High-frequency EPR studies on cofactor radicals in photosystem I

Electron paramagnetic resonance (EPR) spectroscopy at W-band (94 GHz) is used to resolve theg-tensors of the radical ions of the primary chlorophyll donor P ₇₀₀ ^+? and the quinone acceptor A ₁ ^?? in photosystem I. The obtainedg-tensor principal values are compared with those of the isolated pigment radicals in organic solvents and the shifts are related to an impact of the protein environment. P ₇₀₀ ^+? has been investigated in photosystem I single crystals at 94 GHz. W-band EPR applied to the photoinduced radical pair P ₇₀₀ ^+? A ₁ ^?? is used to correctly assign theg-tensor axes of P ₇₀₀ ^+? to the molecular structure of the primary donor. Density functional theory calculations on a model of A ₁ ^?? in its binding pocket derived from the recent crystal structure of photosystem I were utilized to correlate experimental magnetic resonance parameters with structural elements of the protein.     

Theg-tensor anisotropy of the triplet state of the primary electron donor in the photosynthetic bacteriumRhodobacter sphaeroides by high-field (95 GHz) EPR

The anisotropy of theg-tensor of the light-induced triplet state of the primary electron donor (D) of the photosynthetic bacteriumRhodobacter sphaeroides is determined by electron-spin-echo-detected electron paramagnetic resonance at 95 GHz. Measurements on frozen solutions of quinone-depleted reaction centers yield g-values along the principal directions of the zero-field splitting tensor (Norris J.R., Budil D.E., Gast P., Chang C.H., ElKabbani O., Schiffer M.: Proc. Natl. Acad. Sci. USA86, 4335–4339, 1989).g-Values determined are: 2.00308, 2.00238, and 2.00138. The deviation from axial symmetry and thez-component of the g-tensor are smaller than observed in the cation radical of D.     

High-field/high-frequency EPR/ENDOR — A powerful new tool in photosynthesis research

In photosynthesis research the elucidation of the spatial and electronic structures of the electron donor and acceptor ion radicals is very important for an understanding of the light-induced electron transfer process. Recent 3 mm (W-band, 95 GHz) high-field EPR and ENDOR studies on the primary donor cation radicals P^+. (bacteriochlorophyll dimer), the acceptor anion radicals Q^+. (quinones), and the charge-separated radical pair (P^+.Q^?.) in photosynthetic bacteria and biomimetic model systems are presented. From single crystals of, for instance,Rb. sphaeroides R-26 reaction centers both the hyperfine tensors of various protons and theg-tensor of P₈₆₅ ^+. have been determined and compared with calculated tensor values based on recent X-ray structure data. The results consistently reveal a breaking of the local C₂ symmetry of the electronic structure at the primary donor side of the reaction center. This is of particular interest since it might be relevant for the vectorial electron transfer along the protein complex. Among the quinone radical anions studied are frozen solutions of the electron acceptors of bacterial and plant reaction centers (ubiquinone and plastoquinone, respectively). The increased electron Zeeman interaction in high-field EPR leads to almost completely resolvedg-tensor components even in disordered samples. Theg-tensors and component linewidths are sensitive probes for specific anisotropic interactions with the environment. In the case of the transient correlated coupled radical pair P₈₆₅ ^+.-Q_A ^?. ofRb. sphaeroides (Fe replaced by Zn) the spin-polarized high-field EPR spectra allow an unambiguous determination of the relative orientation of theg-tensors of the donor and acceptor parts. Thereby high-precision structure information is obtained on the electron transfer pigments after light-induced charge separation.     

Transient EPR at 240 GHz of the excited triplet state of free-base tetra-phenyl porphyrin

The electron paramagnetic resonance (EPR) spectrum measured at 240 GHz of the lowest excited triplet state of the free-base tetra-phenyl porphyrin (H₂-TPP) molecule is presented. The high frequency that is employed allows for the full resolution of theg-anisotropy (2.00354(2), 2.00272(15), and 2.00194(5) forx, y, andz zero-field axes respectively), and the obtained values are compared to those of other porphyrin-based systems. These results are one of the first addressing theg-anisotropy in the photoexcited state of porphyrins. The usefulness of high-field transient EPR in elucidating not only the principal values of theg-tensor but also the orientation of theg-tensor principal axes with respect to those of the zero-field splitting tensor in porphyrin disordered systems is also discussed.     

