Electron Nuclear Double Resonance Spectra Research Articles

Electron Spin Coherence and Electron Nuclear Double Resonance of Bi Donors in Natural Si

Donors in silicon hold considerable promise for emerging quantum technologies, due to their uniquely long electron spin coherence times. Bismuth donors in silicon differ from more widely studied group V donors, such as phosphorous, in several significant respects: They have the strongest binding energy (70.98meV), a large nuclear spin (I=9/2), and a strong hyperfine coupling constant (A=1475.4 MHz). These larger energy scales allow us to perform a detailed test of theoretical models describing the spectral diffusion mechanism that is known to govern the electron spin decoherence of P donors in natural silicon. We report the electron-nuclear double resonance spectra of the Bi donor, across the range 200MHz to 1.4GHz, and confirm that coherence transfer is possible between electron and nuclear spin degrees of freedom at these higher frequencies.

Single-crystal EPR and ENDOR study of an AlO center in prehnite: implications for aluminum-associated oxyradicals in layer silicates

Single-crystal electron paramagnetic resonance (EPR) spectra of gamma-ray-irradiated prehnite (Jeffrey mine, Quebec, Canada) measured at 298 and 160 K reveal an aluminum-associated oxygen hole center (Al—O − ). Spin Hamiltonian parameters g and A ( 27 Al) fitted from the 298 K spectra suggest that this Al—O − center represents hole trapping on an apical hydroxyl oxygen atom (after removal of the proton) coordinated to an octahedral Al 3+ ion ( i.e. , an [ • OAlO 4 (OH)] center from the [(OH)AlO 4 (OH)] precursor, where • denotes the unpaired spin). Pulsed electron nuclear double resonance (ENDOR) spectra measured at 25 K allow the identification and quantitative analysis of two sets of 27 Al hyperfine structures and five proton hyperfine structures, which are all consistent with the proposed structural model. Isothermal and isochronal annealing experiments show that this center is bleached out completely at 375 °C, but can be readily restored by gamma-ray irradiation, and exhibits second-order decay kinetics. These results from the Al—O − center in prehnite provide support for and new insights into Clozel et al . (1995)’s VI Al—O − — VI Al model for B-centers in kaolinite.

Identification of electron and hole traps in lithium tetraborate (Li2B4O7) crystals: Oxygen vacancies and lithium vacancies

Electron paramagnetic resonance (EPR) and electron-nuclear double resonance (ENDOR) are used to identify and characterize electrons trapped by oxygen vacancies and holes trapped by lithium vacancies in lithium tetraborate (Li2B4O7) crystals. Our study includes a crystal with the natural abundances of B10 and B11 and a crystal highly enriched with B10. The as-grown crystals contain isolated oxygen vacancies, lithium vacancies, and copper impurities, all in nonparamagnetic charge states. During an irradiation at 77 K with 60 kV x-rays, doubly ionized oxygen vacancies trap electrons while singly ionized lithium vacancies and monovalent copper impurities trap holes. The vacancies return to their preirradiation charge states when the temperature of the sample is increased to approximately 90 K. Hyperfine interactions with B10 and B11 nuclei, observed between 13 and 40 K in the radiation-induced EPR and ENDOR spectra, provide models for the two vacancy-related defects. The electron trapped by an oxygen vacancy is localized primarily on only one of the two neighboring boron ions while the hole stabilized by a lithium vacancy is localized on a neighboring oxygen ion with nearly equal interactions with the two boron ions adjacent to the oxygen ion.

