystal structure revealed their involvement in a network of H bonds stabilizing the binding of the metal ions and PhAH. Although specific hyperfine interactions with the inhibitor were not determined, the spectroscopic properties of the Mn2+ in the two sites were consistent with the crystal structure. Density functional theory (DFT) calculations carried out on a cluster representing the catalytic site, with Mn2+ in site I and Zn2+ in site II, and vice versa, gave overestimated D values on the order of the experimental ones, although the larger D value was found for Mn2+ in site II rather than in site I. This discrepancy was attributed to the high sensitivity of the ZFS parameters to the Mn-O bond lengths and orientations, such that small, but significant, differences between the optimized and crystal structures alter the ZFS considerably, well above the difference between the two sites.     

Ultrahigh-field NMR spectroscopy of quadrupolar transition metals: 55Mn NMR of several solid manganese carbonyls

55Mn NMR spectra acquired at 21.14 T (nu(L)(55Mn) = 223.1 MHz) are presented and demonstrate the advantages of using ultrahigh magnetic fields for characterizing the chemical shift tensors of several manganese carbonyls: eta5-CpMn(CO)3, Mn2(CO)10, and (CO)5MnMPh3 (M = Ge, Sn, Pb). For the compounds investigated, the anisotropies of the manganese chemical shift tensors are less than 250 ppm except for eta5-CpMn(CO)3, which has an anisotropy of 920 ppm. At 21.14 T, one can excite the entire m(I) = 1/2 <--> m(I) = -1/2 central transition of eta5-CpMn(CO)3, which has a breadth of approximately 700 kHz. The breadth arises from second-order quadrupolar broadening due to the 55Mn quadrupolar coupling constant of 64.3 MHz, as well as the anisotropic shielding. Subtle variations in the electric field gradient tensors at the manganese are observed for crystallographically unique sites in two of the solid pentacarbonyls, resulting in measurably different C(Q) values. MQMAS experiments are able to distinguish four magnetically unique Mn sites in (CO)(5)MnPbPh3, each with slightly different values of delta(iso), C(Q), and eta(Q).     

Single-crystal 55Mn NMR spectra of two Mn12 single-molecule magnets

The initial application is reported of single-crystal 55Mn NMR spectroscopy, and associated orientation dependence studies, to single-molecule magnets (SMMs). The studies were performed on two members of the Mn12 family of SMMs, [Mn12O12(O2CMe)16(H2O)4].2MeCO2H.4H2O (Mn12-Ac) and [Mn12O12(O2CCH2Br)16(H2O)4].4CH)Cl) (Mn12-BrAc). Single-crystal spectra give a dramatic improvement in the spectral resolution over oriented powder spectra, allowing the clear observation of quadrupolar splittings, the determination of quadrupole coupling parameters (e2qQ), and an assessment of the symmetry-lowering perturbation of the core of Mn12-Ac by hydrogen-bonding interactions with lattice solvate molecules of crystallization. The results emphasize the utility of single-crystal NMR studies to probe the cores of these magnetic molecules.     

Computation of hyperfine tensors for dinuclear Mn(III) Mn(IV) complexes by broken-symmetry approaches: anisotropy transfer induced by local zero-field splitting

Based on broken-symmetry density functional calculations, the (55)Mn hyperfine tensors of a series of exchange-coupled, mixed-valence, dinuclear Mn(III) Mn(IV) complexes have been computed. We go beyond previous quantum chemical work by fully including the effects of local zero-field splitting (ZFS) interactions in the spin projection, following the first-order perturbation formalism of Sage et al. [J. Am. Chem. Soc. 1989, 111, 7239]. This allows the ZFS-induced transfer of hyperfine anisotropy from the Mn(III) site to the Mn(IV) site to be described with full consideration of the orientations of local hyperfine and ZFS tensors. After scaling to correct for systematic deficiencies in the quantum chemically computed local ZFS tensors, good agreement with experimental (55)Mn anisotropies at the Mn(IV) site is obtained. The hyperfine coupling anisotropies on the Mn(III) site depend sensitively on structural distortions for a d(4) ion. The latter are neither fully reproduced by using a DFT-optimized coordination environment nor by using experimental structures. For very small exchange-coupling constants, the perturbation treatment breaks down and a dramatic sensitivity to the scaling of the local ZFS tensors is observed. These results are discussed with respect to ongoing work to elucidate the structure of the oxygen-evolving complex of photosystem II by analysis of the EPR spectra.     

