Lycopene crystalloids exhibit singlet exciton fission in tomatoes.

Transient absorption studies conducted on in vitro lycopene aggregates, as well as on lycopene crystalloids inside tomato chromoplasts, reveal the appearance of a long-lived excited state, which we unambiguously identified as lycopene triplet. These triplet states must be generated by singlet exciton fission, which occurs from the lycopene 2Ag state. This is the first time the singlet fission process has ever been shown to occur in a biological material. We propose that the formation of carotenoid assemblies in chromoplasts may constitute a photoprotective process during chromoplast maturation, in addition to their function in signaling processes.

Modeling of ultrafast time-resolved fluorescence applied to a weakly coupled chromophore pair.

We present theory for calculating the third-order non-linear response function of a molecular aggregate in the weak inter-chromophore coupling regime. This approach is based on the perturbative expansion of the system evolution with respect to the resonance coupling, while the system-bath interaction is treated non-perturbatively by means of cumulant expansion. An explicit expression for the time-resolved fluorescence signal is then obtained. This allows us to investigate the ultrafast time-dependent Stokes shift, signatures of coherent dynamics, and the excitonic polaron formation in the excited state of the aggregate. Numerical simulations of the time-resolved fluorescence spectra of a pair of coupled molecules demonstrate these effects.

Light-harvesting processes in the dynamic photosynthetic antenna.

We present our perspective on the theoretical basis of light-harvesting within the photosynthetic membrane. Far from being a static structure, the photosynthetic membrane is a highly dynamic system, with protein mobility playing an important role in the damage/repair cycle of photosystem II (PSII), in balancing the input of energy between PSI and PSII, and in the photoprotection of PSII in response to a sudden excess of illumination. The concept of a photosynthetic antenna is illustrated and the state transition phenomenon is discussed as an example of purposeful antenna mobility. We discuss fluorescence recovery after photo-bleaching as a technique for visualising membrane mobility, before introducing light-induced grana membrane reorganisation as an integral part of the rapid photoprotective switch in plants. We then discuss current theoretical approaches to modelling the energy transfer dynamics of the PSII antenna: the atomistic models of intra-complex transfer and the coarse-grained approach to the inter-complex dynamics. Finally we discuss the future prospect of extending these methods, beyond the static picture of the membrane, to the dynamic PSII photosynthetic antenna.

Quantum mechanical calculations of xanthophyll-chlorophyll electronic coupling in the light-harvesting antenna of photosystem II of higher plants.

Light-harvesting by the xanthophylls in the antenna of photosystem II (PSII) is a very efficient process (with 80% of the absorbed energy being transfer to chlorophyll). However, the efficiencies of the individual xanthophylls vary considerably, with violaxanthin in LHCII contributing very little to light-harvesting. To investigate the origin of the variation we used Time Dependent Density Functional Theory to model the Coulombic interactions between the xanthophyll 1(1)B(u)(+) states and the chlorophyll Soret band states in the LHCII and CP29 antenna complexes. The results show that the central L1 and L2 binding sites in both complexes favored close cofacial associations between the bound xanthophylls and chlorophyll a, implying efficient energy transfer, consistent with previously reported experimental evidence. Additionally, we found that the peripheral V1 binding site in LHCII did not favor close xanthophyll-chlorophyll associations, confirming observations that violaxanthin in LHCII is not an effective light-harvester. Finally, violaxanthin bound into the L2 site of the CP29 complex was found to be very strongly coupled to its neighboring chlorophylls.

Excitation energy transfer and quenching in a heterodimer: applications to the carotenoid-phthalocyanine dyads.

The dynamics of a molecular heterodimer composed of a long-lived excitation donor and a short-lived acceptor (quencher) is examined. In order to consider various dynamical regimes without any restrictions on the system parameters, the energy transfer is modeled employing the hierarchical equations of motion, while the relaxation to the ground state is treated by assuming a phenomenological spontaneous nonradiative decay rate. Time scales of the resulting two-exponential evolution are investigated as functions of the energy gap and the resonance coupling between the monomeric constituents of the dimer. Relevance of the present analysis to the recent experimental findings on artificial carotenoid-phthalocyanine dyads is discussed. By examining the first two time scales of the reported time-resolved spectra, it is shown that upon the increase of carotenoid conjugation length its first excited state approaches the first excited state of phthalocyanine from above, thereby inducing a remarkable quenching. The proposed model also provides a unified treatment of quenching in the regimes previously distinguished as energy transfer and excitonic state formation.

