Peripheral Light-harvesting Antenna Research Articles

Cryo-electron microscopy structure of the intact photosynthetic light-harvesting antenna-reaction center complex from a green sulfur bacterium.

The photosynthetic reaction center complex (RCC) of green sulfur bacteria (GSB) consists of the membrane-imbedded RC core and the peripheric energy transmitting proteins called Fenna-Matthews-Olson (FMO). Functionally, FMO transfers the absorbed energy from a huge peripheral light-harvesting antenna named chlorosome to the RC core where charge separation occurs. In vivo, one RC was found to bind two FMOs, however, the intact structure of RCC as well as the energy transfer mechanism within RCC remain to be clarified. Here we report a structure of intact RCC which contains a RC core and two FMO trimers from a thermophilic green sulfur bacterium Chlorobaculum tepidum at 2.9 Å resolution by cryo-electron microscopy. The second FMO trimer is attached at the cytoplasmic side asymmetrically relative to the first FMO trimer reported previously. We also observed two new subunits (PscE and PscF) and the N-terminal transmembrane domain of a cytochrome-containing subunit (PscC) in the structure. These two novel subunits possibly function to facilitate the binding of FMOs to RC core and to stabilize the whole complex. A new bacteriochlorophyll (numbered as 816) was identified at the interspace between PscF and PscA-1, causing an asymmetrical energy transfer from the two FMO trimers to RC core. Based on the structure, we propose an energy transfer network within this photosynthetic apparatus.

Identification of the Optimal Light Harvesting Antenna Size for High-Light Stress Mitigation in Plants.

One of the major constraints limiting biomass production in autotrophs is the low thermodynamic efficiency of photosynthesis, ranging from 1 to 4%. Given the absorption spectrum of photosynthetic pigments and the spectral distribution of sunlight, photosynthetic efficiencies as high as 11% are possible. It is well-recognized that the greatest thermodynamic inefficiencies in photosynthesis are associated with light absorption and conversion of excited states into chemical energy. This is due to the fact that photosynthesis light saturates at one quarter full sunlight intensity in plants resulting in the dissipation of excess energy as heat, fluorescence and through the production of damaging reactive oxygen species. Recently, it has been demonstrated that it is possible to adjust the size of the light harvesting antenna over a broad range of optical cross sections through targeted reductions in chlorophyll b content, selectively resulting in reductions of the peripheral light harvesting antenna size, especially in the content of Lhcb3 and Lhcb6. We have examined the impact of alterations in light harvesting antenna size on the amplitude of photoprotective activity and the evolutionary fitness or seed production in Camelina grown at super-saturating and sub-saturating light intensities to gain an understanding of the driving forces that lead to the selection for light harvesting antenna sizes best fit for a range of light intensities. We demonstrate that plants having light harvesting antenna sizes engineered for the greatest photosynthetic efficiency also have the greatest capacity to mitigate high light stress through non-photochemical quenching and reduction of reactive oxygen associated damage. Under sub-saturating growth light intensities, we demonstrate that the optimal light harvesting antenna size for photosynthesis and seed production is larger than that for plants grown at super-saturating light intensities and is more similar to the antenna size of wild-type plants. These results suggest that the light harvesting antenna size of plants is designed to maximize fitness under low light conditions such as occurs in shaded environments and in light competition with other plants.

Stark fluorescence spectroscopy on peridinin-chlorophyll-protein complex of dinoflagellate, Amphidinium carterae.

