RESUMO
Quantum coherences, observed as time-dependent beats in ultrafast spectroscopic experiments, arise when light-matter interactions prepare systems in superpositions of states with differing energy and fixed phase across the ensemble. Such coherences have been observed in photosynthetic systems following ultrafast laser excitation, but what these coherences imply about the underlying energy transfer dynamics remains subject to debate. Recent work showed that redox conditions tune vibronic coupling in the Fenna-Matthews-Olson (FMO) pigment-protein complex in green sulfur bacteria, raising the question of whether redox conditions may also affect the long-lived (>100 fs) quantum coherences observed in this complex. In this work, we perform ultrafast two-dimensional electronic spectroscopy measurements on the FMO complex under both oxidizing and reducing conditions. We observe that many excited-state coherences are exclusively present in reducing conditions and are absent or attenuated in oxidizing conditions. Reducing conditions mimic the natural conditions of the complex more closely. Further, the presence of these coherences correlates with the vibronic coupling that produces faster, more efficient energy transfer through the complex under reducing conditions. The growth of coherences across the waiting time and the number of beating frequencies across hundreds of wavenumbers in the power spectra suggest that the beats are excited-state coherences with a mostly vibrational character whose phase relationship is maintained through the energy transfer process. Our results suggest that excitonic energy transfer proceeds through a coherent mechanism in this complex and that the coherences may provide a tool to disentangle coherent relaxation from energy transfer driven by stochastic environmental fluctuations.
Assuntos
Transferência de Energia/fisiologia , Complexos de Proteínas Captadores de Luz/fisiologia , Fotossíntese/fisiologia , Proteínas de Bactérias/química , Luz , Complexos de Proteínas Captadores de Luz/metabolismo , Oxirredução , Complexo de Proteínas do Centro de Reação Fotossintética/fisiologia , Pigmentação , Teoria Quântica , Análise Espectral/métodos , VibraçãoRESUMO
Conversion of light energy into chemical energy through photosynthesis in the chloroplasts of photosynthetic organisms is essential for photoautotrophic growth, and non-photochemical quenching (NPQ) of excess light energy prevents the generation of reactive oxygen species and maintains efficient photosynthesis under high light. In the unicellular green alga Chlamydomonas reinhardtii, NPQ is activated as a photoprotective mechanism through wavelength-specific light signaling pathways mediated by the phototropin (blue light) and ultra-violet (UV) light photoreceptors, but the biological significance of photoprotection activation by light with different qualities remains poorly understood. Here, we demonstrate that NPQ-dependent photoprotection is activated more rapidly by UV than by visible light. We found that induction of gene expression and protein accumulation related to photoprotection was significantly faster and greater in magnitude under UV treatment compared with that under blue- or red-light treatment. Furthermore, the action spectrum of UV-dependent induction of photoprotective factors implied that C. reinhardtii senses relatively long-wavelength UV (including UV-A/B), whereas the model dicot plant Arabidopsis (Arabidopsis thaliana) preferentially senses relatively short-wavelength UV (mainly UV-B/C) for induction of photoprotective responses. Therefore, we hypothesize that C. reinhardtii developed a UV response distinct from that of land plants.
Assuntos
Arabidopsis/genética , Arabidopsis/fisiologia , Chlamydomonas reinhardtii/genética , Chlamydomonas reinhardtii/fisiologia , Complexos de Proteínas Captadores de Luz/fisiologia , Fotossíntese/fisiologia , Raios UltravioletaRESUMO
Photosystem II (PSII) is a large membrane protein complex performing primary charge separation in oxygenic photosynthesis. The biogenesis of PSII is a complicated process that involves a coordinated linking of assembly modules in a precise order. Each such module consists of one large chlorophyll (Chl)-binding protein, number of small membrane polypeptides, pigments and other cofactors. We isolated the CP47 antenna module from the cyanobacterium Synechocystis sp. PCC 6803 and found that it contains a 11-kDa protein encoded by the ssl2148 gene. This protein was named Psb35 and its presence in the CP47 module was confirmed by the isolation of FLAG-tagged version of Psb35. Using this pulldown assay, we showed that the Psb35 remains attached to CP47 after the integration of CP47 into PSII complexes. However, the isolated Psb35-PSIIs were enriched with auxiliary PSII assembly factors like Psb27, Psb28-1, Psb28-2 and RubA while they lacked the lumenal proteins stabilizing the PSII oxygen-evolving complex. In addition, the Psb35 co-purified with a large unique complex of CP47 and photosystem I trimer. The absence of Psb35 led to a lower accumulation and decreased stability of the CP47 antenna module and associated high-light-inducible proteins but did not change the growth rate of the cyanobacterium under the variety of light regimes. Nevertheless, in comparison with WT, the Psb35-less mutant showed an accelerated pigment bleaching during prolonged dark incubation. The results suggest an involvement of Psb35 in the life cycle of cyanobacterial Chl-binding proteins, especially CP47.