Influence of hydrogen bonds on the electronicg-tensor and13C-hyperfine tensors of13C-labeled ubiquinones — EPR and ENDOR study

Selectively^l3C-labeled ubiquinone anion radicals in protic and aprotic solvents are investigated by EPR and ENDOR spectroscopy, yielding information about the effect of hydrogen bonds on the electronicg-tensor and the carbonyl carbon¹³C-hf tensors. Formation of the hydrogen bonds alter theg-tensor significantly to lower values and increases theA _zz , component of the^l3C-hf tensor. Both effects can be explained by electrostatic interactions between the positively charged hydrogen and the electrons at the carbonyl oxygen leading to a redistribution of charge and π-spin density. Two different hydrogen bonds were obtained for UQ ₀ ^? which are in agreement with the results of DFT (density functional theory) calculations.     

Spin-label HF-EPR of lipid ordering in cholesterol-containing membranes

Conventional oxazolidine spin-labelled lipids have the axial¹⁴N-hyperfine tensorz-axis directed along the long axis of the lipid chain. Investigation of lateral ordering of the lipids in membranes requires measurement of thex-y Zeeman anisotropy of the nonaxialg-tensor at high fields. Both the lateral and transverse ordering of the lipid chains in membranes of dimyristoyl phosphatidylcholine containing 40 mol% cholesterol in the liquid-ordered phase have been studied with 94 GHz electron paramagnetic resonance spectroscopy. This has been done by using probe amounts of phosphatidylcholine systematically spin-labelled at positionsn along the length of thesn- 2 chain [n-PCSL, 1-acyl-2-(n-(4,4-dimethyloxazolidine-N-oxyl) stearoyl)-sn-glycero-3-phosphocholine]. Nonaxial (g_xx?g_yy) anisotropy of the spin-labelled lipid chains is detected over a wide range of temperature throughout the liquid-ordered phase. The transverse profile of lateral ordering with position,n, of chain labelling follows the profile of the rigid steroid nucleus of cholesterol. It becomes progressively averaged towards the terminal methyl group of thesn- 2 chain, in the region of the flexible hydrocarbon chain of cholesterol. The nonaxial lipid ordering may be related to lipid domain formation in membranes containing cholesterol and saturated-chain lipids.     

Optical detected ESR in the4 S 3/2 excited state of Er:YCl3

The electron spin resonance in the⁴ S _3/2 excited state of Er³⁺ in yttrium trichloride was studied by optical detection techniques. From angular dependence of the resonance field the principle value of theg-tensor in direction of the twofold crystal axis was deduced to beg∥=3.350±0.004 and the perpendicular valueg⊥ in the crystallographica-b-plane was extrapolated to beg⊥=2.857±0.004. The lifetime of the excited state is found to be temperature independent with τ _r =(1.62±0.02)·10^?3 sec and the spin lattice relaxation timeT ₁ was determined in the temperature region 1.5 to 2.1 °K by observing the recovery of the fluorescent light signal after a microwave saturation pulse was switched off.T ₁ is found to follow a direct process with \(T_1^{ - 1} = k \cdot cth\left( {\frac{{\rlap{--} h\omega }}{{2kT}}} \right)\) .     

Elektronenspin-Resonanz von Stickstoffzentren in Alkalihalogenid-Kristallen

Paramagnetic nitrogen centers are produced in nitrate doped single crystals of KCl, KBr, KJ, and NaCl by introduction of F-centers. The electron spin resonance is studied at low temperature. The hyperfine structure of the lines is caused by interaction with two N¹⁴ nuclei. The angle dependence of the resonance spectra is measured by rotating the crystals about several crystallographic axes. The results can be fitted to a spin-Hamiltonian with orthorhombic symmetry. The components of theg-tensor and theA-tensor are determined. The centers are believed to be N ₂ ^? -ions in negative ion vacancies.     