Because of their peculiar but intriguing photophysical properties, peridinin-chlorophyll-protein complexes (PCPs), the peripheral light-harvesting antenna complexes of photosynthetic dinoflagellates have been unique targets of multidimensional theoretical and experimental investigations over the last few decades. The major light-harvesting chlorophyll a (Chl a) pigments of PCP are hypothesized to be spectroscopically heterogeneous. To study the spectral heterogeneity in terms of electrostatic parameters, we, in this study, implemented Stark fluorescence spectroscopy on PCP isolated from the dinoflagellate Amphidinium carterae. The comprehensive theoretical modeling of the Stark fluorescence spectrum with the help of the conventional Liptay formalism revealed the simultaneous presence of three emission bands in the fluorescence spectrum of PCP recorded upon excitation of peridinin. The three emission bands are found to possess different sets of electrostatic parameters with essentially increasing magnitude of charge-transfer character from the blue to redder ones. The different magnitudes of electrostatic parameters give good support to the earlier proposition that the spectral heterogeneity in PCP results from emissive Chl a clusters anchored at a different sites and domains within the protein network.

Carotenoid-to-bacteriochlorophyll energy transfer through vibronic coupling in LH2 from Phaeosprillum molischianum

The peripheral light-harvesting antenna complex (LH2) of purple photosynthetic bacteria is an ideal testing ground for models of structure–function relationships due to its well-determined molecular structure and ultrafast energy deactivation. It has been the target for numerous studies in both theory and ultrafast spectroscopy; nevertheless, certain aspects of the convoluted relaxation network of LH2 lack a satisfactory explanation by conventional theories. For example, the initial carotenoid-to-bacteriochlorophyll energy transfer step necessary on visible light excitation was long considered to follow the Förster mechanism, even though transfer times as short as 40 femtoseconds (fs) have been observed. Such transfer times are hard to accommodate by Förster theory, as the moderate coupling strengths found in LH2 suggest much slower transfer within this framework. In this study, we investigate LH2 from Phaeospirillum (Ph.) molischianum in two types of transient absorption experiments—with narrowband pump and white-light probe resulting in 100 fs time resolution, and with degenerate broadband 10 fs pump and probe pulses. With regard to the split Qx band in this system, we show that vibronically mediated transfer explains both the ultrafast carotenoid-to-B850 transfer, and the almost complete lack of transfer to B800. These results are beyond Förster theory, which predicts an almost equal partition between the two channels.

An Ab Initio Description of the Excitonic Properties of LH2 and Their Temperature Dependence.

The spectroscopic properties of light-harvesting (LH) antennae in photosyntehtic organisms represent a fingerprint that is unique for each specific pigment-protein complex. Because of that, spectroscopic observations are generally combined with structural data from X-ray crystallography to obtain an indirect representation of the excitonic properties of the system. Here, an alternative strategy is presented which goes beyond this empirical approach and introduces an ab initio computational description of both structural and electronic properties and their dependence on the temperature. The strategy is applied to the peripheral light-harvesting antenna complex (LH2) present in purple bacteria. By comparing this model with the one based on the crystal structure, a detailed, molecular level explanation of the absorption and circular dichroism (CD) spectra and their temperature dependence is achieved. The agreement obtained with the experiments at both low and room temperature lays the groundwork for an atomistic understanding of the excitation dynamics in the LH2 system.

Truncated light-harvesting chlorophyll antenna size in Chlorella vulgaris improves biomass productivity

Microalgae have been proposed as eco-friendly feedstocks for biodiesel production, because they accumulate large amounts of lipids and increase their biomass through photosynthesis. However, the photosynthetic efficiency of microalgae is too low for this strategy to be economically feasible. In an effort to overcome this issue, random mutants with reduced chlorophyll antenna size were generated by ethyl methanesulfonate (EMS)-mediated mutagenesis of Chlorella vulgaris. The antenna size mutant, herein designated E5, exhibited 56.5 and 75.8 % decreases in chlorophyll a and b contents, respectively, with significant reductions in the expression levels of peripheral light-harvesting antenna proteins in photosystem II. The saturated photosynthetic activity and electron transport rate of the E5 mutant were significantly higher and also showed reduced non-photochemical quenching (NPQ), compared to those of the wild type. Consequentially, the E5 mutant cultures achieved 44.5 % improvement in biomass productivity under high light (200 μmol photons m−2 s−1). These results suggest that improving the photosynthetic efficiency of microalgae could greatly enhance their biomass production, and such mutant strains can be applicable for large-scale outdoor cultivation which is typically exposed to high light intensity.

