Dynamics Of Photosystem Research Articles

Energy Transfer and Radical-Pair Dynamics in Photosystem I with Different Red Chlorophyll a Pigments.

We establish a general kinetic scheme for the energy transfer and radical-pair dynamics in photosystem I (PSI) of Chlamydomonas reinhardtii, Synechocystis PCC6803, Thermosynechococcus elongatus and Spirulina platensis grown under white-light conditions. With the help of simultaneous target analysis of transient-absorption data sets measured with two selective excitations, we resolved the spectral and kinetic properties of the different species present in PSI. WL-PSI can be described as a Bulk Chl a in equilibrium with a higher-energy Chl a, one or two Red Chl a and a reaction-center compartment (WL-RC). Three radical pairs (RPs) have been resolved with very similar properties in the four model organisms. The charge separation is virtually irreversible with a rate of ≈900 ns-1. The second rate, of RP1 → RP2, ranges from 70-90 ns-1 and the third rate, of RP2 → RP3, is ≈30 ns-1. Since RP1 and the Red Chl a are simultaneously present, resolving the RP1 properties is challenging. In Chlamydomonas reinhardtii, the excited WL-RC and Bulk Chl a compartments equilibrate with a lifetime of ≈0.28 ps, whereas the Red and the Bulk Chl a compartments equilibrate with a lifetime of ≈2.65 ps. We present a description of the thermodynamic properties of the model organisms at room temperature.

Energy flow in the Photosystem I supercomplex: Comparison of approximative theories with DM-HEOM

We analyze the exciton dynamics in Photosystem I from Thermosynechococcus elongatus using the distributed memory implementation of the hierarchical equation of motion (DM-HEOM) for the 96 Chlorophylls in the monomeric unit. The exciton-system parameters are taken from a first principles calculation. A comparison of the exact results with Förster rates and Markovian approximations allows one to validate the exciton transfer times within the complex and to identify deviations from approximative theories. We show the optical absorption, linear, and circular dichroism spectra obtained with DM-HEOM and compare them to experimental results.

Modeling of the redox state dynamics in photosystem II of Chlorella pyrenoidosa Chick cells and leaves of spinach and Arabidopsis thaliana from single flash-induced fluorescence quantum yield changes on the 100ns-10s time scale.

The time courses of the photosystem II (PSII) redox states were analyzed with a model scheme supposing a fraction of 11-25% semiquinone (with reduced [Formula: see text]) RCs in the dark. Patterns of single flash-induced transient fluorescence yield (SFITFY) measured for leaves (spinach and Arabidopsis (A.) thaliana) and the thermophilic alga Chlorella (C.) pyrenoidosa Chick (Steffen et al. Biochemistry 44:3123-3132, 2005; Belyaeva et al. Photosynth Res 98:105-119, 2008, Plant Physiol Biochem 77:49-59, 2014) were fitted with the PSII model. The simulations show that at high-light conditions the flash generated triplet carotenoid (3)Car(t) population is the main NPQ regulator decaying in the time interval of 6-8μs. So the SFITFY increase up to the maximum level [Formula: see text]/F 0 (at ~50μs) depends mainly on the flash energy. Transient electron redistributions on the RC redox cofactors were displayed to explain the SFITFY measured by weak light pulses during the PSII relaxation by electron transfer (ET) steps and coupled proton transfer on both the donor and the acceptor side of the PSII. The contribution of non-radiative charge recombination was taken into account. Analytical expressions for the laser flash, the (3)Car(t) decay and the work of the water-oxidizing complex (WOC) were used to improve the modeled P680(+) reduction by YZ in the state S 1 of the WOC. All parameter values were compared between spinach, A. thaliana leaves and C. pyrenoidosa alga cells and at different laser flash energies. ET from [Formula: see text] slower in alga as compared to leaf samples was elucidated by the dynamics of [Formula: see text] fractions to fit SFITFY data. Low membrane energization after the 10ns single turnover flash was modeled: the ∆Ψ(t) amplitude (20mV) is found to be about 5-fold smaller than under the continuous light induction; the time-independent lumen pHL, stroma pHS are fitted close to dark estimates. Depending on the flash energy used at 1.4, 4, 100% the pHS in stroma is fitted to 7.3, 7.4, and 7.7, respectively. The biggest ∆pH difference between stroma and lumen was found to be 1.2, thus pH- dependent NPQ was not considered.