A solid-state 55Mn NMR spectroscopy and DFT investigation of manganese pentacarbonyl compounds

Central transition (55)Mn NMR spectra of several solid manganese pentacarbonyls acquired at magnetic field strengths of 11.75, 17.63, and 21.1 T are presented. The variety of distinct powder sample lineshapes obtained demonstrates the sensitivity of solid-state (55)Mn NMR to the local bonding environment, including the presence of crystallographically unique Mn sites, and facilitates the extraction of the Mn chemical shift anisotropies, CSAs, and the nuclear quadrupolar parameters. The compounds investigated include molecules with approximate C(4v) symmetry, LMn(CO)(5)(L = Cl, Br, I, HgMn(CO)(5), CH(3)) and several molecules of lower symmetry (L = PhCH(2), Ph(3-n)Cl(n)Sn (n= 1, 2, 3)). For these compounds, the Mn CSA values range from <100 ppm for Cl(3)SnMn(CO)(5) to 1260 ppm for ClMn(CO)(5). At 21.1 T the (55)Mn NMR lineshapes are appreciably influenced by the Mn CSA despite the presence of significant (55)Mn quadrupolar coupling constants that range from 8.0 MHz for Cl(3)SnMn(CO)(5) to 35.0 MHz for CH(3)Mn(CO)(5). The breadth of the solid-state (55)Mn NMR spectra of the pentacarbonyl halides is dominated by the CSA at all three applied magnetic fields. DFT calculations of the Mn magnetic shielding tensors reproduce the experimental trends and the magnitude of the CSA is qualitatively rationalized using a molecular orbital, MO, interpretation based on Ramsey's theory of magnetic shielding. In addition to the energy differences between symmetry-appropriate occupied and virtual MOs, the d-character of the Mn MOs is important for determining the paramagnetic shielding contribution to the principal components of the magnetic shielding tensor.     

Unusual structural types in manganese cluster chemistry from the use of N,N,N',N'-tetrakis(2-hydroxyethyl)ethylenediamine: Mn8, Mn12, and Mn20 Clusters

The syntheses, crystal structures, and magnetochemical characterization are reported for three new mixed-valent Mn clusters [Mn(8)O(3)(OH)(OMe)(O(2)CPh)7(edte)(edteH(2))](2)CPh) (1), [Mn(12)O(4)(OH)(2)(edte)(4)C(l6)(H(2)O)(2)] (2), and [Mn(20)O(8)(OH)(4)(O(2)CMe)(6)(edte)(6)](ClO(4))(2) (3) (edteH(4) = (HOCH(2)CH(2))(2)NCH(2)CH(2)N(CH(2)CH(2)OH)(2) = N,N,N',N'-tetrakis(2-hydroxyethyl)ethylenediamine). The reaction of edteH(4) with Mn(O(2)CPh)(2), MnCl(2), or Mn(O(2)CMe)(2) gives 1, 2, and 3, respectively, which all possess unprecedented core topologies. The core of 1 comprises two edge-sharing [Mn(4)O(4)] cubanes connected to an additional Mn ion by a micro(3)-OH- ion and two alkoxide arms of edteH(22-). The core of 2 consists of a [Mn(12)(micro(4-)O)(4)](24+) unit with S4 symmetry. The core of 3 consists of six fused [Mn(4)O(4)] cubanes in a 3 x 2 arrangement and linked to three additional Mn atoms at both ends. Variable-temperature, solid-state dc and ac magnetization (M) studies were carried out on complexes 1-3 in the 5.0-300 K range. Fitting of the obtained M/Nmicro(B) vs H/T data by matrix diagonalization and including only axial zero-field splitting (ZFS) gave ground-state spin (S) and axial ZFS parameter (D) of S = 8, D = -0.30 cm-1 for 1, S = 7, D = -0.16 cm-1 for 2, and S = 8, D = -0.16 cm-1 for 3. The combined work demonstrates that four hydroxyethyl arms on an ethylenediamine backbone can generate novel Mn structural types not accessible with other alcohol-based ligands.     