Modeling of fluorescence quenching by lutein in the plant light-harvesting complex LHCII.

Photoprotective non-photochemical quenching (NPQ) in higher plants is the result of the formation of energy quenching traps in the light-harvesting antenna of photosystem II (PSII). It has been proposed that this quenching trap is a lutein molecule closely associated with the chlorophyll terminal emitter of the major light-harvesting complex LHCII. We have used a combination of time-dependent density functional theory (TD-DFT) and the semiempirical MNDO-CAS-CI method to model the chlorophyll-lutein energy transfer dynamics of the highly quenched crystal structure of LHCII. Our calculations reveal that the incoherent "hopping" of energy from Chla612 to the short-lived, dipole forbidden 2(1)A(g)(-) state of lutein620 accounts for the strong fluorescence quenching observed in these crystals. This adds weight to the argument that the same dissipative pathway is responsible for in vivo NPQ.

Quantum coherence and disorder-specific effects in simulations of 2D optical spectra of one-dimensional J-aggregates.

Time-resolved two-dimensional photon-echo (2D PE) spectra of linear J-aggregates containing four molecules per unit cell--a model system for a concentrated water solution of the pseudo-isocyanine dye--is theoretically considered. Analysis of a single unit cell and the full-sized aggregate is consistently carried out. Spectral features of a single unit cell are sorted out by assigning them to specific double-sided Feynman diagrams. Two different approaches of disorder are employed, the resulting differences in the spectra are discussed, and the conditions for their utilization are described. Special attention is paid to quantum coherence dynamics of a single unit cell and the full-sized J-aggregate. Possibilities of probing quantum coherences, as well as performing quantum control via two-color 2D PE spectroscopy, are discussed.

Exciton dynamics in semiconducting carbon nanotubes.

We report a femtosecond transient absorption spectroscopic study on the (6, 5) single-walled carbon nanotubes and the (7, 5) inner tubes of a dominant double-walled carbon nanotube species. We found that the dynamics of exciton relaxation probed at the first transition-allowed state (E(11)) of a given tube type exhibits a markedly slower decay when the second transition-allowed state (E(22)) is excited than that measured by exciting its first transition-allowed state (E(11)). A linear intensity dependence of the maximal amplitude of the transient absorption signal is found for the E(22) excitation, whereas the corresponding amplitude scales linearly with the square root of the E(11) excitation intensity. Theoretical modeling of these experimental findings was performed by developing a continuum model and a stochastic model with explicit consideration of the annihilation of coherent excitons. Our detailed numerical simulations show that both models can reproduce reasonably well the initial portion of decay kinetics measured upon the E(22) and E(11) excitation of the chosen tube species, but the stochastic model gives qualitatively better agreement with the intensity dependence observed experimentally than those obtained with the continuum model.

A theoretical investigation of the photophysical consequences of major plant light-harvesting complex aggregation within the photosynthetic membrane.

Spectroscopic measurements of Arabidopsis leaves have shown that the energy-dependent component of non-photochemical quenching (NPQ), known as qE, is associated with an absorption change at 535 nm (ΔA(535)). Identical measurements on the zeaxanthin-deficient mutant npq1 reveal a similar spectroscopic signature at 525 nm (ΔA(525)). We investigated whether these red-shifts may arise from excitonic interactions among homodimers of xanthophylls, zeaxanthin, and violaxanthin, bound at the peripheral V1 binding site on adjacent light-harvesting complex II (LHCII) trimers. Estimates of the relative geometries of these pigment pairs were obtained from the structure of LHCII. The excitonic couplings of zeaxanthin and violaxanthin dimers were probed using the time-dependent density functional theory method (TD-DFT). Calculations indicated that dimers formed between zeaxanthin or violaxanthin molecules using the published LHCII structure resulted in absorption blue shifts, typical of an H-type (parallel) geometry. In contrast, if the volume of the LHCII structure was modified to reflect the change in membrane thickness that occurs upon ΔpH formation, then both zeaxanthin and violaxanthin dimers adopted a J-type (collinear) geometry, and the resulting spectral shift was to the red region. The magnitudes of these predicted red-shifts are in good agreement with the experimental magnitudes. We therefore conclude that the observed xanthophyll red-shift results from the combination of both LHCII aggregation and changes in membrane thickness during qE. ΔA(535) may therefore be considered a "marker of aggregation" between LHCII trimers upon qE formation.

Static and dynamic protein impact on electronic properties of light-harvesting complex LH2.