Solvation of Small Disulfonate Anions in Water/Methanol Mixtures Characterized by High-Field Pulse Electron Nuclear Double Resonance and Molecular Dynamics Simulations

The solvation of Fremy's salt, the paramagnetic nitrosodisulfonate anion ON(SO(3)(-))(2), in binary solvent mixtures was investigated by means of pulse (Mims- and Davies-type) electron nuclear double resonance (ENDOR) spectroscopy and molecular dynamics (MD) simulations. (1)H and (2)H pulse ENDOR measurements were performed on small Fremy's salt radicals in isotope-substituted solvent mixtures of methanol and water in frozen solution. We were able to obtain well-resolved, orientation-selective, high-field/high-frequency pulse ENDOR spectra of methyl protons from the alcohol moiety and exchangeable protons from the alcohol-hydroxyl group and water. In the studied solvent systems (volume ratio v/v = 30:70, 50:50, 70:30), the solvation of 2.5 mM Fremy's salt by methyl protons was found to be almost identical. From the analysis of the dependence of pulse ENDOR spectra on the observer field position and spectral simulations, we obtained the principal components of the hyperfine coupling (hfc) tensor for each class of protons. The combination of Mims- and Davies-type pulse ENDOR measurements was necessary to obtain blind spot free information on hfc that spans a broad range of 0.25-6 MHz. Using the point-dipole approximation, the dipolar hfc component yields a prominent electron-nuclear distance of 3.5 A between Fremy's salt and methyl protons, which was found along the molecular z-axis (perpendicular to the approximate plane spanned by ON(S)(2)) of the probe molecule. Exchangeable protons were found to be distributed nearly isotropically, forming a hydrogen-bonded network around the sulfonate groups. The distribution of exchangeable and methyl protons found in MD simulations is in very good agreement with the pulse ENDOR results, and we find that solvation is dominated by an interplay of H-bond (electrostatic) interactions and steric properties. The elucidation of the microscopic solvation of a small probe molecule in binary solvent mixtures represents the first step for understanding the interactions in more complex biochemical systems. In particular, this includes the potential perturbation of the H-bond network due to the presence of a spin probe or other polar molecules.

Magnetic resonance study of H atoms in thin films ofH2at temperatures below 1 K

We present an experimental study of H atoms embedded in thin films of solid ${\text{H}}_{2}$ at temperatures below 1 K. ${\text{H}}_{2}$ films up to 50 nm thick were first grown as a result of slow recombination of atomic hydrogen gas on the sample cell walls. If the recombination occurred in three-body atomic collisions in the gas phase, small concentrations of atoms could be captured inside the films during the film deposition. As a second method of generating atomic populations inside the ${\text{H}}_{2}$ films, we used a direct dissociation by a low power rf discharge in the sample cell. With this latter method, we achieved record high atomic concentrations exceeding $2\ifmmode\times\else\texttimes\fi{}{10}^{19}\text{ }{\text{cm}}^{\ensuremath{-}3}$. The samples were characterized by means of magnetic resonance: electron spin resonance (ESR) and electron-nuclear double resonance (ENDOR) in a magnetic field of 4.6 T. We observed density-dependent broadening and shifts of the ESR lines due to the dipolar interactions, and resolved these effects for like and unlike atoms. Relaxation of the relative hyperfine populations was measured as a function of temperature for H in ${\text{H}}_{2}$ films grown on different substrates. For ${\text{H}}_{2}$ films on Mylar substrates, the relative equilibrium populations of the two lowest hyperfine states of H were found to deviate substantially from the prediction of Boltzmann statistics. We also found two narrow lines in the ENDOR spectra of H in ${\text{H}}_{2}$ films shifted to the red from the position for free atoms. This indicates two possible substitutional positions of the atoms in ${\text{H}}_{2}$ matrices, both characterized by very homogeneous crystalline fields.

Binding of histidine in the (Cys)3(His)1-coordinated [2Fe-2S] cluster of human mitoNEET.