EPR-ENDOR characterization of (17O, 1H, 2H) water in manganese catalase and its relevance to the oxygen-evolving complex of photosystem II

The synthesis of efficient water-oxidation catalysts demands insight into the only known, naturally occurring water-oxidation catalyst, the oxygen-evolving complex (OEC) of photosystem II (PSII). Understanding the water oxidation mechanism requires knowledge of where and when substrate water binds to the OEC. Mn catalase in its Mn(III)-Mn(IV) state is a protein model of the OEC's S(2) state. From (17)O-labeled water exchanged into the di-μ-oxo di-Mn(III,IV) coordination sphere of Mn catalase, CW Q-band ENDOR spectroscopy revealed two distinctly different (17)O signals incorporated in distinctly different time regimes. First, a signal appearing after 2 h of (17)O exchange was detected with a 13.0 MHz hyperfine coupling. From similarity in the time scale of isotope incorporation and in the (17)O μ-oxo hyperfine coupling of the di-μ-oxo di-Mn(III,IV) bipyridine model (Usov, O. M.; Grigoryants, V. M.; Tagore, R.; Brudvig, G. W.; Scholes, C. P. J. Am. Chem. Soc. 2007, 129, 11886-11887), this signal was assigned to μ-oxo oxygen. EPR line broadening was obvious from this (17)O μ-oxo species. Earlier exchange proceeded on the minute or faster time scale into a non-μ-oxo position, from which (17)O ENDOR showed a smaller 3.8 MHz hyperfine coupling and possible quadrupole splittings, indicating a terminal water of Mn(III). Exchangeable proton/deuteron hyperfine couplings, consistent with terminal water ligation to Mn(III), also appeared. Q-band CW ENDOR from the S(2) state of the OEC was obtained following multihour (17)O exchange, which showed a (17)O hyperfine signal with a 11 MHz hyperfine coupling, tentatively assigned as μ-oxo-(17)O by resemblance to the μ-oxo signals from Mn catalase and the di-μ-oxo di-Mn(III,IV) bipyridine model.     

Effect of Ca2+/Sr2+ substitution on the electronic structure of the oxygen-evolving complex of photosystem II: a combined multifrequency EPR, 55Mn-ENDOR, and DFT study of the S2 state

The electronic structures of the native Mn(4)O(x)Ca cluster and the biosynthetically substituted Mn(4)O(x)Sr cluster of the oxygen evolving complex (OEC) of photosystem II (PSII) core complexes isolated from Thermosynechococcus elongatus, poised in the S(2) state, were studied by X- and Q-band CW-EPR and by pulsed Q-band (55)Mn-ENDOR spectroscopy. Both wild type and tyrosine D less mutants grown photoautotrophically in either CaCl(2) or SrCl(2) containing media were measured. The obtained CW-EPR spectra of the S(2) state displayed the characteristic, clearly noticeable differences in the hyperfine pattern of the multiline EPR signal [Boussac et al. J. Biol. Chem.2004, 279, 22809-22819]. In sharp contrast, the manganese ((55)Mn) ENDOR spectra of the Ca and Sr forms of the OEC were remarkably similar. Multifrequency simulations of the X- and Q-band CW-EPR and (55)Mn-pulsed ENDOR spectra using the Spin Hamiltonian formalism were performed to investigate this surprising result. It is shown that (i) all four manganese ions contribute to the (55)Mn-ENDOR spectra; (ii) only small changes are seen in the fitted isotropic hyperfine values for the Ca(2+) and Sr(2+) containing OEC, suggesting that there is no change in the overall spin distribution (electronic coupling scheme) upon Ca(2+)/Sr(2+) substitution; (iii) the changes in the CW-EPR hyperfine pattern can be explained by a small decrease in the anisotropy of at least two hyperfine tensors. It is proposed that modifications at the Ca(2+) site may modulate the fine structure tensor of the Mn(III) ion. DFT calculations support the above conclusions. Our data analysis also provides strong support for the notion that in the S(2) state the coordination of the Mn(III) ion is square-pyramidal (5-coordinate) or octahedral (6-coordinate) with tetragonal elongation. In addition, it is shown that only one of the currently published OEC models, the Siegbahn structure [Siegbahn, P. E. M. Acc. Chem. Res.2009, 42, 1871-1880, Pantazis, D. A. et al. Phys. Chem. Chem. Phys.2009, 11, 6788-6798], is consistent with all data presented here. These results provide important information for the structure of the OEC and the water-splitting mechanism. In particular, the 5-coordinate Mn(III) is a potential site for substrate 'water' (H(2)O, OH(-)) binding. Its location within the cuboidal structural unit, as opposed to the external 'dangler' position, may have important consequences for the mechanism of O-O bond formation.     