Assuntos
Proteínas de Bactérias/metabolismo , Complexos de Proteínas Captadores de Luz/metabolismo , Complexo de Proteína do Fotossistema II/metabolismo , Synechocystis/metabolismo , Proteínas de Bactérias/química , Proteínas de Bactérias/fisiologia , Proteínas de Bactérias/efeitos da radiação , Luz , Complexos de Proteínas Captadores de Luz/química , Complexos de Proteínas Captadores de Luz/fisiologia , Proteínas de Membrana/metabolismo , Proteínas de Membrana/fisiologia , Complexo de Proteína do Fotossistema II/química , Complexo de Proteína do Fotossistema II/fisiologia , Estrutura Terciária de Proteína , Synechocystis/efeitos da radiaçãoRESUMO
Phycobiliproteins (PBPs) are pigment proteins that comprise phycobilisomes (PBS), major light-harvesting antenna complexes of cyanobacteria and red algae. PBS core substructures are made up of allophycocyanins (APs), a subfamily of PBPs. Five paralogous AP subunits are encoded by the Far-Red Light Photoacclimation (FaRLiP) gene cluster, which is transcriptionally activated in cells grown in far-red light (FRL; λ = 700 to 800 nm). FaRLiP gene expression enables some terrestrial cyanobacteria to remodel their PBS and photosystems and perform oxygenic photosynthesis in far-red light (FRL). Paralogous AP genes encoding a putative, FRL-absorbing AP (FRL-AP) are also found in an operon associated with improved low-light growth (LL; < 50 µmol photons m-2 s-1) in some thermophilic Synechococcus spp., a phenomenon termed low-light photoacclimation (LoLiP). In this study, apc genes from FaRLiP and LoLiP gene clusters were heterologously expressed individually and in combinations in Escherichia coli. The resulting novel FRL-APs were characterized and identified as major contributors to the FRL absorbance observed in whole cells after FaRLiP and potentially LoLiP. Post-translational modifications of native FRL-APs from FaRLiP cyanobacterium, Leptolyngbya sp. strain JSC-1, were analyzed by mass spectrometry. The PBP complexes made in two FaRLiP organisms were compared, revealing strain-specific diversity in the FaRLiP responses of cyanobacteria. Through analyses of native and recombinant proteins, we improved our understanding of how different cyanobacterial strains utilize specialized APs to acclimate to FRL and LL. We discuss some insights into structural changes that may allow these APs to absorb longer light wavelengths than their visible-light-absorbing paralogs.
Assuntos
Cianobactérias/química , Luz , Ficocianina/química , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Complexos de Proteínas Captadores de Luz/química , Complexos de Proteínas Captadores de Luz/fisiologia , Complexo de Proteína do Fotossistema IRESUMO
Photosynthesis achieves near unity light-harvesting quantum efficiency yet it remains unknown whether there exists a fundamental organizing principle giving rise to robust light harvesting in the presence of dynamic light conditions and noisy physiological environments. Here, we present a noise-canceling network model that relates noisy physiological conditions, power conversion efficiency, and the resulting absorption spectra of photosynthetic organisms. Using light conditions in full solar exposure, light filtered by oxygenic phototrophs, and light filtered under seawater, we derived optimal absorption characteristics for efficient solar power conversion. We show how light-harvesting antennae can be tuned to maximize power conversion efficiency by minimizing excitation noise, thus providing a unified theoretical basis for the observed wavelength dependence of absorption in green plants, purple bacteria, and green sulfur bacteria.