Human mitoNEET is a homodimeric iron-sulfur protein located in the outer mitochondrial membrane with unknown function, but which is known to interact with thiazolidinedione diabetes drugs. Each monomer houses a [2Fe-2S] cluster with an unusual (Cys)(3)(His)(1) ligation. The His ligand is important for enabling cluster release and for tuning the redox potential. We use multifrequency (X-, Ka-, and Q-band) and multitechnique (continuous-wave, electron spin-echo envelope modulation (ESEEM), pulsed electron-nuclear double resonance (ENDOR), and hyperfine sublevel correlation (HYSCORE)) electron paramagnetic resonance spectroscopy to investigate the cluster in its paramagnetic reduced [Fe(2+)Fe(3+)] (S = 1/2) state. It has a rhombic g tensor (2.007, 1.937, 1.897) with an average g value of 1.947 that falls between those of Rieske-type and ferredoxin-type [2Fe-2S] clusters. Simulation and least-squares fitting of orientation-selective Ka- and Q-band ENDOR, 1D ESEEM, and HYSCORE spectra of (14)N and (15)N-labeled mitoNEET yield the principal values and orientations of both the hyperfine tensor ((14)N, A(iso) = -6.25 MHz, T = -0.94 MHz) and the quadrupolar tensor (e(2)Qq/h = -2.47 MHz, eta = 0.38) of the ligating histidine nitrogen N(delta). From these, we can infer the absolute g tensor orientation with respect to the cluster: The g(2) axis is close to perpendicular to the [2Fe-2S] plane, and g(1) and g(3) are in-plane, but skewed from the Fe-Fe and S-S axes. In X-band ENDOR and ESEEM spectra, a weakly coupled nitrogen is visible, most likely the N(epsilon) of the histidine in the protonated state. We find that the cluster is in a valence-localized state, where Fe(2+) is His-bound. The field-sweep spectra show evidence of intercluster dipolar coupling that can be simulated using an uncoupled spin model for each cluster (S(Fe(2+)) = 2, S(Fe(3+)) = 5/2). The parameters determined in this work can function as reporters on how the cluster structure is altered upon pH changes and drug binding.

Fluorine donors andTi3+ions inTiO2crystals

Electron paramagnetic resonance (EPR) and electron-nuclear double resonance (ENDOR) are used to identify and characterize neutral fluorine donors in ${\text{TiO}}_{2}$ crystals having the rutile structure. The fluorine ions substitute for oxygen and are present as unintentional impurities. Isolated singly ionized fluorine donors in an as-grown (fully oxidized) crystal convert to their neutral charge state during exposure to 442 nm laser light at 6 K. These donors return to the singly ionized charge state within a few seconds when the light is removed. In contrast, the neutral fluorine donors are observed at 6 K without photoexcitation after a crystal is reduced at $600\text{ }\ifmmode^\circ\else\textdegree\fi{}\text{C}$ in flowing nitrogen gas. The angular dependences of the EPR and ENDOR spectra provide a complete set of spin-Hamiltonian parameters (principal values are 1.9746, 1.9782, and 1.9430 for the $g$ matrix and $\ensuremath{-}0.23$, 0.47, and 5.15 MHz for the $^{19}\text{F}$ hyperfine matrix). These matrices suggest that the unpaired electron is localized primarily on one of the two equivalent neighboring substitutional titanium ions, i.e., the ground state of the neutral fluorine donor in rutile-structured ${\text{TiO}}_{2}$ is a ${\text{Ti}}^{3+}$ ion adjacent to a ${\text{F}}^{\ensuremath{-}}$ ion.

Investigation of the Stationary and Transient A(1) Radical in Trp --> Phe Mutants of Photosystem I.