The role of the Mg2+ cation in ATPsynthase studied by electron paramagnetic resonance using VO2+ and Mn2+ paramagnetic probes

The electron paramagnetic resonance (EPR), electron spin echo envelope modulation (ESEEM) and hyperfine sublevel correlation (HYSCORE) spectra of Mg2+-depleted chloroplast F1-ATPase substituted with stoichiometric VO2+ are reported. The ESEEM and HYSCORE spectra of the complex are dominated by the hyperfine and quadrupole interactions between the VO2+ paramagnet and two different nitrogen ligands with isotropic hyperfine couplings /A1/ = 4.11 MHz and /A2/ = 6.46 MHz and nuclear quadrupole couplings e2qQ1 approximately 3.89-4.49 MHz and e2qQ2 approximately 1.91-2.20 MHz, respectively. Aminoacid functional groups compatible with these magnetic couplings include a histidine imidazole, the epsilon-NH2 of a lysine residue, and the guanidinium group of an arginine. Consistent with this interpretation, very characteristic correlations are detected in the HYSCORE spectra between the 14N deltaM1 = 2 transitions in the negative quadrant, and also between some of the deltaM1 = 1 transitions in the positive quadrant. The interaction of the substrate and product ADP and ATP nucleotides with the enzyme has been studied in protein complexes where Mg2+ is substituted for Mn2+. Stoichiometric complexes of Mn x ADP and Mn x ATP with the whole enzyme show distinct and specific hyperfine couplings with the 31P atoms of the bonding phosphates in the HYSCORE (ADP, A(31Pbeta) = 5.20 MHz: ATP, A(31Pbeta) = 4.60 MHz and A(31Pgamma) = 5.90 MHz) demonstrating the role of the enzyme active site in positioning the di- or triphosphate chain of the nucleotide for efficient catalysis. When the complexes are formed with the isolated alpha or beta subunits of the enzyme, the HYSCORE spectra are substantially modified, suggesting that in these cases the nucleotide binding site is only partially structured.     

Proton positions in the Mn(2+) binding site of concanavalin A as determined by single-crystal high-field ENDOR spectroscopy

High-field (95 GHz) pulsed EPR and electron-nuclear double resonance (ENDOR) techniques have been used for the first time to determine coordinates of ligand protons of a high-spin metal center in a protein single crystal. The protein concanavalin A contains a Mn(2+) ion which is coordinated to two water molecules, a histidine residue, and three carboxylates. Single crystals of concanavalin A were grown in H(2)O and in D(2)O to distinguish the exchangeable water protons from the nonexchangeable protons of the imidazole group. Distinct EPR transitions were selected by performing the ENDOR measurements at different magnetic fields within the EPR spectrum. This selection, combined with the large thermal polarization achieved at 4.5 K and a magnetic field of approximately 3.4 T allowed us to assign the ENDOR signals to their respective M(S) manifolds, thus providing the signs of the hyperfine couplings. Rotation patterns were acquired in the ac and ab crystallographic planes. Two distinct crystallographic sites were identified in each plane, and the hyperfine tensors of two of the imidazole protons and the four water protons were determined by simulations of the rotation patterns. All protons have axially symmetric hyperfine tensors and, by applying the point-dipole approximation, the positions of the various protons relative to the Mn(2+) ion were determined. Likewise, the water protons involved in H-bonding to neighboring residues were identified using the published, ultrahigh-resolution X-ray crystallographic coordinates of the protein (Deacon et al. J. Chem. Soc., Faraday Trans. 1997, 93(24), 4305-4312).     