Assuntos
Complexos de Proteínas Captadores de Luz/fisiologia , Fotossíntese , Plantas/metabolismo , Proteobactérias/metabolismo , Adsorção , Chlorobi , Transferência de Energia , Luz , Oxigênio , Energia SolarRESUMO
One of the major factors limiting biomass productivity in algae is the low thermodynamic efficiency of photosynthesis. The greatest thermodynamic inefficiencies in photosynthesis occur during the conversion of light into chemical energy. At full sunlight the light-harvesting antenna captures photons at a rate nearly 10 times faster than the rate-limiting step in photosynthetic electron transport. Excess captured energy is dissipated by non-productive pathways including the production of reactive oxygen species. Substantial improvements in photosynthetic efficiency have been achieved by reducing the optical cross-section of the light-harvesting antenna by selectively reducing chlorophyll b levels and peripheral light-harvesting complex subunits. Smaller light-harvesting antenna, however, may not exhibit optimal photosynthetic performance in low or fluctuating light environments. We describe a translational control system to dynamically adjust light-harvesting antenna sizes for enhanced photosynthetic performance. By expressing a chlorophyllide a oxygenase (CAO) gene having a 5' mRNA extension encoding a Nab1 translational repressor binding site in a CAO knockout line it was possible to continuously alter chlorophyll b levels and correspondingly light-harvesting antenna sizes by light-activated Nab1 repression of CAO expression as a function of growth light intensity. Significantly, algae having light-regulated antenna sizes had substantially higher photosynthetic rates and two-fold greater biomass productivity than the parental wild-type strains as well as near wild-type ability to carry out state transitions and non-photochemical quenching. These results have broad implications for enhanced algae and plant biomass productivity.
Assuntos
Clorófitas/metabolismo , Complexos de Proteínas Captadores de Luz/fisiologia , Fotossíntese , Biomassa , Clorofila/metabolismo , Clorófitas/crescimento & desenvolvimento , Clorófitas/fisiologia , Luz , Complexos de Proteínas Captadores de Luz/metabolismo , Fotossíntese/fisiologia , Plantas Geneticamente ModificadasRESUMO
In several systems, from plant's canopy to algal bioreactors, the decrease of the antenna size has been proposed as a strategy to increase the photosynthetic efficiency. However, still little is known about possible secondary effects of such modifications. This is particularly relevant because the modulation of the antenna size is one of the most important light acclimation responses in photosynthetic organisms. In our study, we used an Arabidopsis thaliana mutant (dLhcb2), which has a 60% decrease of Lhcb1 and Lhcb2, the two main components of the major Photosystem II antenna complex. We show that the mutant maintains the photosynthetic and photoprotective capacity of the Wild Type (WT) and adapts to different light conditions by remodelling its photosynthetic apparatus, but the regulatory mechanism differs from that of the WT. Surprisingly, it does not compensate for the decreased light-harvesting capacity by increasing other pigment-protein complexes. Instead, it lowers the ratio of the cytochrome b6 f and ATP synthase to the photosystems, regulating linear electron flow and maintaining the photosynthetic control at the level of these complexes as in the WT. We show that targeting the reduction of two specific antenna proteins, Lhcb1 and Lhcb2, represents a viable solution to obtain plants with a truncated antenna size, which still maintain the capacity to acclimate to different light conditions.
Assuntos
Arabidopsis/fisiologia , Complexos de Proteínas Captadores de Luz/fisiologia , Complexo de Proteína do Fotossistema II/fisiologia , Aclimatação , Arabidopsis/genética , Arabidopsis/efeitos da radiação , Clorofila/metabolismo , Complexo Citocromos b6f/metabolismo , Eletroforese em Gel de Poliacrilamida , Complexos de Proteínas Captadores de Luz/genética , Mutação , Complexo de Proteína do Fotossistema I/fisiologia , Tilacoides/metabolismoRESUMO
Light drives photosynthesis. In plants it is absorbed by light-harvesting antenna complexes associated with Photosystem I (PSI) and photosystem II (PSII). As PSI and PSII work in series, it is important that the excitation pressure on the two photosystems is balanced. When plants are exposed to illumination that overexcites PSII, a special pool of the major light-harvesting complex LHCII is phosphorylated and moves from PSII to PSI (state 2). If instead PSI is over-excited the LHCII complex is dephosphorylated and moves back to PSII (state 1). Recent findings have suggested that LHCII might also transfer energy to PSI in state 1. In this work we used a combination of biochemistry and (time-resolved) fluorescence spectroscopy to investigate the PSI antenna size in state 1 and state 2 for Arabidopsis thaliana. Our data shows that 0.7 ± 0.1 unphosphorylated LHCII trimers per PSI are present in the stroma lamellae of state-1 plants. Upon transition to state 2 the antenna size of PSI in the stroma membrane increases with phosphorylated LHCIIs to a total of 1.2 ± 0.1 LHCII trimers per PSI. Both phosphorylated and unphosphorylated LHCII function as highly efficient PSI antenna.
