Water Activity Regulates the Q(A)(-) to Q(B) electron transfer in photosynthetic reaction centers from Rhodobacter sphaeroides

We report on the effects of water activity and surrounding viscosity on electron transfer reactions taking place within a membrane protein: the reaction center (RC) from the photosynthetic bacterium Rhodobacter sphaeroides. We measured the kinetics of charge recombination between the primary photoxidized donor (P(+)) and the reduced quinone acceptors. Water activity (aW) and viscosity (eta) have been tuned by changing the concentration of cosolutes (trehalose, sucrose, glucose, and glycerol) and the temperature. The temperature dependence of the rate of charge recombination between the reduced primary quinone, Q(A)(-), and P(+) was found to be unaffected by the presence of cosolutes. At variance, the kinetics of charge recombination between the reduced secondary quinone (Q(B)(-)) and P(+) was found to be severely influenced by the presence of cosolutes and by the temperature. Results collected over a wide eta-range (2 orders of magnitude) demonstrate that the rate of P(+)Q(B)(-) recombination is uncorrelated to the solution viscosity. The kinetics of P(+)Q(B)(-) recombination depends on the P(+)Q(A)(-)Q(B) <--> P(+)Q(A)Q(B)(-) equilibrium constant. Accordingly, the dependence of the interquinone electron transfer equilibrium constant on T and aW has been explained by assuming that the transfer of one electron from Q(A)(-) to Q(B) is associated with the release of about three water molecules by the RC. This implies that the interquinone electron transfer involves at least two RC substates differing in the stoichiometry of interacting water molecules.     

Simulation of ultrafast non-exponential fluorescence decay induced by electron transfer in FMN binding protein

The ultrafast non-exponential fluorescence decay of FMN binding protein (FBP) was analyzed with three electron transfer (ET) theories, Marcus theory, Bixon and Jortner theory and Kakitani and Mataga theory. Center to center distances between electron acceptor, the excited isoalloxazine, and donors, Trp-32, Tyr-35 and Trp-106, in FBP were determined by molecular dynamic simulation. Electron transfer parameters containing in these theories were determined so as to fit the calculated decay with the observed decay, according to a non-linear least squares method. Introduction of electrostatic energies between isoalloxazine anion and other ionic groups and between the donor cations and other ionic groups in the protein into any ET theories improved the fitting. The non-exponential behavior in the fluorescence decay is considered to be ascribed to a fluctuation of the protein structure with long period.     

Evidence for concerted electron proton transfer in charge recombination between FADH- and 306Trp• in Escherichia coli photolyase

Proton-coupled electron-transfer (PCET) is a mechanism of great importance in protein electron transfer and enzyme catalysis, and the involvement of aromatic amino acids in this process is of much interest. The DNA repair enzyme photolyase provides a natural system that allows for the study of PCET using a neutral radical tryptophan (Trp(?)). In Escherichia coli photolyase, photoreduction of the flavin adenine dinucleotide (FAD) cofactor in its neutral radical semiquinone form (FADH(?)) results in the formation of FADH(-) and (306)Trp(?). Charge recombination between these two intermediates requires the uptake of a proton by (306)Trp(?). The rate constant of charge recombination has been measured as a function of temperature in the pH range from 5.5 to 10.0, and the data are analyzed with both classical Marcus and semi-classical Hopfield electron transfer theory. The reorganization energy associated with the charge recombination process shows a pH dependence ranging from 2.3 eV at pH ≤ 7 and 1.2 eV at pH(D) 10.0. These findings indicate that at least two mechanisms are involved in the charge recombination reaction. Global analysis of the data supports the hypothesis that PCET during charge recombination can follow two different mechanisms with an apparent switch around pH 6.5. At lower pH, concerted electron proton transfer (CEPT) is the favorable mechanism with a reorganization energy of 2.1-2.3 eV. At higher pH, a sequential mechanism becomes dominant with rate-limiting electron-transfer followed by proton uptake which has a reorganization energy of 1.0-1.3 eV. The observed 'inverse' deuterium isotope effect at pH < 8 can be explained by a solvent isotope effect that affects the free energy change of the reaction and masks the normal, mass-related kinetic isotope effect that is expected for a CEPT mechanism. To the best of our knowledge, this is the first time that a switch in PCET mechanism has been observed in a protein.     