Photosystem I (PS I) contains two symmetric branches of electron transfer cofactors. In both the A- and B-branches, the phylloquinone in the A1 site is π-stacked with a tryptophan residue and is H-bonded to the backbone nitrogen of a leucine residue. In this work, we use optical and electron paramagnetic resonance (EPR) spectroscopies to investigate cyanobacterial PS I complexes, where these tryptophan residues are changed to phenylalanine. The time-resolved optical data show that backward electron transfer from the terminal electron acceptors to P700·+ is affected in the A- and B-branch mutants, both at ambient and cryogenic temperatures. These results suggest that the quinones in both branches take part in electron transport at all temperatures. The electron-nuclear double resonance (ENDOR) spectra of the spin-correlated radical pair P700·+A1·− and the photoaccumulated radical anion A1·−, recorded at cryogenic temperature, allowed the identification of characteristic resonances belonging to protons of the methyl group, some of the ring protons and the proton hydrogen-bonded to phylloquinone in the wild type and both mutants. Significant changes in PS I isolated from the A-branch mutant are detected, while PS I isolated from the B-branch mutant shows the spectral characteristics of wild-type PS I. A possible short-lived B-branch radical pair cannot be detected by EPR due to the available time resolution; therefore, only the A-branch quinone is observed under conditions typically employed for EPR and ENDOR spectroscopies.

Formation of {[HIPTN(3)N]Mo(III)H}(-) by heterolytic cleavage of H(2) as established by EPR and ENDOR spectroscopy.

MoN(2) (Mo = [(HIPTNCH(2)CH(2))(3)N]Mo, where HIPT = 3,5-(2,4,6-i-Pr(3)C(6)H(2))(2)C(6)H(3)) is the first stage in the reduction of N(2) to NH(3) by Mo. Its reaction with dihydrogen in fluid solution yields "MoH(2)", a molybdenum-dihydrogen compound. In this report, we describe a comprehensive electron paramagnetic resonance (EPR) and (1/2)H/(14)N electron nuclear double resonance (ENDOR) study of the product of the reaction between MoN(2) and H(2) that is trapped in frozen solution, 1. EPR spectra of 1 show that it has a near-axial g tensor, g = [2.086, 1.961, 1.947], with dramatically reduced g anisotropy relative to MoN(2). Analysis of the g values reveal that this anion has the Mo(III), [d(xz), d(yz)](3) orbital configuration, as proposed for the parent MoN(2) complex, and that it undergoes a strong pseudo-Jahn-Teller (PJT) distortion. Simulations of the 2D 35 GHz (1)H ENDOR pattern comprised of spectra taken at multiple fields across the EPR envelope (2 K) show that 1 is the [MoH](-) anion. The 35 GHz Mims pulsed (2)H ENDOR spectra of 1 prepared with (2)H(2) show the corresponding (2)H(-) signal, with a substantial deuterium isotope effect in a(iso). Radiolytic reduction of a structural analogue, Mo(IV)H, at 77 K, confirms the assignment of 1. Analysis of the 2D (14)N ENDOR pattern for the ligand amine nitrogen further reveals the presence of a linear N(ax)-Mo-H(-) molecular axis that is parallel to the unique magnetic direction (g(1)). The ENDOR pattern of the three equatorial nitrogens is well-reproduced by a model in which the Mo-N(eq) plane has undergone a static, not dynamic, PJT distortion, leading to a range of hyperfine couplings for the three N(eq)'s. The finding of a nearly axial hyperfine coupling tensor for the terminal hydride bound Mo supports the earlier proposal that the two exchangeable hydrogenic species bound to the FeMo cofactor of the nitrogense turnover intermediate, which has accumulated four electrons/protons (E(4)), are hydrides that bridge two metal ions, not terminal hydrides.

The Electronic State of Flavoproteins: Investigations with Proton Electron–Nuclear Double Resonance

Electron-nuclear double resonance (ENDOR) spectroscopy provides useful information on hyperfine interactions between nuclear magnetic moments and the magnetic moment of an unpaired electron spin. Because the hyperfine coupling constant reacts quite sensitively to polarity changes in the direct vicinity of the nucleus under consideration, ENDOR spectroscopy can be favorably used for the detection of subtle protein-cofactor interactions. A number of pulsed ENDOR studies on flavoproteins have been published during the past few years; most of them were designed to characterize the flavin cofactor by means of its protonation state, or to detect individual protein-cofactor interactions. The aim of this study is to compare the pulsed ENDOR spectra from different flavoproteins in terms of variations of characteristic proton hyperfine values. The general concept is to observe limits of possible influences on the cofactor's electronic state by surrounding amino acids. Furthermore, we compare ENDOR data obtained from in vivo experiments with in vitro data to emphasize the potential of the method for gaining molecular information in complex media.