Assuntos
Arabidopsis/enzimologia , Complexos de Proteínas Captadores de Luz/fisiologia , Luz , Complexo de Proteína do Fotossistema I/efeitos da radiação , Arabidopsis/ultraestrutura , Digitonina/farmacologia , Transferência de Energia , Complexos de Proteínas Captadores de Luz/efeitos dos fármacos , Fosforilação , Complexo de Proteína do Fotossistema II/efeitos da radiação , Espectrometria de FluorescênciaRESUMO
Light-harvesting complexes (LHC) in photosynthetic organisms perform the major function of light absorption and energy transportation. Optical spectrum of LHC provides a detailed understanding of the molecular mechanisms involved in the excitation energy transfer (EET) processes, which has been widely studied. Here, we study how the geometric property of LHC in Rhodospirillum (Rs.) molischianum would affect its spectral characteristics and energy transfer process. By adopting the effective Hamiltonian and the dipole-dipole approximation, we calculate the exciton level structures for the LH2 ring and LH1 ring and the energy transfer time between different LHCs under various structural parameters and different rotational symmetries. Our numerical results show that the LHC's absorption peaks and the energy transfer time between different LHCs can be modified by changing the geometric configurations. Our study may be beneficial to the applications in designing highly efficient photovoltaic cell and other artificial photosynthetic systems.
Assuntos
Transferência de Energia/fisiologia , Complexos de Proteínas Captadores de Luz/fisiologia , Luz , Rhodospirillaceae/fisiologia , Conformação ProteicaRESUMO
Lichens and their algal partners are desiccation-tolerant organisms and as such survive after the complete loss of water. This trait is the consequence of several physiological, biochemical and structural features, including specific mechanisms dissipating excess light to avoid photooxidative stress. The maximum quantum yield of photosystem II (PSII; Fv /Fm ) is widely used as a sensitive indicator of photosynthetic performance and is calculated after complete relaxation in darkness of the fluorescence quenching associated with active light energy dissipation mechanisms. Unexpectedly, we observed that lichens and isolated chlorobionts (chlorophyte symbionts in lichen) maintained in darkness for several hours showed a strong decrease in the ratio Fv /Fm , which was reversible after re-illumination. We analyzed this dark-induced Fv /Fm decay in the chlorobiont Asterochloris erici through steady-state and fast-induction kinetics of chlorophyll a fluorescence and simultaneous P700 oxidation measurements. We found that the gradual decay of Fv /Fm in darkness was caused by reversible dark-induced inactivation of some PSII reaction centers that was accompanied by a decrease in the flux of electrons to PSI. Darkness induced the plastoquinone-reductase activity associated with chlororespiration and the phosphorylation of light harvesting complex (LHC). We propose that upon phosphorylation the LHC detaches from PSII, resulting in a decrease of exciton-trapping by PSII reaction centers and, consequently, an increased dissipation of light energy. This mechanism probably serves an ecophysiological function in lichens to prevent the damage at dawn or under strong fluctuating light conditions when lichens are in a hydrated state.