A comparative analysis of the temperature dependence of the absorption spectra of the LH2 complexes from different species of photosynthetic bacteria, i.e., Rhodobacter sphaeroides, Rhodoblastus acidophilus, and Phaeospirillum molischianum, was performed in the temperature range from 4 to 300 K. Qualitatively, the temperature dependence is similar for all of the species studied. The spectral bandwidths of both B800 and B850 bands increases with temperature while the band positions shift in opposite directions: the B800 band shifts slightly to the red while the B850 band to the blue. These results were analyzed using the modified Redfield theory based on the exciton model. The main conclusion drawn from the analysis was that the spectral density function (SDF) is the main factor underlying the strength of the temperature dependence of the bandwidths for the B800 and B850 electronic transitions, while the bandwidths themselves are defined by the corresponding inhomogeneous distribution function (IDF). Slight variation of the slope of the temperature dependence of the bandwidths between species can be attributed to the changes of the values of the reorganization energies and characteristic frequencies determining the SDF. To explain the shift of the B850 band position with temperature, which is unusual for the conventional exciton model, a temperature dependence of the IDF must be postulated. This dependence can be achieved within the framework of the modified (dichotomous) exciton model. The slope of the temperature dependence of the B850 bandwidth is then defined by the value of the reorganization energy and by the difference between the transition energies of the dichotomous states of the pigment molecules. The equilibration factor between these dichotomous states mainly determines the temperature dependence of the peak shift.

Solvation effect of bacteriochlorophyll excitons in light-harvesting complex LH2.

We have characterized the influence of the protein environment on the spectral properties of the bacteriochlorophyll (Bchl) molecules of the peripheral light-harvesting (or LH2) complex from Rhodobacter sphaeroides. The spectral density functions of the pigments responsible for the 800 and 850 nm electronic transitions were determined from the temperature dependence of the Bchl absorption spectra in different environments (detergent micelles and native membranes). The spectral density function is virtually independent of the hydrophobic support that the protein experiences. The reorganization energy for the B850 Bchls is 220 cm(-1), which is almost twice that of the B800 Bchls, and its Huang-Rhys factor reaches 8.4. Around the transition point temperature, and at higher temperatures, both the static spectral inhomogeneity and the resonance interactions become temperature-dependent. The inhomogeneous distribution function of the transitions exhibits less temperature dependence when LH2 is embedded in membranes, suggesting that the lipid phase protects the protein. However, the temperature dependence of the fluorescence spectra of LH2 cannot be fitted using the same parameters determined from the analysis of the absorption spectra. Correct fitting requires the lowest exciton states to be additionally shifted to the red, suggesting the reorganization of the exciton spectrum.

Temperature broadening of LH2 absorption in glycerol solution.

In order to determine the relationship between the pigment-protein and the pigment-pigment interactions, the measurements of absorption spectra of the peripheral light-harvesting complex LH2 from the purple bacteria Rhodobacter sphaeroides solvated in glycerol/buffer solution were carried out in a wide temperature range, from 4 to 250 K. The SDFs used for simulating the temperature dependence of B800 and B850 bands were determined in a parametric form. To fit experimental spectra the overall exciton-phonon coupling had to be assumed to be weak for B850 (lambda/2V approximately 0.3, where lambda is the reorganization energy and V is the nearest-neighbor dipole-dipole coupling for bacteriochlorophylls). At physiological temperatures the intermediate nuclear bath dynamics compares with the magnitude of energy gap fluctuations. Slower dynamics with kappa approximately 0.39, where kappa is the ratio of the nuclear relaxation rate and the line width parameter, determines the spectral shape of B850 whilst faster modulations characterize B800 (kappa approximately 2.39). The static disorder for the B800 band is relatively high with the characteristic value of the inhomogeneous bandwidth Gamma(inh) approximately 120 cm-1, while for the B850 band this value is almost equal to the dipole-dipole coupling strength (Gamma(inh) approximately 360 cm-1). It has been found that the LH2 absorption spectrum is likely to be influenced by the temperature dependence of the dielectric constant of the solution in the high temperature range, when the glycerol/buffer solution is in the liquid state.

Spectral inhomogeneity of photosystem I and its influence on excitation equilibration and trapping in the cyanobacterium Synechocystis sp. PCC6803 at 77 K.