Clades of Photosynthetic Bacteria Belonging to the Genus Rhodopseudomonas Show Marked Diversity in Light-Harvesting Antenna Complex Gene Composition and Expression.

Many photosynthetic bacteria have peripheral light-harvesting (LH) antenna complexes that increase the efficiency of light energy capture. The purple nonsulfur photosynthetic bacterium Rhodopseudomonas palustris produces different types of LH complexes under high light intensities (LH2 complex) and low light intensities (LH3 and LH4 complexes). There are multiple pucBA operons that encode the α and β peptides that make up these complexes. However, low-resolution structures, amino acid similarities between the complexes, and a lack of transcription analysis have made it difficult to determine the contributions of different pucBA operons to the composition and function of different LH complexes. It was also unclear how much diversity of LH complexes exists in R.palustris and affiliated strains. To address this, we undertook an integrative genomics approach using 20 sequenced strains. Gene content analysis revealed that even closely related strains have differences in their pucBA gene content. Transcriptome analyses of the strains grown under high light and low light revealed that the patterns of expression of the pucBA operons varied among strains grown under the same conditions. We also found that one set of LH2 complex proteins compensated for the lack of an LH4 complex under low light intensities but not under extremely low light intensities, indicating that there is functional redundancy between some of the LH complexes under certain light intensities. The variation observed in LH gene composition and expression in Rhodopseudomonas strains likely reflects how they have evolved to adapt to light conditions in specific soil and water microenvironments. IMPORTANCERhodopseudomonas palustris is a phototrophic purple nonsulfur bacterium that adapts its photosystem to allow growth at a range of light intensities. It does this by adjusting the amount and composition of peripheral light-harvesting (LH) antenna complexes that it synthesizes. Rhodopseudomonas strains are notable for containing numerous sets of light-harvesting genes. We determined the diversity of LH complexes and their transcript levels during growth under high and low light intensities in 20 sequenced genomes of strains related to the species Rhodopseudomonas palustris. The data obtained are a resource for investigators with interests as wide-ranging as the biophysics of photosynthesis, the ecology of phototrophic bacteria, and the use of photosynthetic bacteria for biotechnology applications.

Organization of chlorophyll biosynthesis and insertion of chlorophyll into the chlorophyll-binding proteins in chloroplasts.

Oxygenic photosynthesis requires chlorophyll (Chl) for the absorption of light energy, and charge separation in the reaction center of photosystem I and II, to feed electrons into the photosynthetic electron transfer chain. Chl is bound to different Chl-binding proteins assembled in the core complexes of the two photosystems and their peripheral light-harvesting antenna complexes. The structure of the photosynthetic protein complexes has been elucidated, but mechanisms of their biogenesis are in most instances unknown. These processes involve not only the assembly of interacting proteins, but also the functional integration of pigments and other cofactors. As a precondition for the association of Chl with the Chl-binding proteins in both photosystems, the synthesis of the apoproteins is synchronized with Chl biosynthesis. This review aims to summarize the present knowledge on the posttranslational organization of Chl biosynthesis and current attempts to envision the proceedings of the successive synthesis and integration of Chl into Chl-binding proteins in the thylakoid membrane. Potential auxiliary factors, contributing to the control and organization of Chl biosynthesis and the association of Chl with the Chl-binding proteins during their integration into photosynthetic complexes, are discussed in this review.