Assuntos
Clorofila A/química , Clorófitas/fisiologia , Escuridão , Fluorescência , Líquens/fisiologia , Complexo de Proteína do Fotossistema II/fisiologia , Complexos de Proteínas Captadores de Luz/fisiologia , FosforilaçãoRESUMO
Photosynthetic organisms require rapid and reversible down-regulation of light harvesting to avoid photodamage. Response to unpredictable light fluctuations is achieved by inducing energy-dependent quenching, qE, which is the major component of the process known as non-photochemical quenching (NPQ) of chlorophyll fluorescence. qE is controlled by the operation of the xanthophyll cycle and accumulation of specific types of proteins, upon thylakoid lumen acidification. The protein cofactors so far identified to modulate qE in photosynthetic eukaryotes are the photosystem II subunit S (PsbS) and light-harvesting complex stress-related (LHCSR/LHCX) proteins. A transition from LHCSR- to PsbS-dependent qE took place during the evolution of the Viridiplantae (also known as 'green lineage' organisms), such as green algae, mosses and vascular plants. Multiple studies showed that LHCSR and PsbS proteins have distinct functions in the mechanism of qE. LHCX(-like) proteins are closely related to LHCSR proteins and found in 'red lineage' organisms that contain secondary red plastids, such as diatoms. Although LHCX proteins appear to control qE in diatoms, their role in the mechanism remains poorly understood. Here, we present the current knowledge on the functions and evolution of these crucial proteins, which evolved in photosynthetic eukaryotes to optimise light harvesting.
Assuntos
Eucariotos/fisiologia , Oxigênio/química , Fotossíntese/fisiologia , Complexo de Proteína do Fotossistema II/fisiologia , Plantas/metabolismo , Bryopsida/fisiologia , Chlamydomonas/fisiologia , Clorofila/química , Luz , Complexos de Proteínas Captadores de Luz/fisiologia , Filogenia , Plastídeos/metabolismo , Xantofilas/químicaRESUMO
Non-photochemical quenching (NPQ) is a fast acting photoprotective response to high light stress triggered by over excitation of photosystem II. The mechanism for NPQ in the globally important diatom algae has been principally attributed to a xanthophyll cycle, analogous to the well-described qE quenching of higher plants. This study compared the short-term NPQ responses in two pennate, benthic diatom species cultured under identical conditions but which originate from unique light climates. Variable chlorophyll fluorescence was used to monitor photochemical and non-photochemical excitation energy dissipation during high light transitions; whereas whole cell steady state 77 K absorption and emission were used to measure high light elicited changes in the excited state landscapes of the thylakoid. The marine shoreline species Nitzschia curvilineata was found to have an antenna system capable of entering a deeply quenched, yet reversible state in response to high light, with NPQ being highly sensitive to dithiothreitol (a known inhibitor of the xanthophyll cycle). Conversely, the salt flat species Navicula sp. 110-1 exhibited a less robust NPQ that remained largely locked-in after the light stress was removed; however, a lower amplitude, but now highly reversible NPQ persisted in cells treated with dithiothreitol. Furthermore, dithiothreitol inhibition of NPQ had no functional effect on the ability of Navicula cells to balance PSII excitation/de-excitation. These different approaches for non-photochemical excitation energy dissipation are discussed in the context of native light climate.
Assuntos
Diatomáceas/fisiologia , Complexo de Proteína do Fotossistema II/fisiologia , Clorofila/metabolismo , Clorofila/fisiologia , Clima , Diatomáceas/efeitos da radiação , Transporte de Elétrons , Complexos de Proteínas Captadores de Luz/metabolismo , Complexos de Proteínas Captadores de Luz/fisiologia , Processos Fotoquímicos , Estresse Fisiológico , Luz Solar , Tilacoides/metabolismo , Tilacoides/fisiologiaRESUMO
In framework of the continuum electrostatics theory, the reorganization energies of the electron transfers QA--QB (fast phase), Bph--QA, P+-QA-, and P+-QB- in the photosynthetic bacterial reaction center have been calculated. The calculations were based on the static dielectric permittivity spatial distribution derived from the data on the electrogenesis, with the corresponding characteristic times relatively close to the reaction times of QA--QB (fast phase) and Bph--QA but much shorter than those times of the latter two recombination reactions. The calculated reorganization energies were reasonably close to the experimental estimates for QA--QB (fast phase) and Bph--QA but substantially lower than those of P+-QA- and P+-QB-. A higher effective dielectric permittivity contributes to this effect, but the dominant contribution is most probably made by a non-dielectric relaxation, especially for the P+-QB- recombination influenced by the proton transfer. This situation calls for reconsidering of the current electron transfer rate estimates.