Corrigendum

Theoretical calculations of g-tensor components for the spin–orbit quartet, which arises as the ground state in three-coordinate d⁹ complexes and low-spin d⁷ complexes of D_3h symmetry, have been made on the assumption that the spin–orbit interaction is commensurable with the electron-vibrational interaction. The calculations were carried out within the framework of crystal field theory using representations of the hole formalism. The analytical expressions for g-tensor components were obtained limited to first-order terms. It was shown that the account of the electron–vibrational interaction in the excited quartet only provides three-axial anisotropy for the g-tensor. It was shown that the g-tensor rotates in the plane of the three-coordinate structure with consensual motion of the atoms. The resulting expressions for the g-factor components are in good agreement with experimental data. Being universal for a wide range of contributions of the vibronic and spin–orbit interactions, these expressions essentially fill the gap in studying structures of coordination compounds.     

ESR study of rare-earth ions with the effective spin of 1/2 in LiNbO3 crystals

Rare-earth ions of Nd³⁺ and Er³⁺ in nearly stoichiometric and MgO-doped LiNbO₃ crystals, respectively, have been investigated by employing an X-band electron spin resonance (ESR) spectrometer. The grown crystal was heated in Li-rich powder at 1100°C in order to make it nearly stoichiometric by the vapor transport equilibrium technique. Due to the fact that the ESR linewidth is much narrower in the stoichiometric crystal than in the congruent LiNbO₃, we were able to determine the hyperfine constants of¹⁴³Nd and¹⁴⁵Nd at 4 K. By codoping MgO into LiNbO₃, a new Er³⁺ center has been observed with a differentg-tensor. We propose that the new Er³⁺ center in Mg-doped LiNbO₃ occupies the niobium site due to the local excessive Mg²⁺ ion at the lithium site, whereas Nd³⁺ and Er³⁺ in congruent crystals reside at the lithium site. The proposal is consistent with theg-value anisotropy.     

Paramagnetic centres in tungstate powders

The EPR-powder spectrum ofV _k -centres in CaWO₄ is simulated using the experimentally obtained principal values of theg-tensor. The influence of the finite linewidth ΔH on the resulting EPR-pattern is taken into account. With these results the existence ofV _k -centres in x-rayed microcrystalline tungstate powder is proved. The resonance of the² S _1/2-state of Pb³⁺-ions substitutionally incorporated on Ca²⁺-sites in CaWO₄ is measured in the powder spectrum. The optically and thermally stimulated valence state changes of the centres are studied and are shown to be correlated with the thermoluminescence glow curve.     

On Electron Spin Polarization Created in the Excited Triplet State of Accessory Chlorophyll via Photoinduced Charge-Recombination of the Photosystem II Reaction Center

We present a theoretical approach to investigate the electron spin polarization (ESP) of the excited triplet state that has been detected using the time-resolved electron paramagnetic resonance (TREPR) method in the photosystem II center of the plants. We show, using the stochastic Liouville equation, that the ESP pattern created in the accessory chlorophyll (Chl_accD1) which reside near the P_D1 chlorophyll of the active branch is explained by one-step, concerted double electron transfer model, initiating from the singlet–triplet conversion of the light-induced charge-separated state composed of P_D1 radical cation and pheophytin radical anion. We also considered the sequential ESP transfer model via the triplet charge-recombination (CR) and the triplet–triplet energy transfer processes. It has been clearly shown that the ESP created in the ³Chl_accD1* is dependent on the rate constant (k _TT) of the triplet–triplet energy transfer from the intermediate triplet state created by the CR. Also we show that the relative orientation of the principal axes of the spin dipolar interaction in the intermediate triplet state (³P_D1*, as an example) may play a role in the ESP pattern, when the k _TT is smaller than the angular frequency of the Zeeman energy. We have theoretically shown that the TREPR measurement of the ESP is very powerful to investigate the primary chemical process and to characterize the intermediate as a signature of the stepwise ESP transfer.     

Temperature-dependent electron spin polarization of the triplet state of the primary donor in photosynthetic reaction centers ofRhodopseudomonas viridis: A time-resolved EPR study of single crystals and solid solution

The electron spin polarization (ESP) of triplet of the primary donor (³P) ofRhodopseudomonas viridis reaction centers (RCs) is anomalous at temperatures above 25 K, i.e. the steady-state ESP changes from AEEAAE to AEAEAE. Fast, time-resolved EPR measurements in solid solution and single crystals of RCs show that this phenomenon results most probably from fast anisotropic spin-lattice relaxation in the radical pair triplet state (k _r≈ 5·10⁹s^?1 at 25 K).     