Excitation dynamics in Photosystem I from Chlamydomonas reinhardtii. Comparative studies of isolated complexes and whole cells

Identical time-resolved fluorescence measurements with ~3.5-ps resolution were performed for three types of PSI preparations from the green alga, Chlamydomonas reinhardtii: isolated PSI cores, isolated PSI–LHCI complexes and PSI–LHCI complexes in whole living cells. Fluorescence decay in these types of PSI preparations has been previously investigated but never under the same experimental conditions. As a result we present consistent picture of excitation dynamics in algal PSI. Temporal evolution of fluorescence spectra can be generally described by three decay components with similar lifetimes in all samples (6–8ps, 25–30ps, 166–314ps). In the PSI cores, the fluorescence decay is dominated by the two fastest components (~90%), which can be assigned to excitation energy trapping in the reaction center by reversible primary charge separation. Excitation dynamics in the PSI–LHCI preparations is more complex because of the energy transfer between the LHCI antenna system and the core. The average trapping time of excitations created in the well coupled LHCI antenna system is about 12–15ps longer than excitations formed in the PSI core antenna. Excitation dynamics in PSI–LHCI complexes in whole living cells is very similar to that observed in isolated complexes. Our data support the view that chlorophylls responsible for the long-wavelength emission are located mostly in LHCI. We also compared in detail our results with the literature data obtained for plant PSI.

Cofactors Involved in Light-Driven Charge Separation in Photosystem I Identified by Subpicosecond Infrared Spectroscopy

Photosystem I is one of the key players in the conversion of solar energy into chemical energy. While the chlorophyll dimer P(700) has long been identified as the primary electron donor, the components involved in the primary charge separation process in PSI remain undetermined. Here, we have studied the charge separation dynamics in Photosystem I trimers from Synechococcus elongatus by femtosecond vis-pump/mid-infrared-probe spectroscopy upon excitation at 700, 710, and 715 nm. Because of the high specificity of the infrared region for the redox state and small differences in the molecular structure of pigments, we were able to clearly identify specific marker bands indicating chlorophyll (Chl) oxidation. Magnitudes of chlorophyll cation signals are observed to increase faster than the time resolution of the experiment (~0.2 ps) upon both excitation conditions: 700 nm and selective red excitation. Two models, involving either ultrafast charge separation or charge transfer character of the red pigments in PSI, are discussed to explain this observation. A further increase in the magnitudes of cation signals on a subpicosecond time scale (0.8-1 ps) indicates the formation of the primary radical pair. Evolution in the cation region with time constants of 7 and 40 ps reveals the formation of the secondary radical pair, involving a secondary electron donor. Modeling of the data allows us to extract the spectra of the two radical pairs, which have IR signatures consistent with A+A₀- and P₇₀₀+A₁-. We conclude that the cofactor chlorophyll A acts as the primary donor in PSI. The existence of an equilibrium between the two radical pairs we interpret as concerted hole/electron transfer between the pairs of electron donors and acceptors, until after 40 ps, relaxation leads to a full population of the P₇₀₀+A₁. radical pair.

Flash-Induced Structural Dynamics in Photosystem II Membrane Fragments of Green Plants

Time-resolved quasielastic neutron scattering with laser excitation is a promising novel pump-probe approach, which opens up new perspectives for the study of protein-membrane dynamics in specific functional states of even complex systems. This is demonstrated here for the case of photosystem II membrane fragments with inhibited electron transfer. In contrast to the case of the model system bacteriorhodopsin, a transient reduction of the dynamics is observed approximately 160 micros after the actinic laser flash. This effect is the first observation of a modulated structural dynamics in photosystem II membrane fragments.