Assuntos
Transporte de Elétrons/fisiologia , Complexos de Proteínas Captadores de Luz/fisiologia , Complexo de Proteínas do Centro de Reação Fotossintética/fisiologia , Quinonas/metabolismo , Rhodobacter sphaeroides/fisiologia , Complexos de Proteínas Captadores de Luz/química , Estrutura Molecular , Complexo de Proteínas do Centro de Reação Fotossintética/químicaRESUMO
The selective removal of B800 bacteriochlorophyll (BChl) a from light-harvesting complex 2 (LH2) in purple photosynthetic bacteria is a clue about elucidation of the mechanism for the transfer of energy from these pigments to B850 BChl a and their roles in the LH2 protein structure. We demonstrated that the kinetics of the removal of B800 BChl a from two representative LH2 proteins derived from Phaeospirillum molischianum and Rhodoblastus acidophilus differed significantly, in contrast to the calculated binding enthalpy. These results may be interpreted as changes in the local structure near B800 BChl a with respect to the geometries of the original crystal structures upon removal of B800 BChl a. Despite the difficulty of removing B800 BChl a from molischianum-LH2, we prepared the molischianum-LH2 protein lacking B800 BChl a by combination of two detergents, n-dodecyl ß-d-maltoside and n-octyl ß-d-glucoside, under acidic conditions. Spectral and atomic force microscopy analyses indicated that the absence of B800 BChl a had little effect on the local structure in the vicinity of B850 BChl a and the circular arrangement in this protein. These results suggest that the hydrophobic domain near B850 BChl a is rigid and plays a major role in the structural formation of molischianum-LH2.
Assuntos
Complexos de Proteínas Captadores de Luz/química , Proteobactérias/química , Bactérias/metabolismo , Proteínas de Bactérias/química , Bacterioclorofila A/metabolismo , Bacterioclorofilas/química , Transferência de Energia , Complexos de Proteínas Captadores de Luz/fisiologia , Fotossíntese , Conformação Proteica , Elementos Estruturais de Proteínas , Proteobactérias/metabolismoRESUMO
The circadian clock regulates many aspects of leaf gas supply and biochemical demand for CO2 , and is hypothesized to improve plant performance. Yet the extent to which the clock may regulate the efficiency of photosystem II (PSII) and photoprotective mechanisms such as heat dissipation is less explored. Based on measurements of chlorophyll a fluorescence, we estimated the maximum efficiency of PSII in light (Fv'/Fm') and heat dissipation by nonphotochemical quenching (NPQ). We further dissected total NPQ into its main components, qE (pH-dependent quenching), qT (state-transition quenching), and qI (quenching related to photoinhibition), in clock mutant genotypes of Arabidopsis thaliana, the cognate wild-type genotypes, and a panel of recombinant inbred lines expressing quantitative variation in clock period. Compared with mutants with altered clock function, we observed that wild-type genotypes with clock period lengths of approximately 24 hr had both higher levels of Fv'/Fm', indicative of improved PSII function, and reduced NPQ, suggestive of lower stress on PSII light harvesting complexes. In the recombinant inbred lines, genetic variances were significant for Fv'/Fm' and all 3 components of NPQ, with qE explaining the greatest proportion of NPQ. Bivariate tests of association and structural equation models of hierarchical trait relationships showed that quantitative clock variation was empirically associated with Fv'/Fm' and NPQ, with qE mediating the relationship with gas exchange. The results demonstrate significant segregating variation for all photoprotective components, and suggest the adaptive significance of the clock may partly derive from its regulation of the light reactions of photosynthesis and of photoprotective mechanisms.
Assuntos
Ritmo Circadiano , Complexo de Proteína do Fotossistema II/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/fisiologia , Clorofila A/metabolismo , Ritmo Circadiano/fisiologia , Luz , Complexos de Proteínas Captadores de Luz/metabolismo , Complexos de Proteínas Captadores de Luz/fisiologia , Complexo de Proteína do Fotossistema II/fisiologia , Folhas de Planta/metabolismo , Plantas Geneticamente ModificadasRESUMO
The LH2 antenna complexes of purple bacteria occur, depending on light conditions, in various different spectroscopic forms, with a similar structure but different absorption spectra. The differences are related to point changes in the primary amino acid sequence, but the molecular-level relationship between these changes and the resulting spectrum is still not well understood. We undertook a systematic quantum chemical analysis of all the main factors that contribute to the exciton structure, looking at how the environment modulates site energies and couplings in the B800-850 and B800-820 spectroscopic forms of LH2. A multiscale approach combining quantum chemistry and an atomistic classical embedding has been used where mutual polarization effects between the two parts are taken into account. We find that the loss of hydrogen bonds following amino acid changes can only explain a part of the observed blue-shift in the B850 band. The coupling of excitonic states to charge-transfer states, which is different in the two forms, contributes with a similar amount to the overall blue-shift.