Towards an identification of chemically different flavin radicals by means of theirg-tensor

Theg-tensors of two chemically different flavin mononucleotide (FMN) radicals, one of which is covalently bound via N(5) of its 7,8-dimethyl isoalloxazine moiety, and the other one non-covalently bound to mutant LOV domains of the blue-light receptor phototropin, LOV1 C57M and LOV2 C450A, respectively, have been determined by very high microwave frequency and high magnetic field electron paramagnetic resonance (EPR) performed at 360 GHz and 12.8 T. Due to the high spectral resolution of the frozen-solution continuous-wave EPR spectra, the anisotropy of theg-tensors could be fully resolved. By least-squares fittings of spectral simulations to expermental data, the principal values ofg have been established:g _X=2.00554(5),g _Y=2.00391(5), andg _Z=2.00247(7) for the N(5)-alkyl-chain-linked FMN radical in LOV1 C57M-675, andg _X=2.00427(5),g _Y=2.00360(5), andg _Z=2.00220(7) for the noncovalently bound FMN radical in LOV2 C450A-605. By a comparison of these values to the ones from the flavin adenine dinucleotide radicals in two photolyases, the radical in LOV2 C450A-605 could be clearly identified as a neutral FMN radical, FMNH. In contrast, LOV1 C57M-675 exhibits significantly shifted principal components ofg, the differences being caused by spin-orbit coupling of the nearby sulfur from the reactive methionine residue, and the modified chemical structure due to the covalent attachment at N(5) of the radical to the apoprotein. The results clearly show the potential of using theg-tensor as probe of the global electronic and chemical structure of protein-bound flavin radicals.     

Shifts of g Values in the Excited Triplet States of Metal Complexes Studied by Time-Resolved W-band EPR

A highly time-resolved high-frequency/high-field W-band electron paramagnetic resonance (EPR) (ν ~ 94 GHz) is a powerful technique to determine small g anisotropies of transient paramagnetic species. We applied this method to studies of the lowest excited triplet (T₁)³ ππ* states in metal complexes such as a platinum (Pt) diimine complex and metal (Zn and Mg) porphines in rigid glasses. From the analyses of time-resolved EPR spectra, g anisotropies were obtained as g _z  = 2.0048, g _x  = g _y  = 2.0035 for Pt(b-iq)(CN)₂ (b-iq = 3,3′bi-isoquinoline) and g _z  = 1.9968, g _x  = g _y  = 2.0022 for zinc tetraphenylporphine (ZnTPP). No measurable anisotropies were found for magnesium (Mg) TPP. The g values of the Pt complex are larger than g _e (=2.0023, g value of free electron) and that g _z of ZnTPP is smaller than g _e. These results were interpreted in terms of the nature of the perturbed states: the higher triplet ππ′* state mixes with T₁(ππ*) via spin–orbit coupling in ZnTPP. In contrast, the higher triplet dπ* state is involved in this coupling for the Pt complex. Thus, the nature of the perturbed state can be distinguished from the anisotropic g values of the T₁(ππ*) state.     

Bestimmung desg J -Faktors im 4s4p 3 P 1-Term des CaI-Spektrums mit optischer Doppelresonanz

The³ P ₁ state of Ca (τ≈0,5 msec) was populated by resonance-absorption of the intercombination line (λ=6572,8 Å). The radio-frequency-transitions between the Zeeman-sublevels of the excited state were observed by means of optical double-resonance. Because of the oscillator-strength of the intercombination line being extremely small, a second excitation method, namely with electron impact, was tried. Both methods yielded equivalent results. In order to determine theg _J -value of the³ P ₁-state the static magnetic field was calibrated by Zeeman-transitions in the ground state of Na, the transitions being detected by optical pumping,g _J was found to be 1,50105(7) in agreement with the theoretical value 1,50108.     