Light-Induced Dynamics in Photosystem I Electron Transfer

Protein dynamics are likely to play important, regulatory roles in many aspects of photosynthetic electron transfer, but a detailed description of these coupled protein conformational changes has been unavailable. In oxygenic photosynthesis, photosystem I catalyzes the light-driven oxidation of plastocyanin or cytochrome c and the reduction of ferredoxin. A chlorophyll (chl) a/a′ heterodimer, P700, is the secondary electron donor, and the two P700 chl, are designated PA and PB. We used specific chl isotopic labeling and reaction-induced Fourier-transform infrared spectroscopy to assign chl keto vibrational bands to PA and PB. In the cyanobacterium, Synechocystis sp. PCC 6803, the chl keto carbon was labeled from 13C-labeled glutamate, and the chl keto oxygen was labeled from 18O2. These isotope-based assignments provide new information concerning the structure of PA+, which is found to give rise to two chl keto vibrational bands, with frequencies at 1653 and 1687cm−1. In contrast, PA gives rise to one chl keto band at 1638cm−1. The observation of two PA+ keto frequencies is consistent with a protein relaxation-induced distribution in PA+ hydrogen bonding. These results suggest a light-induced conformational change in photosystem I, which may regulate the oxidation of soluble electron donors and other electron-transfer reactions. This study provides unique information concerning the role of protein dynamics in oxygenic photosynthesis.

A brief introduction of Kimiyuki Satoh

In this Special Issue of Photosynthesis Research (Structure, Function, and Dynamics of Photosystem II) in honor of Kimiyuki Satoh and Thomas J. Wydrzynski, we present here a brief introduction to the scientific career and achievements of Kimiyuki Satoh, a great scientist with numerous important contributions in photosynthesis research, especially in the field of photosystem II.

The effect of hydration on protein flexibility in photosystem II of green plants studied by quasielastic neutron scattering

The effect of hydration on protein dynamics in photosystem II (PS II) membrane fragments from spinach has been investigated by using the method of quasielastic neutron scattering (QENS) at room temperature. The QENS data obtained indicate that the protein dynamics is strongly dependent on the extent of hydration. In particular, the hydration-induced activation of localized diffusive protein motions and QA- reoxidation by QB in PS II appear to be correlated in their onset at a hydration value of about 45% relative humidity (r.h.). These findings underline the crucial functional relevance of localized diffusive protein motions on the picosecond-timescale for the reactions of light-induced photosynthetic water splitting under formation of plastoquinol and molecular oxygen in PS II of green plants.

Temperature- and Hydration-Dependent Protein Dynamics in Photosystem II of Green Plants Studied by Quasielastic Neutron Scattering

Protein dynamics in hydrated and vacuum-dried photosystem II (PS II) membrane fragments from spinach has been investigated by quasielastic neutron scattering (QENS) in the temperature range between 5 and 300 K. Three distinct temperature ranges can be clearly distinguished by active type(s) of protein dynamics: (A) At low temperatures (T < 120 K), the protein dynamics of both dry and hydrated PS II is characterized by harmonic vibrational motions. (B) In the intermediate temperature range (120 < T < 240 K), the total mean square displacement <u2>total slightly deviates from the predicted linear behavior. The QENS data indicate that this deviation, which is virtually independent of the extent of hydration, is due to a partial onset of diffusive protein motions. (C) At temperatures above 240 K, the protein flexibility drastically changes because of the onset of diffusive (large-amplitude) protein motions. This dynamical transition is clearly hydration-dependent since it is strongly suppressed in dry PS II. The thermally activated onset of protein flexibility as monitored by QENS is found to be strictly correlated with the temperature-dependent increase of the electron transport efficiency from Q(A)(-) to QB (Garbers et al. (1998) Biochemistry 37, 11399-11404). Analogously, the freezing of protein mobility by dehydration in dry PS II appears to be responsible for the blockage of Q(A)(-) reoxidation by Q(B) at hydration values lower than 45% r.h. (Kaminskaya et al. (2003) Biochemistry 42, 8119-8132). Similar effects were observed for reactions of the water-oxidizing complex as outlined in the Discussion section.