The rotational spectrum and theoretical study of a dinuclear complex, MnRe(CO)(10)

The first rotational spectrum of a dinuclear complex, MnRe(CO)(10), has been obtained using a high-resolution pulsed beam microwave spectrometer. Sixty-four hyperfine components of the J=11-->J(')=12 and J=12-->J(')=13 rotational transitions were measured for two rhenium isotopomers. The B values obtained from the experiment are B=200.36871(18) MHz for the (187)Re isotopomer and B=200.5561(10) MHz for the (185)Re isotopomer. The measured rotational constants are in reasonably good agreement with the B values calculated from the x-ray diffraction structural data, and from theoretical calculations. The gas-phase Mn-Re bond distance is approximately 2.99 A, and the calculated value is only slightly longer. The experimental quadrupole coupling constant for the manganese atom is eQq(aa) ((55)Mn)=-16.52(5) MHz, and the corresponding quadrupole coupling constants for the two rhenium isotopomers are eQq(aa) ((187)Re)=370.4(4) MHz and eQq(aa) ((185)Re)=390.9(6) MHz. The quadrupole coupling constants were also determined from a variety of theoretical calculations, with very large Gaussian orbital bases. The best estimates, at a nonrelativistic level, are eQq(aa) ((55)Mn)=0.68 MHz and eQq(aa) ((187)Re)=327.6 MHz with a 874 GTO basis set, but the results are very basis set dependent, especially the sign of the Mn quadrupole coupling. Very slight bending of angles MnC(eq)O(eq) and ReC(eq)O(eq) angles is found in the calculations.     

EPR and ENDOR analysis of Fe3+ impurity centers in fluoroelpasolite lattices

Fe(3+) ions in hexagonal and cubic fluoroelpasolite crystals (A(1)(2)B(I)M(III)F(6)) have been investigated in a combined Electron Paramagnetic Resonance (EPR) and Electron Nuclear Double Resonance (ENDOR) study. A detailed analysis of the ENDOR spectra for the nearest (19)F and (23)Na shells in X (9.5 GHz) and Q band (34 GHz) allowed the complex EPR spectra to be disentangled and to determine the spin Hamiltonian parameters for the various S = 5/2 Fe(3+) centres. W-band (95 GHz) EPR measurements as a function of temperature were performed to provide unambiguous evidence about the absolute signs of the Zero Field Splitting (ZFS) and SuperHyperFine (SHF) parameters for Fe(3+) in Cs(2)NaAlF(6) as already determined from the ENDOR work. It could be concluded that all principal (19)F hyperfine values were positive, in agreement with earlier assignments in the literature for related systems. A comparative analysis of the (19)F SHF data for Fe(3+) at a perfectly octahedral site in the cubic crystal, and at two slightly trigonally distorted environments in the hexagonal crystals, indicates that the metal-to-ligand distance changes upon doping. The obtained set of parameters concerning one defect in various analogous environments can furthermore be used to test different methods of theoretical calculations for ZFS and SHF values.     

Theoretical investigation of zero field splitting parameters for Mn2+ centres in L-asparagine monohydrate

Zero-field splitting (ZFS) parameters D and E are calculated using the point-charge model (PCM) and superposition model (SPM) for Mn(2+) centre in L-asparagine monohydrate (LAM) single crystal. The calculated ZFS parameters obtained using these two models are compared with the experimental values for interstitial site of Mn(2+). The SPM and PCM give ZFS parameters similar to those of experimental ones. This supports the notion that the impurity ion occupies interstitial site in LAM.     