Light emission originating from photosystem II radical pair recombination is sensitive to zeaxanthin related non-photochemical quenching (NPQ)

We have used chlorophyll fluorescence, delayed luminescence and thermoluminescence measurements to study the influence of an artificial DeltapH in the presence or absence of zeaxanthin on photosystem II reactions. Energization of the pea thylakoid membranes induced non-photochemical fluorescence quenching and an increase in the overall luminescence emission of PSII during delayed luminescence and thermoluminescence measurements. This DeltapH-induced overall luminescence increase was caused by a strongly enhanced delayed luminescence in the seconds range before sample heating. In the subsequent thermoluminescence measurements the intensity of the B-band decreased after one and increased after two or more single turnover flashes. We propose that strong membrane energization shifted the redox potential of photosystem II radical pairs to more negative values causing the high delayed luminescence. The zeaxanthin-dependent non-photochemical fluorescence quenching component, however, did not alter thermoluminescence B-bands but decreased the delayed luminescence intensity by 30%. To our knowledge this is the first report that the radiative radical pair recombination, exhibited as delayed luminescence but not thermoluminescence emission, is sensitive to the antenna located zeaxanthin related non-photochemical fluorescence quenching. Our data can be interpreted within the frame of the exciton/radical pair equilibrium model that describes photosystem II as a shallow trap and incorporates the transfer of energy from the re-excitated reaction centre to the antenna of photosystem II.     

Afterglow thermoluminescence measured in isolated chloroplasts

The thermoluminescence afterglow (AG) measured in plant leaves originates from the S(2)/S(3)Q(B)(-) charge pair recombination in photosystem II (PSII) initiated by reverse electron flow from stromal reductants to PQ and then to the Q(B) site in PSII centers that are in the S(2)/S(3)Q(B) state. In this study, we show that this luminescence, absent in isolated thylakoid membranes, can be measured in intact chloroplasts that retain their stromal content including the electron acceptor pool (oxidized ferredoxin/NADP(+)) of photosystem I. The properties of the chloroplasts AG emission is similar to the AG in leaves in terms of temperature maximum, period-four modulation, far-red light stimulation, and antimycin A inhibition.     

Marcus电子转移理论的科学性探讨

利用科学原理对Marcus电子转移理论的科学性进行了考察, 结果表明Marcus电子转移理论违背了能量守恒定律.     

PHOTOCHEMICAL AND SPECTRAL PROPERTIES OF A PARTICULATE MODEL SYSTEM OF CHLOROPHYLL WITH AMPHIPHILES PREPARED FROM HISTAMINE

Abstract— In the photosynthesis model system described, chlorophyll a at an interface photosensitizes the transfer of hydrogen equivalents from a hydrocarbon phase to an aqueous phase. The hydrocarbon phase, to which chlorophyll is adsorbed, consists of polyethylene particles swollen with tetradecane. The particles are also charged positive by co-adsorption of dodecylpyridinium iodide. Furthermore, chlorophyll is ligated with the imidazole function of one of several amphiphiles derived from histamine, which may or may not contain a reducible nitroaromatic group that can serve as primary electron acceptor from photoexcited chlorophyll. The fluorescence quantum yield of chlorophyll on these particles is diminished by self-association of the pigment and by reaction with an oxidizing amphiphile; in the latter case, the quantum yield is correlated with the one-electron redox potential of the amphiphile. Fluorescence-lifetime analysis reveals that most excited singlet states of chlorophyll are quenched rather quickly, and that most of the fluorescence comes from a small fraction of chlorophyll with long lifetime. All preparations sensitize the photoreduction of 5,5′-dithiobis(2-nitrobenzoate) (DTNB) to the water-soluble thiolate by hydrazobenzene. When the amphiphile that ligates chlorophyll is not oxidizing, the quantum yield of photoreduction is unrelated to the fluorescence yield of the particles, but may be related to the degree of self-association of chlorophyll. When the amphiphile that ligates chlorophyll is oxidizing, the kinetics of photoreduction of DTNB require that the electron passes through the primary oxidant to DTNB. Quantum yields for photosensitized reducton of oxidizing amphiphiles in the absence of DTNB have a reversed correlation with redox potential, which can be rationalized in terms of the Marcus theory of electron transfer. All data are most consistently accounted for if the primary photoproduct is an ion pair of chlorophyll and primary oxidant when the latter is available, and a chlorophyll radical ion pair when it is not, both formed by electron transfer from the singlet excited state of chlorophyll.     