Assuntos
Proteínas de Bactérias/fisiologia , Complexos de Proteínas Captadores de Luz/fisiologia , Rodopseudomonas/fisiologia , Proteínas de Bactérias/química , Domínio Catalítico , Complexos de Proteínas Captadores de Luz/química , Conformação ProteicaRESUMO
In higher plant thylakoids, the heterogeneous distribution of photosynthetic protein complexes is a determinant for the formation of grana, stacks of membrane discs that are densely populated with Photosystem II (PSII) and its light harvesting complex (LHCII). PSII associates with LHCII to form the PSII-LHCII supercomplex, a crucial component for solar energy conversion. Here, we report a biochemical, structural and functional characterization of pairs of PSII-LHCII supercomplexes, which were isolated under physiologically-relevant cation concentrations. Using single-particle cryo-electron microscopy, we determined the three-dimensional structure of paired C2S2M PSII-LHCII supercomplexes at 14 Å resolution. The two supercomplexes interact on their stromal sides through a specific overlap between apposing LHCII trimers and via physical connections that span the stromal gap, one of which is likely formed by interactions between the N-terminal loops of two Lhcb4 monomeric LHCII subunits. Fast chlorophyll fluorescence induction analysis showed that paired PSII-LHCII supercomplexes are energetically coupled. Molecular dynamics simulations revealed that additional flexible physical connections may form between the apposing LHCII trimers of paired PSII-LHCII supercomplexes in appressed thylakoid membranes. Our findings provide new insights into how interactions between pairs of PSII-LHCII supercomplexes can link adjacent thylakoids to mediate the stacking of grana membranes.
Assuntos
Clorofila/química , Complexos de Proteínas Captadores de Luz/ultraestrutura , Complexo de Proteína do Fotossistema II/ultraestrutura , Pisum sativum/química , Folhas de Planta/química , Tilacoides/química , Sítios de Ligação , Microscopia Crioeletrônica , Complexos de Proteínas Captadores de Luz/fisiologia , Simulação de Dinâmica Molecular , Pisum sativum/fisiologia , Complexo de Proteína do Fotossistema II/fisiologia , Folhas de Planta/fisiologia , Ligação Proteica , Conformação Proteica em alfa-Hélice , Conformação Proteica em Folha beta , Domínios e Motivos de Interação entre Proteínas , Tilacoides/ultraestruturaRESUMO
During the first steps of photosynthesis, the energy of impinging solar photons is transformed into electronic excitation energy of the light-harvesting biomolecular complexes. The subsequent energy transfer to the reaction center is commonly rationalized in terms of excitons moving on a grid of biomolecular chromophores on typical timescales [Formula: see text]100 fs. Today's understanding of the energy transfer includes the fact that the excitons are delocalized over a few neighboring sites, but the role of quantum coherence is considered as irrelevant for the transfer dynamics because it typically decays within a few tens of femtoseconds. This orthodox picture of incoherent energy transfer between clusters of a few pigments sharing delocalized excitons has been challenged by ultrafast optical spectroscopy experiments with the Fenna-Matthews-Olson protein, in which interference oscillatory signals up to 1.5 ps were reported and interpreted as direct evidence of exceptionally long-lived electronic quantum coherence. Here, we show that the optical 2D photon echo spectra of this complex at ambient temperature in aqueous solution do not provide evidence of any long-lived electronic quantum coherence, but confirm the orthodox view of rapidly decaying electronic quantum coherence on a timescale of 60 fs. Our results can be considered as generic and give no hint that electronic quantum coherence plays any biofunctional role in real photoactive biomolecular complexes. Because in this structurally well-defined protein the distances between bacteriochlorophylls are comparable to those of other light-harvesting complexes, we anticipate that this finding is general and directly applies to even larger photoactive biomolecular complexes.