Heat- and light-induced detachment of the light-harvesting antenna complexes of photosystem I in isolated stroma thylakoid membranes

The multisubunit pigment–protein complex of photosystem I (PSI) consists of a core and peripheral light-harvesting antenna (LHCI). PSI is thought to be a rather rigid system and very little is known about its structural and functional flexibility. Recent data, however, suggest LHCI detachment from the PSI supercomplex upon heat and light treatments. Furthermore, it was suggested that the splitting off of LHCI acts as a safety valve for PSI core upon photoinhibition (Alboresi et al., 2009).In this work we analyzed the heat- and light-induced reorganizations in isolated PSI vesicles (stroma membrane vesicles enriched in PSI). Using differential scanning calorimetry we revealed a stepwise disassembly of PSI supercomplex above 50°C. Circular dichroism, sucrose gradient centrifugation and 77K fluorescence experiments identified the sequence of events of PSI destabilization: 3min heating at 60°C or 40min white light illumination at 25°C resulted in pronounced Lhca1/4 detachment from the PSI supercomplex, which is then followed by the degradation of Lhca2/3.The similarity of the main structural effects due to heat and light treatments supports the notion that thermo-optic mechanism, structural changes induced by ultrafast local thermal transients, which has earlier been shown to be responsible for structural changes in the antenna system of photosystem II, can also regulate the assembly and functioning of PSI antenna.

Changes in the Energy Transfer Pathways within Photosystem II Antenna Induced by Xanthophyll Cycle Activity

Energy transfer pathways between photosystem II (PSII) antenna complexes in intact thylakoid membranes have been studied using low-temperature (77 K) excitation fluorescence spectroscopy. The focus of this study was to see whether de-epoxidation of violaxanthin into zeaxanthin causes any alterations in the energetic couplings between the core antenna complexes CP43 and CP47 and the peripheral light-harvesting antenna (LHCII). It was discovered that the appearance of zeaxanthin caused characteristic alterations in the PSII excitation fluorescence spectra in the Soret-band region. While in the dark violaxanthin was found to be largely uncoupled from any emitting chlorophylls, its intensive de-epoxidation resulted in the appearance of two additional bands at 509 and 492 nm. The former was attributed to weak coupling of zeaxanthin to emitters in the CP43 and LHCII complexes and the latter to efficient coupling of violaxanthin of the CP29 complex to emitters in the CP43, CP47, and LHCII complexes. The role of CP29-bound violaxanthin is discussed in light of both its efficient energetic coupling and strong physical binding to this complex. The finding that zeaxanthin is energetically coupled to chlorophyll a emitters of the PSII antenna is discussed with respect to its suggested role as a quencher involved in photoprotective energy dissipation, or non-photochemical quenching (NPQ), in the photosynthetic membrane.

A resonance mechanism of efficient energy transfer mediated by Fenna-Matthews-Olson complex

The Wigner-Weisskopf-type model developed by Alicki and Giraldi [J. Phys. B 44, 154020 (2011)] is applied to the biological process of energy transfer from a large peripheral light harvesting antenna to the reaction center. This process is mediated by the Fenna-Matthews-Olson (FMO) photosynthetic complex with a remarkably high efficiency. The proposed model provides a simple resonance mechanism of this phenomenon employing exciton coherent motion and is described by analytical formulas. A coupling to the vibrational environment is a necessary component of this mechanism as well as a fine-tuning of the FMO complex Hamiltonian. The role of the relatively strong coupling to the energy sink in achieving the resonance condition and the absence of heating of the vibrational environment are emphasized.

Polarization single complex imaging of circular photosynthetic antenna

Single complex fluorescence polarization spectroscopy is applied to study the peripheral light harvesting antenna (LH2) from photosynthetic purple bacterium Rhodopseudomonas (Rps.) acidophila. The measured two-dimensional excitation-emission polarization plots are used to construct geometric representation for the absorbing B800 and emitting B850 as ellipses. The shape and orientation of the ellipses is discussed in terms of tilted LH2 complexes where emission occurs from energetically disordered B850 excitons.