Density Functional Calculations of 55Mn, 14N and 13C Electron Paramagnetic Resonance Parameters Support an Energetically Feasible Model System for the S2 State of the Oxygen‐Evolving Complex of Photosystem II

Metal and ligand hyperfine couplings of a previously suggested, energetically feasible Mn₄Ca model cluster ( SG2009^?1 ) for the S₂ state of the oxygen‐evolving complex (OEC) of photosystem II (PSII) have been studied by broken‐symmetry density functional methods and compared with other suggested structural and spectroscopic models. This was carried out explicitly for different spin‐coupling patterns of the S=1/2 ground state of the Mn^III(Mn^IV)₃ cluster. By applying spin‐projection techniques and a scaling of the manganese hyperfine couplings, computation of the hyperfine and nuclear quadrupole coupling parameters allows a direct evaluation of the proposed models in comparison with data obtained from the simulation of EPR, ENDOR, and ESEEM spectra. The computation of ⁵⁵Mn hyperfine couplings (HFCs) for SG2009^?1 gives excellent agreement with experiment. However, at the current level of spin projection, the ⁵⁵Mn HFCs do not appear sufficiently accurate to distinguish between different structural models. Yet, of all the models studied, SG2009^?1 is the only one with the Mn^III site at the Mn_C center, which is coordinated by histidine (D1‐His332). The computed histidine ¹⁴N HFC anisotropy for SG2009^?1 gives much better agreement with ESEEM data than the other models, in which Mn_C is an Mn^IV site, thus supporting the validity of the model. The ¹³C HFCs of various carboxylates have been compared with ¹³C ENDOR data for PSII preparations with ¹³C‐labelled alanine.     

Carbonate complexation of Mn2+ in the aqueous phase: redox behavior and ligand binding modes by electrochemistry and EPR spectroscopy

The chemical speciation of Mn2+ within cells is critical for its transport, availability, and redox properties. Herein we investigate the redox behavior and complexation equilibria of Mn2+ in aqueous solutions of bicarbonate by voltammetry and electron paramagnetic resonance (EPR) spectroscopy and discuss the implications for the uptake of Mn2+ by mangano-cluster enzymes such as photosystem II (PSII). Both the electrochemical reduction of Mn2+ to Mn0 at an Hg electrode and EPR (in the absence of a polarizing electrode) revealed the formation of 1:1 and 1:2 Mn-(bi)carbonate complexes as a function of Mn2+ and bicarbonate concentrations. Pulsed EPR spectroscopy, including ENDOR, ESEEM, and 2D-HYSCORE, were used to probe the hyperfine couplings to 1H and 13C nuclei of the ligand(s) bound to Mn2+. For the 1:2 complex, the complete 13C hyperfine tensor for one of the (bi)carbonate ligands was determined and it was established that this ligand coordinates to Mn2+ in bidentate mode with a 13C-Mn distance of 2.85 +/- 0.1 angstroms. The second (bi)carbonate ligand in the 1:2 complex coordinates possibly in monodentate mode, which is structurally less defined, and its 13C signal is broad and unobservable. 1H ENDOR reveals that 1-2 water ligands are lost upon binding of one bicarbonate ion in the 1:1 complex while 3-4 water ligands are lost upon forming the 1:2 complex. Thus, we deduce that the dominant species above 0.1 M bicarbonate concentration is the 1:2 complex, [Mn(CO3)(HCO3)(OH2)3]-.     

Carboxylate-free Mn(III)2Ln(III)2 (Ln = lanthanide) and Mn(III)2Y(III)2 complexes from the use of (2-hydroxymethyl)pyridine: analysis of spin frustration effects