Selective non-radiative transitions at excited states of the self-trapped exciton in alkali halides

Generation of F-H pairs and σ-luminescence induced by excitation of the self-trapped excitons with polarized light causing the 1s→ 2p transitions has been measured. The results were analyzed based on the assumption that the non-radiative transitions that follow photoexcitation depend only on the state reached by the excitation, irrespective of the photon energy of the excitation. The absence of the dependence on exciting photon energy of the effective yield of removal from the triplet manifold after excitation to each substate obtained from the analysis proves that the above assumption is valid. The relative non-radiative transition probabilities between the 2p substates and from the 2p-substates to the lowest triplet state, the F-H pair, the σ-luminescent state and the ground state were obtained. It is shown that the de-excitation channels from each substate are substantially different from each other. The following transitions are found to have high probabilities: from the B_1g state to the F-H pairs in KCl and KBr, from theA_g state to the σ-luminescent state and the lowest triplet state in KBr and to the lowest triplet state in KCl and from the B_2g state to the B_1g state in KCl and KBr, where A_g, B_1g and B_2g denote the states with the electron excited to the σ_u orbital, the π_u orbital lying in the (100) plane in which the (halogen)₂^?-molecular ion is situated and the other π_u orbital, respectively. The mechanisms of these non-radiative transitions were discussed.     

Levelcrossing experiments in the first excited1 P 1 states of the alkaline earths

The hyperfine structure of the lowest¹P₁ state of²⁵Mg,⁴³Ca,⁸⁷Sr,¹³⁵Ba and¹³⁷Ba have been measured by the level-crossing and anticrossing technique. The magnetic dipole and electric quadrupole coupling constants determined by these measurements are²⁵Mg(3s3p¹P₁):A=? 7.7(5) MHz; 16 MHz>B>0 MHz,⁴³Ca(4s4p¹P₁):A=? 15.3(4) MHz; ¦B¦<12 MHz,⁸⁷Sr (5s5p¹P₁:A=? 3.4(4) MHz;B=39(4) MHz,¹³⁵Ba(6s6p¹P₁):A=? 97.5(1.0) MHz;B=31(9)MHz,¹³⁷Ba(6s6p¹P₁):A=?109.2(1.2) MHz;B=51(12)MHz. The results have been compared with the predictions of the Breit-Wills theory of the two-electron hyperfine structure using the experimental data on the³P states. Large discrepancies have been observed which are due to different radial wave functions of thes andp electron in the triplet and singlet system. This effect has been taken into account by fitting the data with the aid of two additional parameters. That this procedure is justified is shown by an analysis of the fine structure splitting, the life times, and the isotopic shifts in thesp configurations of group II elements.     

Zur Rumpfpolarisation des Rb-Atoms: Hyperfeinstruktur des 72 P 1/2-Terms von Rb85

The hyperfine structure splitting of the 7² P _1/2 state of Rb⁸⁵ has been investigated with optical double resonance spectroscopy. The results are:v _F=3?F=2=52.95(6) Mc/s,A _J=1/2=17.65(2) Mc/s,g _J (7² P _1/2, Rb)=0.6668(1). From the hfs interaction constants of the 7² P multiplett a 12% core polarization contribution to the magnetic hfs of the 7² P _3/2 state can be deduced. Comparison is made between the values of 〈r ^?3〉 calculated from either the magnetic hfs or the² P fine structure separation.     

Optical-Optical Double Resonance Spectroscopy of the 1g(P1)-AΠ (1u)-XΣg Transition of I2

Optical-optical double resonance spectroscopy was used to study the 1_g(³P₁) ion-pair state of I₂ correlating to I⁻(¹S)+I⁺³P₁) at the dissociation limit. We gained access to the 1_g(³P₁) state though the A³Π (1_u) state in the (1+1) photon-excitation scheme. The pump laser excited the A³Π (1_u)-X¹Σ_g⁺ transition at a fixed frequency for state selection. The probe laser was scanned to detect the 1_g(³P₁)-A³Π (1_u) resonance by monitoring the ultraviolet emission from the 1_g(³P₁) state at 278 nm. The 1_g(³P₁) state was observed in a vibrational progression consisting of P and R doublets. An energy level analysis was carried out for the 1_g(³P₁) state in the 0≤ v ≤ 14 and 12≤J≤135 range, which led to a set of molecular parameters including the Ω-doubling constant. The Ω-doubling of the 1_g(³P₁) state was discussed by the pure precession model and interpreted to occur through the heterogeneous coupling with the 0_g⁻(³P₁) state correlating to the same ionic asymptote.