Temperature and Ionic Strength Effects on the Chlorosome Light-Harvesting Antenna Complex

Chlorosomes, the peripheral light-harvesting antenna complex from green photosynthetic bacteria, are the largest and one of the most efficient light-harvesting antenna complexes found in nature. In contrast to other light-harvesting antennas, chlorosomes are constructed from more than 150,000 self-assembled bacteriochlorophylls (BChls) and contain relatively few proteins that play secondary roles. These unique properties have led to chlorosomes as an attractive candidate for developing biohybrid solar cell devices. In this article, we investigate the temperature and ionic strength effects on the viability of chlorosomes from the photosynthetic green bacterium Chloroflexus aurantiacus using small-angle neutron scattering and dynamic light scattering. Our studies indicate that chlorosomes remain intact up to 75 °C and that salt induces the formation of large aggregates of chlorosomes. No internal structural changes are observed for the aggregates. The salt-induced aggregation, which is a reversible process, is more efficient with divalent metal ions than with monovalent metal ions. Moreover, with treatment at 98 °C for 2 min, the bulk of the chlorosome pigments are undamaged, while the baseplate is destroyed. Chlorosomes without the baseplate remain rodlike in shape and are 30-40% smaller than with the baseplate attached. Further, chlorosomes are stable from pH 5.5 to 11.0. Together, this is the first time such a range of characterization tools have been used for chlorosomes, and this has enabled elucidation of properties that are not only important to understanding their functionality but also may be useful in biohybrid devices for effective light harvesting.

ChemInform Abstract: Photoinduced Energy Transfer in Artificial Photosynthetic Systems

AbstractReview: 221 refs.

Hydrogen production by recombinant strains of Rhodobacter sphaeroides using a modified photosynthetic apparatus

Hydrogen release by recombinant strains of Rhodobacter sphaeroides pRK puf DD13 without a peripheral light-harvesting antenna complex and pRK puf deltaLM1 which is able to synthesize both antenna complexes, both of which were grown in conditions of nitrogen limitation, has been studied. The velocity of hydrogen release depended on light intensity. At high cell concentration (0.91 g l(-1)) of pRK puf DD 13, velocity was maximal at 2270 W m(-2) and was equal to 144.7 ml l(-1) h(-1) that evidences to an opportunity to increase the volume velocity of hydrogen release by application of the strains with low content of pigments.

The composition and structure of photosystem I-associated antenna from Cyanidioschyzon merolae

Red algae contain two types of light-harvesting antenna systems, the phycobilisomes and chlorophyll a binding polypeptides (termed Lhcr), which expand the light-harvesting capacity of the photosynthetic reaction centers. In this study, photosystem I (PSI) and its associated light-harvesting proteins were isolated from the red alga Cyanidioschyzon merolae. The structural and functional properties of the largest PSI particles observed were investigated by biochemical characterization, mass spectrometry, fluorescence emission and excitation spectroscopy, and transmission electron microscopy. Our data provide strong evidence for a stable PSI complex in red algae that possesses two distinct types of functional peripheral light-harvesting antenna complex, comprising both Lhcr and a PSI-linked phycobilisome sub-complex. We conclude that the PSI antennae system of red algae represents an evolutionary intermediate between the prokaryotic cyanobacteria and other eukaryotes, such as green algae and vascular plants.

Theoretical examination of quantum coherence in a photosynthetic system at physiological temperature

The observation of long-lived electronic coherence in a photosynthetic pigment-protein complex, the Fenna-Matthews-Olson (FMO) complex, is suggestive that quantum coherence might play a significant role in achieving the remarkable efficiency of photosynthetic electronic energy transfer (EET), although the data were acquired at cryogenic temperature [Engel GS, et al. (2007) Evidence for wavelike energy transfer through quantum coherence in photosynthetic systems. Nature 446:782-786]. In this paper, the spatial and temporal dynamics of EET through the FMO complex at physiological temperature are investigated theoretically. The numerical results reveal that quantum wave-like motion persists for several hundred femtoseconds even at physiological temperature, and suggest that the FMO complex may work as a rectifier for unidirectional energy flow from the peripheral light-harvesting antenna to the reaction center complex by taking advantage of quantum coherence and the energy landscape of pigments tuned by the protein scaffold. A potential role of quantum coherence is to overcome local energetic traps and aid efficient trapping of electronic energy by the pigments facing the reaction center complex.