EPR and ENDOR Investigation of Rhodosemiquinone in Bacterial Reaction Centers Formed by B-Branch Electron Transfer

In photosynthetic bacteria, light-induced electron transfer takes place in a protein called the reaction center (RC) leading to the reduction of a bound ubiquinone molecule, Q(B), coupled with proton binding from solution. We used electron paramagnetic resonance (EPR) and electron-nuclear double resonance (ENDOR) to study the magnetic properties of the protonated semiquinone, an intermediate proposed to play a role in proton coupled electron transfer to Q(B). To stabilize the protonated semiquinone state, we used a ubiquinone derivative, rhodoquinone, which as a semiquinone is more easily protonated than ubisemiquinone. To reduce this low-potential quinone we used mutant RCs modified to directly reduce the quinone in the Q(B) site via B-branch electron transfer (Paddock et al. in Biochemistry 44:6920-6928, 2005). EPR and ENDOR signals were observed upon illumination of mutant RCs in the presence of rhodoquinone. The EPR signals had g values characteristic of rhodosemiquinone (g(x) = 2.0057, g(y) = 2.0048, g(z) ∼ 2.0018) at pH 9.5 and were changed at pH 4.5. The ENDOR spectrum showed couplings due to solvent exchangeable protons typical of hydrogen bonds similar to, but different from, those found for ubisemiquinone. This approach should be useful in future magnetic resonance studies of the protonated semiquinone.

Pulse EPR, ENDOR, and ELDOR Study of Anionic Flavin Radicals in Na+-Translocating NADH:Quinone Oxidoreductase

The Na+-translocating nicotinamide adenine dinucleotide (NADH):quinine oxidoreductase (Na+–NQR) is a component of respiratory chain of various bacteria and it generates a redox-driven transmembrane electrochemical Na+ potential. It contains four different flavin prosthetic groups, including two flavin mononucleotide (FMN) residues covalently bound to the subunits NqrB and NqrC. Na+–NQR from Vibrio harveyi was poised at different redox potentials to prepare two samples, containing either both FMNNqrB and FMNNqrC or only FMNNqrB in a paramagnetic state. These two samples were comparatively studied using pulse electron paramagnetic resonance (EPR), electron-nuclear double resonance (ENDOR), and electron-electron double resonance (ELDOR) spectroscopy. The echo-detected EPR spectra and electron spin relaxation properties were very similar for flavin radicals in both samples. The splitting of the outer peaks in the proton ENDOR spectra, assigned to the C(8α) methyl protons, allows to identify both radicals as anionic flavosemiquinones. The mean interspin distance of 20.7 A between these radicals was determined by pulse ELDOR experiment, which allows to estimate the edge-to-edge distance (re) between these flavin centers as: 11.7 A < re < 20.7 A. The direct electron transfer between FMNNqrB and FMNNqrC during the physiological turnover of the Na+–NQR complex is suggested.