Ultrafast transient absorption spectroscopy was used to probe excitation energy transfer and trapping at 77 K in the photosystem I (PSI) core antenna from the cyanobacterium Synechocystis sp. PCC 6803. Excitation of the bulk antenna at 670 and 680 nm induces a subpicosecond energy transfer process that populates the Chl a spectral form at 685--687 nm within few transfer steps (300--400 fs). On a picosecond time scale equilibration with the longest-wavelength absorbing pigments occurs within 4-6 ps, slightly slower than at room temperature. At low temperatures in the absence of uphill energy transfer the energy equilibration processes involve low-energy shifted chlorophyll spectral forms of the bulk antenna participating in a 30--50-ps process of photochemical trapping of the excitation by P(700). These spectral forms might originate from clustered pigments in the core antenna and coupled chlorophylls of the reaction center. Part of the excitation is trapped on a pool of the longest-wavelength absorbing pigments serving as deep traps at 77 K. Transient hole burning of the ground-state absorption of the PSI with excitation at 710 and 720 nm indicates heterogeneity of the red pigment absorption band with two broad homogeneous transitions at 708 nm and 714 nm (full-width at half-maximum (fwhm) approximately 200--300 cm(-1)). The origin of these two bands is attributed to the presence of two chlorophyll dimers, while the appearance of the early time bleaching bands at 683 nm and 678 nm under excitation into the red side of the absorption spectrum (>690 nm) can be explained by borrowing of the dipole strength by the ground-state absorption of the chlorophyll a monomers from the excited-state absorption of the dimeric red pigments.

Singlet-singlet annihilation kinetics in aggregates and trimers of LHCII.

Singlet-singlet annihilation experiments have been performed on trimeric and aggregated light-harvesting complex II (LHCII) using picosecond spectroscopy to study spatial equilibration times in LHCII preparations, complementing the large amount of data on spectral equilibration available in literature. The annihilation kinetics for trimers can well be described by a statistical approach, and an annihilation rate of (24 ps)(-1) is obtained. In contrast, the annihilation kinetics for aggregates can well be described by a kinetic approach over many hundreds of picoseconds, and it is shown that there is no clear distinction between inter- and intratrimer transfer of excitation energy. With this approach, an annihilation rate of (16 ps)(-1) is obtained after normalization of the annihilation rate per trimer. It is shown that the spatial equilibration in trimeric LHCII between chlorophyll a molecules occurs on a time scale that is an order of magnitude longer than in Photosystem I-core, after correcting for the different number of chlorophyll a molecules in both systems. The slow transfer in LHCII is possibly an important factor in determining excitation trapping in Photosystem II, because it contributes significantly to the overall trapping time.

Electrical-to-mechanical coupling in purple membranes: membrane as electrostrictive medium.

In this paper, we present acousto-electrical measurements performed on dry films of purple membranes (PM) of Halobacterium salinarium. The purpose of these measurements is to determine the relation between mechanical and electrical phenomena in bacteriorhodopsin and to define the role of the protein in the proton transfer process. Electrical-to-mechanical coupling in PMs manifests itself as direct and inverse piezoelectric effects. Measurements performed on the samples with different degrees of PM orientation and at various values of the externally applied cross-membrane electric field indicate that piezoelectric phenomena in PMs arise from the electric asymmetry of the membranes, i.e., they originate from electrostriction. Experiments with samples made of oriented PMs allow estimation of the value of the intrinsic cross-membrane electric field, which is approximately 10(8) V/m. A hypothetical model of PM is presented where the electrical-to-mechanical coupling is suggested to be the main driving force for the proton translocation against the Coulomb forces acting in the membrane.

Generation of fluorescence quenchers from the triplet states of chlorophylls in the major light-harvesting complex II from green plants.

Laser flash-induced changes of the fluorescence yield were studied in aggregates of light-harvesting complex II (LHCII) on a time scale ranging from microseconds to seconds. Carotenoid (Car) and chlorophyll (Chl) triplet states, decaying with lifetimes of several microseconds and hundreds of microseconds, respectively, are responsible for initial light-induced fluorescence quenching via singlet-triplet annihilation. In addition, at times ranging from milliseconds to seconds, a slow decay of the light-induced fluorescence quenching can be observed, indicating the presence of additional quenchers generated by the laser. The generation of the quenchers is found to be sensitive to the presence of oxygen. It is proposed that long-lived fluorescence quenchers can be generated from Chl triplets that are not transferred to Car molecules. The quenchers could be Chl cations or other radicals that are produced directly from Chl triplets or via Chl triplet-sensitized singlet oxygen. Decay of the quenchers takes place on a millisecond to second time scale. The decay is slowed by a few orders of magnitude at 77 K indicating that structural changes or migration-limited processes are involved in the recovery. Fluorescence quenching is not observed for trimers, which is explained by a reduction of the quenching domain size compared to that of aggregates. This type of fluorescence quenching can operate under very high light intensities when Chl triplets start to accumulate in the light-harvesting antenna.