Measurements and modeling of recombination from nanoparticle TiO2 electrodes

Electron-transfer reactions from nanoparticle TiO(2) films to outer-sphere redox shuttles were investigated. Steady-state dark current density versus applied potential and open circuit voltage decay measurements were employed to determine the rates of recombination to cobalt(III) tris(4,4'-dimethyl-2,2'-bipyridyl), [Co(Me(2)bpy)(3)](3+), and ruthenium(III) bis(2,2'-bipyridyl)-bis(N-methylimidozole), [Ru(bpy)(2)(MeIm)(2)](3+). A striking difference in the magnitude as well as the shape of the electron lifetimes for TiO(2) electrodes in contact with these two redox shuttles is observed. A model based on Marcus theory is developed to describe recombination, including contributions from conduction band electrons and surface states. Excellent agreement was found between the modeled and measured lifetimes. The model allows for identification of each contributing component of electron transfer to the measured lifetimes. Comparison of the different components of the modeled lifetimes to the measured lifetimes provides clear evidence for recombination mediated through surface states.     

Photoinduced electron transfer and geminate recombination in liquids on short time scales: Experiments and theory

The coupled processes of intermolecular photoinduced forward electron transfer and geminate recombination between the (hole) donor (Rhodamine 3B) and (hole) acceptors (N,N-dimethylaniline) are studied in three molecular liquids: acetonitrile, butyronitrile, and benzonitrile. Two color pump-probe experiments on time scales from approximately 100 fs to hundreds of picoseconds give information about the depletion of the donor excited state due to forward electron transfer and the survival kinetics of the radicals produced by forward electron transfer. The data are analyzed with a model presented previously that includes distance dependent forward and back electron transfer rates, donor and acceptor diffusion, solvent structure, and the hydrodynamic effect in a mean-field theory of through solvent electron transfer. The forward electron transfer is in the normal regime, and the Marcus equation for the distance dependence of the transfer rate is used. The forward electron transfer data for several concentrations in the three solvents are fitted to the theory with a single adjustable parameter, the electronic coupling matrix element Jf at contact. Within experimental error all concentrations in all three solvents are fitted with the same value of Jf. The geminate recombination (back transfer) is in the inverted region, and semiclassical treatment developed by Jortner [J. Chem. Phys. 64, 4860 (1976)] is used to describe the distance dependence of the back electron transfer. The data are fitted with the single adjustable parameter Jb. It is found that the value of Jb decreases as the solvent viscosity increases. Possible explanations are discussed.     

半菁染料作为染料敏化太阳能电池吸光材料的理论研究

采用第一性原理研究了半菁-二氧化钛团簇形成的配合物(hemicyanine-(TiO₂)_n)的光电子转移过程, 这里n分别取5, 9, 15. 配合物基态构型采用密度泛函理论方法进行优化, 而激发态采用含时密度泛函理论进行计算. 采用长程相关校正的密度泛函CAM-B3LYP和ωB97X-D计算的激发能与实验值吻合得很好. 依据广义Mulliken-Hush (GMH)公式, 基于密度泛函理论得到的波函数被用来计算电荷转移积分, 进而可根据Marcus理论计算出电荷分离速率常数(k_CS)和电荷回传速率常数(k_CR). 计算结果表明电子从染料到(TiO₂)_n团簇的传递有多条通道, 这使得k_CS具有更大值, 相反, 只具有单通道的电荷回传降低了k_CR值, 与k_CS相比甚至可以忽略, 这表明在所研究的体系中电荷回传是不利的.     

Electron transfer in porphyrin—quinone cyclophanes studied on the pico- and femto-second time scale

Electron transfer in porphyrin—quinone cyclophanes is investigated by fluorescence and absorption spectroscopy with pico- and femto-second pulses. In nonpolar solvents, the S₁ state of the porphyrin shows a lifetime from 300 ps up to several nanoseconds, depending upon the number of quinones and upon their electron affinity. Comparative measurements in polar solvents demonstrate very fast electron transfer on a time scale between 1 and 10 ps. The results are analyzed with the aid of quantum-chemical calculations which give the energy of the charge transfer states and the relevant coupling strengths. For nonpolar solvents, theory suggests fluctuation-induced charge separation and/or direct radiationless internal conversion from the porphyrin S₁ to the ground state. In polar solution, the molecules exist in a tilted configuration with strong electronic coupling and charge transfer states well below the S₁ level, resulting in fast electron transfer and subsequent charge recombination within 10–40 ps.     