Incorporation of 2,3-Disubstituted-1,4-Naphthoquinones into the A1 Binding Site of Photosystem I Studied by EPR and ENDOR Spectroscopy

Transient electron paramagnetic resonance and pulsed electron-nuclear double resonance (ENDOR) spectra of the state \( P_{700}^{ \cdot + } A_{1}^{ \cdot - } \) in photosystem I containing a series of non-native naphthoquinones (NQs) are presented. Previous studies have shown that quinones bind to the A1 site with only one of their carbonyl groups H-bonded to the protein and that the asymmetric H-bond produces an odd alternant distribution of the spin density within the quinone. It is known that the native phylloquinone binds with its methyl group meta and its phytyl tail ortho to the H-bonded carbonyl. Monosubstituted NQs with short alkyl chains have been found to bind preferentially with their alkyl side groups meta to the H-bonded carbonyl. The selectivity of the binding site toward methyl and short chain substituents is studied by incorporating disubstituted NQs that have a methyl group at the 2-position and a short chain at the 3-position of the quinone ring. The hyperfine couplings (hfcs) of the methyl group protons are sensitive to the spin density distribution on the quinone and are used to deduce the position of the methyl group relative to the H-bonded carbonyl. The measured methyl proton hfcs indicate that the disubstituted quinones bind exclusively with their methyl group in the meta position relative to the H-bonded carbonyl and no evidence for binding with the methyl group in the ortho position is found. The disubstituted quinones have also been chosen to study the effect of electron withdrawing substituents on the spin density distribution. When the short chain contains electronegative atoms such as sulfur or chlorine, the methyl proton hfcs of the quinone in the A1 binding site are found to be significantly larger than those of 2-methyl-1,4-naphthoquinone and phylloquinone in the same environment. Solution ENDOR measurements of the quinone radical anions in isopropanol and density functional theory (DFT) calculations in vacuo show that this increase in the hfcs is mostly intrinsic to the quinones due to the electron-withdrawing ability of the short chain and is not a result of differences in the binding to the protein. The DFT calculations suggest that the main reason for the increased methyl proton hfcs is delocalization of the singly occupied molecular orbital onto the side chain, which leads to an increase of the spin density on the neighboring carbon, which carries methyl group.

Room Temperature Solution ENDOR of some Super Stable Fluorocarbon Radicals

Room temperature fluorine electron nuclear double resonance (ENDOR) has been successfully observed for several superstable fluorocarbon radicals ·C(C2F4R)(i-C3F7)2 in solution. Three radicals were employed in which CF3, F, and O-c-C6F10SO3C2F5 were introduced as R, and all the hyperfine couplings (hfcs) obtained by ENDOR were assigned with the help of ESR simulation and ab initio MO calculation. In case of ·C(i-C3F7)3 large 13C and considerable β-fluorine couplings suggest the nonplaner arrangement for the central and three carbons at the β-position, in spite of the fact that all the methyl fluorine show the same hfc. Therefore, a rapid puckering motion along the C3 axis together with the methyl rotation should average the hfc’s of the 18 fluorine nuclei to give the same value. When one of the CF3 groups is substituted with an F nucleus, the five CF3 groups give two hfc values, suggesting some dynamics still exists for the molecular frame. When a large group, O-c-C6F10SO3C2F5, is substituted for CF3, all the five CF3 groups become nonequivalent and the ENDOR signal becomes intensive and sharp even at 290 K, indicating that the molecular frame becomes rigid. The relation between the ENDOR spectra of these systems and the intramolecular dynamics is discussed.