Energy migration in the light-harvesting antenna of the photosynthetic bacterium Rhodospirillum rubrum studied by time-resolved excitation annihilation at 77 K.

The intensity dependence of picosecond kinetics in the light-harvesting antenna of the photosynthetic bacterium Rhodospirillum rubrum is studied at 77 K. By changing either the average excitation intensity or the pulse intensity we have been able to discriminate singlet-singlet and singlet-triplet annihilation. It is shown that the kinetics of both annihilation types are well characterized by the concept of percolative excitation dynamics leading to the time-dependent annihilation rates. The time dependence of these two types of annihilation rates is qualitatively different, whereas the dependencies can be related through the same adjustable parameter-a spectral dimension of fractal-like structures. The theoretical dependencies give a good fit to the experimental kinetics if the spectral dimension is equal to 1.5 and the overall singlet-singlet annihilation rate is close to the value obtained at room temperature. The percolative transfer is a consequence of spectral inhomogeneous broadening. The effect is more pronounced at lower temperatures because of the narrowing of homogeneous spectra.

A perturbed two-level model for exciton trapping in small photosynthetic systems.

The study of exciton trapping in photosynthetic systems provides significant information about migration kinetics within the light harvesting antenna (LHA) and the reaction center (RC). We discuss two random walk models for systems with weakly coupled pigments, with a focus on the application to small systems (10-40 pigments/RC). Details of the exciton transfer to and from the RC are taken into consideration, as well as migration within the LHA and quenching in the RC. The first model is obtained by adapting earlier local trap models for application to small systems. The exciton lifetime is approximated by the sum of three contributions related to migration in the LHA, trapping by the RC, and quenching within the RC. The second model is more suitable for small systems and regards the finite rate of migration within the LHA as a perturbation of the simplified model, where the LHA and the RC are each represented by a single pigment level. In this approximation, the exciton lifetime is the sum of a migration component and a single nonlinear expression for the trapping and quenching of the excitons. Numerical simulations demonstrate that both models provide accurate estimates of the exciton lifetime in the intermediate range of 20-50 sites/RC. In combination, they cover the entire range of very small to very large photosynthetic systems. Although initially intended for regular LHA lattices, the models can also be applied to less regular systems. This becomes essential as more details of the structure of these systems become available. Analysis with these models indicates that the excited state decay in LH1 is limited by the average rate at which excitons transfer to the RC from neighboring sites in the LHA. By comparing this to the average rate of transfer within the LHA, various structural models that have been proposed for the LH1 core antenna are discussed.

Nonlinear annihilation of excitations in photosynthetic systems.

The theory of the singlet-singlet annihilation in quasi-homogeneous photosynthetic antenna systems is developed further. In the new model, the following important contributions are taken into account: 1) the finite excitation pulse duration, 2) the occupation of higher excited states during the annihilation, 3) excitation correlation effects, and 4) the effect of local heating. The main emphasis is concentrated on the analysis of pump-probe kinetic measurements demonstrating the first two above possible contributions. The difference with the results obtained from low-intensity fluorescence kinetic measurements is highlighted. The experimental data with picosecond time resolution obtained for the photosynthetic bacterium Rhodospirillum rubrum at room temperature are discussed on the basis of this theory.

Description of energy migration and trapping in photosystem I by a model with two distance scaling parameters.

The energy transfer and trapping kinetics in the core antenna of Photosystem I are described in a new model in which the distance between the core antenna chlorophylls and P700 is proposed to be considerably longer than the distance between the chlorophylls within the antenna. Structurally, the model describes the Photosystem I core antenna as a regular sphere around P700, while energetically it consists of three levels representing the bulk antenna, P700 and the red-shifted antenna pigments absorbing at longer wavelength than P700, respectively. It is shown that the model explains experimental results obtained from the Photosystem I complex of the cyanobacterium Synechococcus sp. (A.R. Holzwarth, G. Schatz, H Brock, and E. Bittersman (1993) Biophys. J. 64: 1813-1826) quite well, and that no unrealistic charge separation rate and organization of the long-wavelength pigments has to be assumed. We suggest that excitation energy transfer and trapping in Photosystem I should be described as a 'transfer-to-the-trap'-limited process.