Late Assembly Steps and Dynamics of the Cyanobacterial Photosystem I

The dynamics of photosystem I assembly in cyanobacteria have been addressed using in vivo pulse-chase labeling of Synechocystis sp. PCC 6803 proteins in combination with blue native polyacrylamide gel electrophoresis. The analyses indicate the existence of three different monomeric photosystem I complexes and also the high stability of photosystem I trimers. We show that in addition to a complete photosystem I monomer, containing all 11 subunits, we detected a PsaK-less monomer and a short-lived PsaL/PsaK-less complex. The latter two monomers were missing in the ycf37 mutant of Synechocystis sp. PCC 6803 that accumulates also less trimers. Pulse-chase experiments suggest that the three monomeric complexes have different functions in the biogenesis of the trimer. Based on these findings we propose a model where PsaK is incorporated in the latest step of photosystem I assembly. The PsaK-less photosystem I monomer may represent an intermediate complex that is important for the exchange of the two PsaK variants during high light acclimation. Implications of the presented data with respect to Ycf37 function are discussed.

Replacement of the methionine axial ligand to the primary electron acceptor A 0 slows the A 0− reoxidation dynamics in Photosystem I

The recent crystal structure of photosystem I (PSI) from Thermosynechococcus elongatus shows two nearly symmetric branches of electron transfer cofactors including the primary electron donor, P 700, and a sequence of electron acceptors, A, A 0 and A 1, bound to the PsaA and PsaB heterodimer. The central magnesium atoms of each of the putative primary electron acceptor chlorophylls, A 0, are unusually coordinated by the sulfur atom of methionine 688 of PsaA and 668 of PsaB, respectively. We [Ramesh et al. (2004a) Biochemistry 43:1369–1375] have shown that the replacement of either methionine with histidine in the PSI of the unicellular green alga Chlamydomonas reinhardtii resulted in accumulation of A 0 − (in 300-ps time scale), suggesting that both the PsaA and PsaB branches are active. This is in contrast to cyanobacterial PSI where studies with methionine-to-leucine mutants show that electron transfer occurs predominantly along the PsaA branch. In this contribution we report that the change of methionine to either leucine or serine leads to a similar accumulation of A 0 − on both the PsaA and the PsaB branch of PSI from C. reinhardtii, as we reported earlier for histidine mutants. More importantly, we further demonstrate that for all the mutants under study, accumulation of A 0 − is transient, and that reoxidation of A 0 − occurs within 1–2 ns, two orders of magnitude slower than in wild type PSI, most likely via slow electron transfer to A 1. This illustrates an indispensable role of methionine as an axial ligand to the primary acceptor A 0 in optimizing the rate of charge stabilization in PSI. A simple energetic model for this reaction is proposed. Our findings support the model of equivalent electron transfer along both cofactor branches in Photosystem I.

Are the trapping dynamics in Photosystem II sensitive to Q A redox potential?

Removal of the manganese complex from photosystem II (PSII) causes a 145 meV shift in the redox potential of the plastoquinone Q A [Biochim. Biophys. Acta 1229 (1995) 193; Photosynthetica 27 (1–2) (1992) 89; Biochim. Biophys. Acta 1229 (1995) 202]. Time resolved single photon counting has been used to monitor the time dependent concentrations of the singlet states in intact and manganese depleted BBY-type core particles studied with Q A in a fully open state. Surprisingly, no difference is observed between the experimental results obtained from the intact and manganese depleted samples. We demonstrate that this could be indicative of a very deep primary radical pair trap formed in PSII core particles with Q A in a fully open state. Alternatively, static inhomogeneity or a possible time-dependence of the Q A redox potential via relaxations such as proton motions should be considered.