Spin‐coupling and hyperfine interaction of the nitrogen donors in 6H‐SiC

AbstractThe X‐, Q‐band field‐sweep electron spin echo (FS ESE) study of n‐type 6H‐SiC wafers shows besides the well known hyperfine (hf) triplet lines of14N on two quasi‐cubic (Nc1, Nc2) and hexagonal (Nh) sites additional triplet lines (Nx) of comparatively low intensity with ag‐tensor of about the average value of the Nc2and Nhspectrum at 4.2 K. The Nxtriplet has half of the hf splitting with respect to the nitrogen residing at cubic sites Nc2. Pulsed electron nuclear double resonance (ENDOR) measurements performed on the NxEPR triplet line demonstrate that there is an efficient spin‐coupling between Nhand Nxnitrogen related centers. The Nxtriplet lines were attributed to distant donor Nc–Nhpairs between nitrogen atoms residing at quasi‐cubic and hexagonal sites having a total electron spinS= 1, which were found before in n‐type 4H‐SiC. Based on Q‐band FS ESE and X‐band pulsed ENDOR measurements the values of the hf interaction for Nc1, Nc2were reassigned with respect to the assignment accepted hitherto.The super hyperfine (shf) interaction with neighboring atoms of nitrogen on the quasi‐cubic lattice sites was studied by FS ESE and pulsed ENDOR. The new shf lines due to29Si and13C atoms with large shf interaction which were not observed previously were found in nitrogen FS ESE and ENDOR spectra. One of them, with the largest shf splitting (23.12 MHz), were attributed to the29Si atoms along thec‐axis in nearest‐neighbor positions of nitrogen on the quasi‐cubic sites which unambiguously confirms that nitrogen substitute carbon lattice site in 6H‐SiC. (© 2009 WILEY‐VCH Verlag GmbH &amp; Co. KGaA, Weinheim)

Electron-nuclear double resonance study of [formula omitted] ions in [formula omitted] crystals

Electron paramagnetic resonance (EPR) and electron-nuclear double resonance (ENDOR) are used to identify and characterize V 4+ ions in a bulk single crystal of vanadium dioxide (V O 2). These S = 1 / 2 defects are observed in the as-grown crystal because an adjacent nonmagnetic M 4+ impurity, e.g., a Si 4+ ion, has destroyed the normal antiferromagnetic coupling associated with the close pairs of V 4+ ions that occur in the low-temperature monoclinic phase of V O 2. EPR spectra taken near 5 K with the magnetic field along the [110] and [001] axes show resolved hyperfine patterns due to one 51V nucleus. ENDOR spectra taken at 5 K with the magnetic field parallel to the [001] axis have large nuclear electric quadrupole splittings as a result of a significant electric field gradient at the 51V nucleus. Spin-Hamiltonian parameters describing the electron Zeeman, hyperfine, and nuclear electric quadrupole interactions are reported.

Structure and Properties of 9′-cis Neoxanthin Carotenoid Radicals by Electron Paramagnetic Resonance Measurements and Density Functional Theory Calculations: Present in LHC II?

The radical intermediates formed upon catalytic or photooxidation of the carotenoid 9'-cis neoxanthin inside MCM-41 molecular sieves were detected by pulsed Mims and Davies electron nuclear double resonance (ENDOR) spectroscopies and characterized by density functional theory (DFT) calculations. Mims ENDOR spectra (20 K) were simulated using the hyperfine coupling constants predicted by DFT, which showed that a mixture of carotenoid radical cations (Car(+)) and neutral radicals (#Car) is formed. The DFT relative energies of the neutral radicals formed by proton loss from the C5, C5', C9, C9', C13, and C13'-methyl groups of Car(+) showed that #Car(9') is energetically most favorable, while #Car(9), #Car(13), #Car(13'), #Car(5'), and #Car(5) are less favorable for formation by 2.6, 5.0, 5.1, 22.5, and 25.6 kcal/mol. No evidence for formation of #Car(5') and #Car(5) was observed in the EPR spectra, consistent with DFT calculations. The epoxy group at the prime end and the allene bond at the unprime end prevent protons loss at the C5 and C5'-methyl groups by reducing the conjugation so crucial for the neutral radical stability. Previous CV measurements for allene-substituted carotenoids show that once the radical cations are formed, proton loss is rapid. These examined properties and the known crystal structure of the light harvesting complex II (LHC II) suggest the absence of the neutral radicals of 9'-cis neoxanthin available for quenching the excited states of Chl, consistent with its observed nonquenching properties.