Assuntos
Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/fisiologia , Transferência de Energia/fisiologia , Complexos de Proteínas Captadores de Luz/metabolismo , Complexos de Proteínas Captadores de Luz/fisiologia , Proteínas de Bactérias/química , Bacterioclorofilas/metabolismo , Complexos de Proteínas Captadores de Luz/química , Fótons , Fotossíntese/fisiologia , Teoria Quântica , Análise Espectral/métodosRESUMO
Bigelowiella natans is a mixotrophic flagellate and member of the chlorarachniophytes (Rhizaria), whose plastid is derived from a green algal endosymbiont. With the completion of the B. natans nuclear genome we are able to begin the analysis of the structure, function and evolution of the photosynthetic apparatus. B. natans has undergone substantial changes in photosystem structure during the evolution of the plastid from a green alga. While Photosystem II (PSII) composition is well conserved, Photosystem I (PSI) composition has undergone a dramatic reduction in accessory protein subunits. Coinciding with these changes, there was a loss of green algal LHCI orthologs while the PSII-like antenna system has the expected green algal-like proteins (encoded by genes Lhcbm1-8, Lhcb4). There are also a collection of LHCX-like proteins, which are commonly associated with stramenopiles and other eukaryotes with red-algal derived plastids, along with two other unique classes of LHCs- LHCY and LHCZ- whose function remains cryptic. To understand the regulation of the LHC gene family as an initial probe of function, we conducted an RNA-seq experiment under a short-term, high-light (HL) and low-light stress. The most abundant LHCII transcript (Lhcbm6) plus two other LHCBM types (Lhcbm1, 2) were down regulated under HL and up-regulated following a shift to very-low light (VL), as is common in antenna specializing in light harvesting. Many of the other LHCII and LHCY genes had a small, but significant increase in HL and most were only moderately affected under VL. The LHCX and LHCZ genes, however, had a strong up-regulation under HL-stress and most declined under VL, suggesting that they primarily have a role in photoprotection. This contrasts to the LHCY family that is only moderately responsive to light and a much higher basal level of expression, despite being within the LHCSR/LHCX clade. The expression of LHCX/Z proteins under HL-stress may be related to the induction of long-term, non-photochemical quenching mechanisms.
Assuntos
Complexos de Proteínas Captadores de Luz/metabolismo , Rhizaria/metabolismo , Aclimatação , Evolução Biológica , Fluorescência , Regulação da Expressão Gênica , Complexos de Proteínas Captadores de Luz/fisiologia , Estrutura Molecular , Complexo de Proteína do Fotossistema I/genética , Complexo de Proteína do Fotossistema I/metabolismo , Complexo de Proteína do Fotossistema II/genética , Complexo de Proteína do Fotossistema II/metabolismo , Filogenia , Rhizaria/genética , Rhizaria/fisiologia , Análise de Sequência de DNA , Homologia de SequênciaRESUMO
In bacterial photosynthesis reaction center-light-harvesting 1 (RC-LH1) complexes trap absorbed solar energy by generating a charge separated state. Subsequent electron and proton transfers form a quinol, destined to diffuse to the cytochrome bc1 complex. In bacteria such as Rhodobacter (Rba.) sphaeroides and Rba. capsulatus the PufX polypeptide creates a channel for quinone/quinol traffic across the LH1 complex that surrounds the RC, and it is therefore essential for photosynthetic growth. PufX also plays a key role in dimerization of the RC-LH1-PufX core complex, and the structure of the Rba. sphaeroides complex shows that the PufX C-terminus, particularly the region from X49-X53, likely mediates association of core monomers. To investigate this putative interaction we analysed mutations PufX R49L, PufX R53L, PufX R49/53L and PufX G52L by measuring photosynthetic growth, fractionation of detergent-solubilised membranes, formation of 2-D crystals and electron microscopy. We show that these mutations do not affect assembly of PufX within the core or photosynthetic growth but they do prevent dimerization, consistent with predictions from the RC-LH1-PufX structure. We obtained low resolution structures of monomeric core complexes with and without PufX, using electron microscopy of negatively stained single particles and 3D reconstruction; the monomeric complex with PufX corresponds to one half of the dimer structure whereas LH1 completely encloses the RC if the gene encoding PufX is deleted. On the basis of the insights gained from these mutagenesis and structural analyses we propose a sequence for assembly of the dimeric RC-LH1-PufX complex.