The initial employment of 2-(hydroxymethyl)pyridine for the synthesis of Mn/Ln (Ln = lanthanide) and Mn/Y clusters, in the absence of an ancillary organic ligand, has afforded a family of tetranuclear [Mn(III)(2)M(III)(2)(OH)(2)(NO(3))(4)(hmp)(4)(H(2)O)(4)](NO(3))(2) (M = Dy, 1; Tb, 2; Gd, 3; Y; 4) anionic compounds. 1-4 possess a planar butterfly (or rhombus) core and are rare examples of carboxylate-free Mn/Ln and Mn/Y clusters. Variable-temperature dc and ac studies established that 1 and 2, which contain highly anisotropic Ln(III) atoms, exhibit slow relaxation of their magnetization vector. Fitting of the obtained magnetization (M) versus field (H) and temperature (T) data for 3 by matrix diagonalization and including only axial anisotropy (zero-field splitting, ZFS) showed the ground state to be S = 3. Complex 4 has an S = 0 ground state. Fitting of the magnetic susceptibility data collected in the 5-300 K range for 3 and 4 to the appropriate van Vleck equations revealed, as expected, extremely weak antiferromagnetic interactions between the paramagnetic ions; for 3, J(1) = -0.16(2) cm(-1) and J(2) = -0.12(1) cm(-1) for the Mn(III)···Mn(III) and Mn(III)···Gd(III) interactions, respectively. The S = 3 ground state of 3 has been rationalized on the basis of the spin frustration pattern in the molecule. For 4, J = -0.75(3) cm(-1) for the Mn(III)···Mn(III) interaction. Spin frustration effects in 3 have been quantitatively analyzed for all possible combinations of sign of J(1) and J(2).     

17O ENDOR detection of a solvent-derived Ni-(OH(x))-Fe bridge that is lost upon activation of the hydrogenase from Desulfovibrio gigas.

Crystallographic studies of the hydrogenases (Hases) from Desulfovibrio gigas (Dg) and Desulfovibrio vulgaris Miyazaki (DvM) have revealed heterodinuclear nickel-iron active centers in both enzymes. The structures, which represent the as-isolated (unready) Ni-A (S = (1)/(2)) enzyme state, disclose a nonprotein ligand (labeled as X) bridging the two metals. The bridging atom was suggested to be an oxygenic (O(2)(-) or OH(-)) species in Dg Hase and an inorganic sulfide in DvM Hase. To determine the nature and chemical characteristics of the Ni-X-Fe bridging ligand in Dg Hase, we have performed 35 GHz CW (17)O ENDOR measurements on the Ni-A form of the enzyme, exchanged into H(2)(17)O, on the active Ni-C (S = (1)/(2)) form prepared by H(2)-reduction of Ni-A in H(2)(17)O, and also on Ni-A formed by reoxidation of Ni-C in H(2)(17)O. In the native state of the protein (Ni-A), the bridging ligand does not exchange with the H(2)(17)O solvent. However, after a reduction/reoxidation cycle (Ni-A --> Ni-C --> Ni-A), an (17)O label is introduced at the active site, as seen by ENDOR. Detailed analysis of a 2-D field-frequency plot of ENDOR spectra taken across the EPR envelope of Ni-A((17)O) shows that the incorporated (17)O has a roughly axial hyperfine tensor, A((17)O) approximately [5, 7, 20] MHz, discloses its orientation relative to the g tensor, and also yields an estimate of the quadrupole tensor. The substantial isotropic component (a(iso)((17)O) approximately 11 MHz) of the hyperfine interaction indicates that a solvent-derived (17)O is indeed a ligand to Ni and thus that the bridging ligand X in the Ni-A state of Dg Hase is indeed an oxygenic (O(2)(-) or OH(-)) species; comparison with earlier EPR results by others indicates that the same holds for Ni-B. The small (57)Fe hyperfine coupling seen previously for Ni-A (A((57)Fe) approximately 0.9 MHz) is now shown to persist in Ni-C, A((57)Fe) approximately 0.8 MHz. However, the (17)O signal is lost upon reductive activation to the Ni-C state; reoxidation to Ni-A leads to the reappearance of the signal. Consideration of the electronic structure of the EPR-active states of the dinuclear center leads us to suggest that the oxygenic bridge in Ni-A(B) is lost in Ni-C and is re-formed from solvent upon reoxidation to Ni-A. This implies that the reductive activation to Ni-C opens Ni/Fe coordination sites which may play a central role in the enzyme's activity.