Evidence for wavelike energy transfer through quantum coherence in photosynthetic systems

Photosynthetic complexes are exquisitely tuned to capture solar light efficiently, and then transmit the excitation energy to reaction centres, where long term energy storage is initiated. The energy transfer mechanism is often described by semiclassical models that invoke 'hopping' of excited-state populations along discrete energy levels. Two-dimensional Fourier transform electronic spectroscopy has mapped these energy levels and their coupling in the Fenna-Matthews-Olson (FMO) bacteriochlorophyll complex, which is found in green sulphur bacteria and acts as an energy 'wire' connecting a large peripheral light-harvesting antenna, the chlorosome, to the reaction centre. The spectroscopic data clearly document the dependence of the dominant energy transport pathways on the spatial properties of the excited-state wavefunctions of the whole bacteriochlorophyll complex. But the intricate dynamics of quantum coherence, which has no classical analogue, was largely neglected in the analyses-even though electronic energy transfer involving oscillatory populations of donors and acceptors was first discussed more than 70 years ago, and electronic quantum beats arising from quantum coherence in photosynthetic complexes have been predicted and indirectly observed. Here we extend previous two-dimensional electronic spectroscopy investigations of the FMO bacteriochlorophyll complex, and obtain direct evidence for remarkably long-lived electronic quantum coherence playing an important part in energy transfer processes within this system. The quantum coherence manifests itself in characteristic, directly observable quantum beating signals among the excitons within the Chlorobium tepidum FMO complex at 77 K. This wavelike characteristic of the energy transfer within the photosynthetic complex can explain its extreme efficiency, in that it allows the complexes to sample vast areas of phase space to find the most efficient path.

The study of photo-induced ultrafast dynamics in light-harvesting complex LH2 of purple bacteria

In this paper, we introduce the photo-induced ultrafast dynamics taking place in the peripheral light harvesting antenna LH2 from purple bacteria Rhodobacter sphaeroides by using absorption, fluorescence emission and ultrafast spectroscopic techniques. Three kinds of LH2 samples, pH treated LH2 (complete removal of B800 pigments), carotenoid mutated LH2 (GM 309) and electrochemical oxidation treated LH2 were used in comparison with native LH2 to investigate the mechanism of photo-induced ultrafast energy transfer within the LH2 complex.

A Chlorophyll a/b-binding Protein Homolog That Is Induced by Iron Deficiency Is Associated with Enlarged Photosystem I Units in the Eucaryotic Alga Dunaliella salina

Adaptation of the halotolerant alga Dunaliella salina to iron deprivation involves extensive changes of chloroplast morphology, photosynthetic activities, and induction of a major 45-kDa chloroplast protein termed Tidi. Partial amino acid sequencing of proteolytic peptides suggested that Tidi resembles chlorophyll a/b-binding proteins which compose light-harvesting antenna complexes (LHC) (Varsano, T., Kaftan, D., and Pick, U. (2003) J. Plant Nutr. 26, 2197-2210). Here we show that Tidi shares the highest amino acid sequence similarity with light-harvesting I chlorophyll a/b-binding proteins from higher plants but has an extended proline-rich N-terminal domain. The accumulation of Tidi is reversed by iron supplementation, and its level is inversely correlated with photosystem I (PS-I) reaction center proteins. In native gel electrophoresis, Tidi co-migrates with enlarged PS-I-LHC-I super-complexes. Single particle electron microscopy analysis revealed that PS-I units from iron-deficient cells are larger (31 and 37 nm in diameter) than PS-I units from control cells (22 nm). The 77 K chlorophyll fluorescence emission spectra of isolated complexes suggest that the Tidi-LHC-I antenna are functionally coupled to the reaction centers of PS-I. These findings indicate that Tidi acts as an accessory antenna of PS-I. The enlargement of PS-I antenna in algae and in cyanobacteria under iron deprivation suggests a common limitation that requires rebalancing of the energy distribution between the two photosystems.