Primary processes in plant photosynthesis: photosystem I reaction center

The photosystem I (PSI) pigment-protein complex of plants converts light energy into a transmembrane charge separation, which ultimately leads to the reduction of carbon dioxide. Recent studies on the dynamics of primary energy transfer, charge separation, and following electron transfer of the reaction center (RC) of the PSI prepared from spinach are reviewed. The main results of femtosecond transient absorption and fluorescence spectroscopies as applied to the P700-enchied PSI RC are summarized. This specially prepared material contains only 12–14 chlorophylls per P700, which is a special pair of chlorophyll a and has a significant role in primary charge separation. The P700-enriched particles are useful to study dynamics of cofactors, since about 100 light-harvesting chlorophylls are associated with wild PSI RC and prevent one from observing the elementary steps of the charge separation. In PSI RC energy and electron transfer were found to be strongly coupled and an ultrafast up-hill energy equilibration and charge separation were observed upon preferential excitation of P700. The secondary electron-transfer dynamics from the reduced primary electron acceptor chlorophyll a to quinone are described. With creating free energy differences (ΔG₀) for the reaction by reconstituting various artificial quinones and quinoids, the rate of electron transfer was measured. Analysis of rates versus ΔG₀ according to the quantum theory of electron transfer gave the reorganization energy, electronic coupling energy and other factors. It was shown that the natural quinones are optimized in the photosynthetic protein complexes. The above results were compared with those of photosynthetic purple bacteria, of which the structure and functions have been studied most.     

Synthesis and photophysics of monodisperse co-oligomers consisting of alternating thiophene and perylene bisimide

A series of monodisperse oligomers consisting of alternating thiophene (T) and perylene bisimide (P), denoted as (TP)(n)T (n = 1, 2, 3, 6), were synthesized and photophysically characterized. The steady-state absorption and fluorescence spectra revealed that the low-energy P-derived band remains almost unchanged upon the increment of the number of the repeat unit n. This can be rationalized as a consequence of nearly orthogonal molecular geometry and highly-localized electron density at LUMO level based on DFT calculation. A drastic reduction of the fluorescence quantum yields (Φ(F)) of (TP)(n)T was observed with the sequence of (TP)(6)T > (TP)(3)T > (TP)(2)T > (TP)(1)T, as compared to the parent perylene bisimide. Further femtosecond transient absorption studies clarified that the quenching mechanism is intramolecular electron transfer, in which the generated P radical anion was spectrally recognized. The rate of charge separation was found to be on the order of 10(11) s(-1), suggesting an efficient electron transfer reaction between the thiophene and perylene units. Interestingly, the charge separation rate constant increased more than three times upon the increment of n, whereas the charge-recombination rate constant remained almost unchanged at (1.58-2.21) × 10(9) s(-1). Analysis of the kinetic and thermodynamic data using the Marcus approach showed that the enhanced electronic coupling is the origin of the acceleration of electron-transfer reaction in the D-A copolymers.     

New perspective of electron transfer chemistry

A new perspective of electron transfer chemistry is described for fine control of electron transfer reactions including back electron transfer in the charge separated state of artificial photosynthetic compounds and its synthetic application. Fundamental electron transfer properties of suitable components of efficient electron transfer systems are described in light of the Marcus theory of electron transfer, in particular focusing on the Marcus inverted region, and they are applied to design multi-step electron transfer systems which can well mimic the function of a photosynthetic reaction center. Both intermolecular and intramolecular electron transfer processes are finely controlled by complexation of radical anions, produced in the electron transfer, with metal ions which act as Lewis acids. Quantitative measures to determine the Lewis acidity of a variety of metal ions are given in relation to the promoting effects of metal ions on the electron transfer reactions. The mechanistic viability of metal ion catalysis in electron transfer reactions is demonstrated by a variety of examples of chemical transformations involving metal ion-promoted electron transfer processes as the rate-determining steps, which are made possible by complexation of radical anions with metal ions.     