Electron Paramagnetic Resonance and Electron Nuclear Double Resonance Investigation of the Diradical Bis(α-iminopyridinato)zinc Complex

The neutral complex Zn(L*)(2) and its monocationic analogue [ZnL(L*).THF](1+) have been previously reported to contain two and one monoanionic alpha-iminopyridinate((1-)) pi radical ligands, respectively [Lu, C.C.; Bill, E.; Weyhermüller, T.; Bothe, E.; Wieghardt, K. J. Am. Chem. Soc. 2008, 130, 3181-3197] (L represents the neutral alpha-iminopyridine form and THF is tetrahydrofuran). The electronic structures of these complexes have now been studied by Electron Paramagnetic Resonance (EPR) and Electron Nuclear Double Resonance (ENDOR) spectroscopy in conjunction with density-functional theory (DFT) calculations. The ENDOR spectra of the triplet Zn(L*)(2) are characteristic of a localized diradical: these two ligand radicals exhibit dipolar exchange interactions, but no superexchange mechanism is operative, which would be consistent with its nearly tetrahedral coordination geometry. The monocationic species [ZnL(L*).THF](1+) (S = 1/2) has also been investigated by pulse EPR spectroscopy using large interpulse separations. It is shown that no radical hopping takes places on the time scale of the EPR experiment. The results obtained here, in particular the lack of asymmetry in the charge distribution between the two ligands in the triplet state, may be relevant for a better understanding of the electronic structure of naturally occurring diradicals and triplet states.

Electronic structure of the tyrosine D radical and the water-splitting complex from pulsed ENDOR spectroscopy on photosystem II single crystals

Pulsed electron nuclear double resonance (ENDOR) spectroscopy at Q- and W-band frequencies was applied to single crystals of photosystem II from Th. elongatus. W-Band (1)H-ENDOR on the dark-stable radical state Y of the redox-active tyrosine residue Y(D) yields a complete mapping of the electronic structure of this amino acid radical in terms of an assignment of all hyperfine coupling tensors of the protons covalently bound to the side chain. This study can serve as a model case for the potential of high-field/high-frequency ENDOR on protein single crystals for obtaining highly resolved electronic structure information. Q-band (55)Mn-ENDOR was applied to the S(2) oxidation state of the water-splitting complex in photosystem II single crystals. Irrespective of the difficulties arising from the extremely broad electron paramagnetic resonance (EPR) spectroscopy ( approximately 200 mT) and ENDOR ( approximately 100 MHz) spectra a tentative assignment of the Mn ion in the formal oxidation state III to a Mn position in the structural model of PSII is possible on the basis of the ENDOR data.

Population transfer for signal enhancement in pulsed EPR experiments on half integer high spin systems

High resolution pulse EPR techniques applied to half integer high spin systems, such as Mn(2+) (S = 5/2), usually focus only on the central |-1/2--> |1/2 transition. The reason is that at high fields, where the zero field splitting is considerably smaller than the Zeeman interaction, the spectrum of this transition is intense and narrow. However, because the experiments are carried out at low temperatures, the low lying levels are heavily populated and the signal of the central transition is nevertheless diminished. This, in turn affects the sensitivity of the pulse EPR technique applied. A transfer of populations from the lower lying levels, which for Mn(2+) are the |-3/2 and |-5/2 levels, to the |-1/2 level will therefore increase the sensitivity. Here we describe such an experiment, where a rapid magnetic field sweep over the |-3/2--> |-1/2 sub-spectrum is carried out, concomitantly with a low power microwave (mw) irradiation, which results in population inversion. After this sweep any pulsed EPR sequence can be applied to the central transition that now has a population difference that deviates from the equilibrium value. The feasibility of the experiment is demonstrated at W-band (95 GHz) on Mn(2+) doped in MgO for echo-detected EPR measurements and the dependence of the signal enhancement on the rate and range of the magnetic field sweep and on the mw power is described. The results are then accounted for theoretically by considering a simple fictitious spin 1/2 system. In addition, preliminary enhanced (55)Mn pulse ENDOR electron nuclear double resonance (ENDOR) spectra are presented.