Michael P. Hendrich

Reversible Switching of Magnetism in Thiolate-Protected Au25 Superatoms

We report reversible switching of paramagnetism in a well-defined gold nanoparticle system consisting of atomically monodisperse nanoparticles containing 25 gold atoms protected by 18 thiolates [abbreviated as Au(25)(SR)(18)]. The magnetism in these nanoparticles can be switched on or off by precisely controlling the charge state of the nanoparticle, that is, the magnetic state of the Au(25)(SR)(18) nanoparticles is charge-neutral while the nonmagnetic state is an anionic form of the particle. Electron paramagnetic resonance (EPR) spectroscopy measurements establish that the magnetic state of the Au(25)(SR)(18) nanoparticles possess one unpaired spin per particle. EPR studies also imply an unusual electronic structure of the Au(25)(SR)(18) nanoparticle. Density functional theory calculations coupled with the experiments successfully explain the origin of the observed magnetism in a Au(25)(SR)(18) nanoparticle as arising from one unpaired spin having distinct P-like character and delocalized among the icosahedral Au(13) core of the particle in the highest occupied molecular orbital. The results suggest that the Au(25)(SR)(18) nanoparticles are best considered as ligand-protected superatoms.

Formation, Structure, and EPR Detection of a High Spin FeIV—Oxo Species Derived from Either an FeIII—Oxo or FeIII—OH Complex

High spin oxoiron(IV) complexes have been proposed to be a key intermediate in numerous nonheme metalloenzymes. The successful detection of similar complexes has been reported for only two synthetic systems. A new synthetic high spin oxoiron(IV) complex is now reported that can be prepared from a well-characterized oxoiron(III) species. This new oxoiron(IV) complex can also be prepared from a hydroxoiron(III) species via a proton-coupled electron transfer process--a first in synthetic chemistry. The oxoiron(IV) complex has been characterized with a variety of spectroscopic methods: FTIR studies showed a feature associated with the Fe-O bond at nu(Fe(16)O) = 798 cm(-1) that shifted to 765 cm(-1) in the (18)O complex; Mossbauer experiments show a signal with an delta = 0.02 mm/s and |DeltaE(Q)| = 0.43 mm/s, electronic parameters consistent with an Fe(IV) center, and optical spectra had visible bands at lambda(max) = 440 (epsilon(M) = 3100), 550 (epsilon(M) = 1900), and 808 (epsilon(M) = 280) nm. In addition, the oxoiron(IV) complex gave the first observable EPR features in the parallel-mode EPR spectrum with g-values at 8.19 and 4.06. A simulation for an S = 2 species with D = 4.0(5) cm(-1), E/D = 0.03, sigma(E/D) = 0.014, and g(z) = 2.04 generates a fit that accurately predicted the intensity, line shape, and position of the observed signals. These results showed that EPR spectroscopy can be a useful method for determining the properties of high spin oxoiron(IV) complexes. The oxoiron(IV) complex was crystallized at -35 degrees C, and its structure was determined by X-ray diffraction methods. The complex has a trigonal bipyramidal coordination geometry with the Fe-O unit positioned within a hydrogen bonding cavity. The Fe(IV)-O unit bond length is 1.680(1) A, which is the longest distance yet reported for a monomeric oxoiron(IV) complex.

Integer-spin electron paramagnetic resonance of iron proteins

A quantitative interpretation is presented for EPR spectra from integer-spin metal centers having large zero-field splittings. Integer-spin, or non-Kramers, centers are common in metalloproteins and many give EPR signals, but a quantitative understanding has been lacking until now. Heterogeneity of the metals local environment will result in a significant spread in zero-field splittings and in broadened EPR signals. Using the spin Hamiltonian Hs = S.D.S + beta S.g.B and some simple assumptions about the nature of the zero-field parameter distributions, a lineshape model was devised which allows accurate simulation of single crystal and frozen solution spectra. The model was tested on single crystals of magnetically dilute ferrous fluosilicate. Data and analyses from proteins and active-site models are presented with the microwave field B1 either parallel or perpendicular to B. Quantitative agreement of observed and predicted signal intensities is found for the two B1 orientations. Methods of spin quantitation are given and are shown to predict an unknown concentration relative to a standard with known concentration. The fact that the standard may be either a non-Kramers or a Kramers center is further proof of the models validity. The magnitude of the splitting in zero magnetic field is of critical importance; it affects not only the chance of signal observation, but also the quantitation accuracy. Experiments taken at microwave frequencies of 9 and 35 GHz demonstrate the need for high-frequency data as only a fraction of the molecules give signals at 9 GHz.