A schematic model for energy and charge transfer in the chlorophyll complex

A theory for simultaneous charge and energy transfer in the carotenoid-chlorophyll-a complex is presented here and discussed. The observed charge transfer process in these chloroplast complexes is reasonably explained in terms of this theory. In addition, the process leads to a mechanism to drive an electron in a lower to a higher-energy state, thus providing a mechanism for the ejection of the electron to a nearby molecule (chlorophyll) or into the environment. The observed lifetimes of the electronically excited states are in accord/agreement with the investigations of Sundstr?m et al. and are in the range of pico-seconds and less. The change in electronic charge distribution in internuclear space as the system undergoes an electronic transition to a higher-energy state could, under appropriate physical conditions, lead to oscillating dipoles capable of transmitting energy from the carotenoid-chlorophylls chromophore to the reaction center by sending an electromagnetic wave (a photon) which provides a novel new mechanism for energy production. In the simplest version of the F?rster?CDexter theory, the excitation energy of a donor is transferred to an acceptor and then de-excited to the ground state by fluorescence with no electron being transferred. In the process proposed herein, charge and energy both are transferred from donor to acceptor which can further de-excite by fluorescence. The charge transfer time scale involving an actual transfer of electron is in the pico-second range.     

Direct calculation of electron transfer parameters through constrained density functional theory

It is shown that constrained density functional theory (DFT) can be used to access diabatic potential energy surfaces in the Marcus theory of electron transfer, thus providing a means to directly calculate the driving force and the inner-sphere reorganization energy. We present in this report an analytic expression for the forces in constrained DFT and their implementation in geometry optimization, a prerequisite for the calculation of electron transfer parameters. The method is then applied to study the symmetric mixed-valence complex tetrathiafulvalene-diquinone radical anion, which is observed experimentally to be a Robin-Day class II compound but found by DFT to be in class III. Constrained DFT avoids this pitfall of over-delocalization and provides a way to find the charge-localized structure. In another application, driving forces and inner-sphere reorganization energies are calculated for the charge recombination (CR) reactions in formanilide-anthraquinone (FA-AQ) and ferrocene-formanilide-anthraquinone (Fc-FA-AQ). While the two compounds have similar reorganization energies, the driving force in FA-AQ is 1 eV larger than in Fc-FA-AQ, in agreement with experimental observations and supporting the experimental conclusion that the anomalously long-lived FA-AQ charge-separated state arises because the electron transfer is in the Marcus inverted region.     

The Rehm-Weller experiment in view of distant electron transfer

The driving‐force dependence of bimolecular fluorescence quenching by electron transfer in solution, the Rehm–Weller experiment, is revisited. One of the three long‐standing unsolved questions about the features of this experiment is carefully analysed here, that is, is there a diffusional plateau? New experimental quenching rates are compiled for a single electron donor, 2,5‐bis(dimethylamino)‐1,3‐benzenedicarbonitrile, and eighteen electron acceptors in acetonitrile. The data are analysed in the framework of differential encounter theory by using an extended version of the Marcus theory to model the intrinsic electron‐transfer step. Only by including the hydrodynamic effect and the solvent structure can the experimental findings be well modelled. The diffusional control region, the “plateau”, reveals the inherent distance dependence of the reaction, which is shown to be a general feature of electron transfer in solution.     

Conformational control of the Q(A) to Q(B) electron transfer in bacterial reaction centers: evidence for a frozen conformational landscape below -25 degrees C

The competition between the P(+)Q(A)(-) --> PQ(A) charge recombination (P, bacteriochlorophyll pair acting as primary photochemical electron donor) and the electron transfer to the secondary quinone acceptor Q(A)(-)Q(B) --> Q(A)Q(B)(-) (Q(A) and Q(B), primary and secondary electron accepting quinones) was investigated in chromatophores of Rb. capsulatus, varying the temperature down to -65 degrees C. The analysis of the flash-induced pattern for the formation of P(+)Q(A)Q(B)(-) shows that the diminished yield, when lowering the temperature, is not due to a homogeneous slowing of the rate constant k(AB) of the Q(A)(-)Q(B) --> Q(A)Q(B)(-) electron transfer but to a distribution of conformations that modulate the electron transfer rate over more than 3 orders of magnitude. This distribution appears "frozen", as no dynamic redistribution was observed over time ranges > 10 s (below -25 degrees C). The kinetic pattern was analyzed to estimate the shape of the distribution of k(AB), showing a bell-shaped band on the high rate side and a fraction of "blocked" reaction centers (RCs) with very slow k(AB). When the temperature is lowered, the high rate band moves to slower rate regions and the fraction of blocked RCs increases at the expense of the high rate band. The RCs that recombine from the P(+)Q(A)Q(B)(-) state appear temporarily converted to a state with rapid k(AB), indicating that the stabilized state described by Kleinfeld et al. (Biochemistry 1984, 23, 5780-5786) is still accessible at -60 degrees C.