Excited-state dynamics in photosystem II: insights from the x-ray crystal structure.

The heart of oxygenic photosynthesis is photosystem II (PSII), a multisubunit protein complex that uses solar energy to drive the splitting of water and production of molecular oxygen. The effectiveness of the photochemical reaction center of PSII depends on the efficient transfer of excitation energy from the surrounding antenna chlorophylls. A kinetic model for PSII, based on the x-ray crystal structure coordinates of 37 antenna and reaction center pigment molecules, allows us to map the major energy transfer routes from the antenna chlorophylls to the reaction center chromophores. The model shows that energy transfer to the reaction center is slow compared with the rate of primary electron transport and depends on a few bridging chlorophyll molecules. This unexpected energetic isolation of the reaction center in PSII is similar to that found in the bacterial photosystem, conflicts with the established view of the photophysics of PSII, and may be a functional requirement for primary photochemistry in photosynthesis. In addition, the model predicts a value for the intrinsic photochemical rate constant that is 4 times that found in bacterial reaction centers.

Structural/functional dynamics of photosystem II

Roughly half of the photosystem II polypeptides are encoded by the nucleus while the reaction centre core components are chloroplast encoded. The nuclear encoded photosynthetic proteins are synthesised in the cytosol with N-terminal extensions that direct the proteins to the right compartment in the cell. Various biochemical and crystallographic studies of PSII have shown that the assembled PSII-complex in vivo is a dimer. Building a functional PSII thus not only involves targeting to the envelope and the thylakoid membrane as well as processing and assembly of monomers, but also dimerization and lateral migration. Recent data strongly suggest that the nuclear encoded, 6.1 kDa psbW-protein takes part in the dimerization. We have combined an experimental system in which nuclear encoded radiolabelled precursor proteins are incubated with isolated chloroplasts, with blue native PAGE. The blue native tris/tricine system resolves nine distinct bands of which three correspond to PSII dimers/supercomplexes. Using this combined assay, kinetic import/assembly studies show that all so far analysed thylakoid membrane proteins are found initially in the stroma lamellae fractions before migration to their proper locations. It was found that the time for the PsbW-protein, from envelope targeting to incorporation into a fully assembled dimeric PSII, is less than three minutes. This In Organello assay in combination with site directed mutagenesis has also proven to work as a tool to analyse the importance of specific amino acid residues with respect to the PSII dimerization. The PsbW-protein N-terminal domain is being analysed and will be discussed.

Protein Dynamics in Photosystem II Complexes of Green Plants Studied by Time-Resolved Hole-Burning

We have studied the protein dynamics of three subcore reaction-center complexes of photosystem II: the isolated reaction center (RC), the core antenna CP47, and the CP47 RC complex by means of time-resolved hole-burning in the red wing of their Qy-absorption bands. The dependence of the “effective” homogeneous line width on temperature T between 1.2 and 4.2 K suggests that optical dephasing in these proteins is determined by two-level systems (TLSs), as in doped organic glasses. By contrast, the increase of as a function of delay time td (between burning and probing the hole) from 10-5 to 105 s, caused by spectral diffusion (SD), differs from that in glasses and is characteristic for each complex. CP47 RC does not undergo any SD over 10 decades in time for T ≤ 4.2 K, i.e., = constant at a given temperature. Although CP47 and the RC do not show SD for td ≤ 1 s, they do for longer delay times, with increasing logarithmically with td. The onset and amount of SD appear to be correlated with the mass of the protein. We conclude that only slow motions, related to TLSs located at the more flexible surface of the protein (with a broad and continuous distribution of rates R < 1−3 Hz), contribute to SD at long delay times and that the whole protein, or a substantial part of it, is involved in SD. Fast, local fluctuations associated with a rigid, crystalline-like inner protein core are responsible for “pure” dephasing at short td.