On the feasibility of N2 fixation via a single-site FeI/FeIV cycle: Spectroscopic studies of FeI(N2)FeI, FeIV N, and related species

The electronic properties of an unusually redox-rich iron system, [PhBPR3]FeNx (where [PhBPR3] is [PhB(CH2PR2)3]−), are explored by Mössbauer, EPR, magnetization, and density-functional methods to gain a detailed picture regarding their oxidation states and electronic structures. The complexes of primary interest in this article are the two terminal iron(IV) nitride species, [PhBPiPr3]FeN (3a) and [PhBPCH2Cy3]FeN (3b), and the formally diiron(I) bridged-Fe(μ-N2)Fe species, {[PhBPiPr3]Fe}2(μ-N2) (4). Complex 4 is chemically related to 3a via a spontaneous nitride coupling reaction. The diamagnetic iron(IV) nitrides 3a and 3b exhibit unique electronic environments that are reflected in their unusual Mössbauer parameters, including quadrupole-splitting values of 6.01(1) mm/s and isomer shift values of −0.34(1) mm/s. The data for 4 suggest that this complex can be described by a weak ferromagnetic interaction (J/D < 1) between two iron(I) centers. For comparison, four other relevant complexes also are characterized: a diamagnetic iron(IV) trihydride [PhBPiPr3]Fe(H)3(PMe3) (5), an S = 3/2 iron(I) phosphine adduct [PhBPiPr3]FePMe3 (6), and the S = 2 iron(II) precursors to 3a, [PhBPiPr3]FeCl and [PhBPiPr3]Fe-2,3:5,6-dibenzo-7-aza bicyclo[2.2.1]hepta-2,5-diene (dbabh). The electronic properties of these respective complexes also have been explored by density-functional methods to help corroborate our spectral assignments and to probe their electronic structures further.

Trapping and spectroscopic characterization of an FeIII-superoxo intermediate from a nonheme mononuclear iron-containing enzyme

intermediates are well known in heme enzymes, but none have been characterized in the nonheme mononuclear FeII enzyme family. Many steps in the O2 activation and reaction cycle of FeII-containing homoprotocatechuate 2,3-dioxygenase are made detectable by using the alternative substrate 4-nitrocatechol (4NC) and mutation of the active site His200 to Asn (H200N). Here, the first intermediate (Int-1) observed after adding O2 to the H200N-4NC complex is trapped and characterized using EPR and Mössbauer (MB) spectroscopies. Int-1 is a high-spin (S1 = 5/2) FeIII antiferromagnetically (AF) coupled to an S2 = 1/2 radical (J ≈ 6 cm-1 in ). It exhibits parallel-mode EPR signals at g = 8.17 from the S = 2 multiplet, and g = 8.8 and 11.6 from the S = 3 multiplet. These signals are broadened significantly by hyperfine interactions (A17O ≈ 180 MHz). Thus, Int-1 is an AF-coupled species. The experimental observations are supported by density functional theory calculations that show nearly complete transfer of spin density to the bound O2. Int-1 decays to form a second intermediate (Int-2). MB spectra show that it is also an AF-coupled FeIII-radical complex. Int-2 exhibits an EPR signal at g = 8.05 arising from an S = 2 state. The signal is only slightly broadened by (< 3% spin delocalization), suggesting that Int-2 is a peroxo-FeIII-4NC semiquinone radical species. Our results demonstrate facile electron transfer between FeII, O2, and the organic ligand, thereby supporting the proposed wild-type enzyme mechanism.

Formation of a Room Temperature Stable Fe V (O) Complex: Reactivity Toward Unactivated C−H Bonds

An Fe(V)(O) complex has been synthesized from equimolar solutions of (Et4N)2[Fe(III)(Cl)(biuret-amide)] and mCPBA in CH3CN at room temperature. The Fe(V)(O) complex has been characterized by UV-vis, EPR, Mössbauer, and HRMS and shown to be capable of oxidizing a series of alkanes having C-H bond dissociation energies ranging from 99.3 kcal mol(-1) (cyclohexane) to 84.5 kcal mol(-1) (cumene). Linearity in the Bell-Evans-Polayni graph and the finding of a large kinetic isotope effect suggest that hydrogen abstraction is engaged the rate-determining step.

Structure-Function, Stability, and Chemical Modification of the Cyanobacterial Cytochrome b6f Complex from Nostoc sp. PCC 7120

The crystal structure of the cyanobacterial cytochrome b6f complex has previously been solved to 3.0-Å resolution using the thermophilic Mastigocladus laminosus whose genome has not been sequenced. Several unicellular cyanobacteria, whose genomes have been sequenced and are tractable for mutagenesis, do not yield b6f complex in an intact dimeric state with significant electron transport activity. The genome of Nostoc sp. PCC 7120 has been sequenced and is closer phylogenetically to M. laminosus than are unicellular cyanobacteria. The amino acid sequences of the large core subunits and four small peripheral subunits of Nostoc are 88 and 80% identical to those in the M. laminosus b6f complex. Purified b6f complex from Nostoc has a stable dimeric structure, eight subunits with masses similar to those of M. laminosus, and comparable electron transport activity. The crystal structure of the native b6f complex, determined to a resolution of 3.0Å (PDB id: 2ZT9), is almost identical to that of M. laminosus. Two unique aspects of the Nostoc complex are: (i) a dominant conformation of heme bp that is rotated 180° about the α- and γ-meso carbon axis relative to the orientation in the M. laminosus complex and (ii) acetylation of the Rieske iron-sulfur protein (PetC) at the N terminus, a post-translational modification unprecedented in cyanobacterial membrane and electron transport proteins, and in polypeptides of cytochrome bc complexes from any source. The high spin electronic character of the unique heme cn is similar to that previously found in the b6f complex from other sources.

Electron Paramagnetic Resonance Detection of Intermediates in the Enzymatic Cycle of an Extradiol Dioxygenase

Extradiol catecholic dioxygenases catalyze the cleavage of the aromatic ring of the substrate with incorporation of both oxygen atoms from O2. These enzymes are important in nature for the recovery of large amounts of carbon from aromatic compounds. The catalytic site contains either Fe or Mn coordinated by a facial triad of two His and one Glu or Asp residues. Previous studies have shown that Fe(II) and Mn(II) can be interchanged in enzymes from different organisms to catalyze similar substrate reactions. In combination, quantitative electron paramagnetic resonance spectroscopy and rapid freeze-quench experiments allow us to follow the concentrations of four different Mn species, including key metal intermediates in the catalytic cycle, as the enzyme turns over its natural substrate. Two intermediates are observed: a Mn(III)-radical species which is either Mn-superoxide or Mn-substrate radical, and a unique Mn(II) species which is involved in the rate-limiting step of the cycle and may be Mn-alkylperoxo.

EPR and Mössbauer Spectroscopy of Intact Mitochondria Isolated from Yah1p-Depleted Saccharomyces cerevisiae†

Yah1p, an [Fe 2S 2]-containing ferredoxin located in the matrix of Saccharomyces cerevisiae mitochondria, functions in the synthesis of Fe/S clusters and heme a prosthetic groups. EPR, Mossbauer spectroscopy, and electron microscopy were used to characterize the Fe that accumulates in Yah1p-depleted isolated intact mitochondria. Gal- YAH1 cells were grown in standard rich media (YPD and YPGal) under O 2 or argon atmospheres. Mitochondria were isolated anaerobically, then prepared in the as-isolated redox state, the dithionite-treated state, and the O 2-treated state. The absence of strong EPR signals from Fe/S clusters when Yah1p was depleted confirms that Yah1p is required in Fe/S cluster assembly. Yah1p-depleted mitochondria, grown with O 2 bubbling through the media, accumulated excess Fe (up to 10 mM) that was present as 2-4 nm diameter ferric nanoparticles, similar to those observed in mitochondria from yfh1Delta cells. These particles yielded a broad isotropic EPR signal centered around g = 2, characteristic of superparamagnetic relaxation. Treatment with dithionite caused Fe (3+) ions of the nanoparticles to become reduced and largely exported from the mitochondria. Fe did not accumulate in mitochondria isolated from cells grown under Ar; a significant portion of the Fe in these organelles was in the high-spin Fe (2+) state. This suggests that the O 2 used during growth of Gal- YAH1 cells is responsible, either directly or indirectly, for Fe accumulation and for oxidizing Fe (2+) --> Fe (3+) prior to aggregation. Models are proposed in which the accumulation of ferric nanoparticles is caused either by the absence of a ligand that prevents such precipitation in wild-type mitochondria or by a more oxidizing environment within the mitochondria of Yah1p-depleted cells exposed to O 2. The efficacy of reducing accumulated Fe along with chelating it should be considered as a strategy for its removal in diseases involving such accumulations.

Mössbauer and EPR study of the binuclear iron centre in purple acid phosphatase

Mössbauer spectra have been determined on 57Fe-enriched samples of both pink (reduced) and purple (oxidized) forms of pig allantoic acid phosphatase (EC 3.1.3.2), and EPR spectra on corresponding unenriched samples. The spectra show unambiguously that both forms of the enzyme contain two distinct, antiferromagnetically coupled, high-spin iron atoms: a ferrous-ferric ion pair in the pink, reduced form, and a pair of ferric ions in the purple, oxidized form.