The structure, function and dynamics of photosystem two

One of the greatest challenges in modern photosynthesis research is to elucidate fully the structural and functional properties of photosystem two (PSII). This water‐plasto‐quinone oxidoreductase is located in a membrane complex composed of more than 25 subunits. The primary and secondary structures of all known subunits which constitute the central core of PSII are reviewed. How these subunits interact with each other to produce the tertiary and quaternary structure of PSII in vivo is not fully understood. However, electron microscopy is helping to fill this gap in our knowledge both by single particle analysis and electron crystallography. These studies suggest that active PSII is dimeric, although the functional significance of this oligomeric state is not yet understood. Moreover, the elucidation of the structure of photosystem one (PSI) by X‐ray crystallography has revealed features which are likely to be relevant to PSII structure. It seems highly likely that the D1 protein with CP43 and D2 protein with CP47 (summing 11 transmembrane helices in each case) will have structural similarities to the organisation of PsaA and PsaB. It is likely that the turnover of the D1 protein is aided by the relatively easy removal of CP43 from this arrangement of the PSII core.

The structure, function and dynamics of photosystem two

The structure, function and dynamics of photosystem two

Excited state dynamics in photosystem I: effects of detergent and excitation wavelength

Femtosecond transient absorption spectroscopy has been used to investigate the energy transfer and trapping processes in both intact membranes and purified detergent-isolated particles from a photosystem II deletion mutant of the cyanobacterium Synechocystis sp. PCC 6803, which contains only the photosystem I reaction center. Processes with similar lifetimes and spectra are observed in both the membrane fragments and the detergent-isolated particles, suggesting little disruption of the core antenna resulting from the detergent treatment. For the detergent-isolated particles, three different excitation wavelengths were used to excite different distributions of pigments in the spectrally heterogeneous core antenna. Only two lifetimes of 2.7-4.3 ps and 24-28 ps, and a nondecaying component are required to describe all the data. The 24-28 ps component is associated with trapping. The trapping process gives rise to a nondecaying spectrum that is due to oxidation of the primary electron donor. The lifetimes and spectra associated with trapping and radical pair formation are independent of excitation wavelength, suggesting that trapping proceeds from an equilibrated excited state. The 2.7-4.3 ps component characterizes the evolution from the initially excited distribution of pigments to the equilibrated excited state distribution. The spectrum associated with the 2.7-4.3 ps component is therefore strongly excitation wavelength dependent. Comparison of the difference spectra associated with the spectrally equilibrated state and the radical pair state suggests that the pigments in the photosystem I core antenna display some degree of excitonic coupling.

Excitation transport and trapping on spectrally disordered lattices

It is widely assumed that the decay of fluorescence in photosynthetic systems can be described as a sum of exponential components and that the amplitude of each component is directly related to the absorption cross-section of the antenna pigments coupled to the fluorescing species. We present exact calculations of excited state decay in two-dimensional regular lattices of different geometries containing multiple spectral forms of antenna pigments. We illustrate by these calculations that there is no simple relation between the decay amplitudes (and resulting time-resolved excitation spectra) and the steady-state absorption spectra. Only in the limit that the electronic excitations reach a rapid equilibrium among all antenna spectral forms does the excitation spectrum depend uniquely on the spectral features of the array. Using the simulations in conjunction with our recent fluorescence studies, we examine excitation transport and trapping dynamics in photosystem I and the limitations imposed by the finite time resolution in single photon counting experiments. In particular, we show that rising components, associated with excitation transfer among different spectral forms, with lifetimes <20 ps would be undetected in a typical photon counting experiment.