Showing papers on "Photosynthetic reaction centre published in 2023"

Rhodobacter capsulatus forms a compact crescent-shaped LH1–RC photocomplex

Abstract: Abstract Rhodobacter ( Rba .) capsulatus has been a favored model for studies of all aspects of bacterial photosynthesis. This purple phototroph contains PufX, a polypeptide crucial for dimerization of the light-harvesting 1–reaction center (LH1–RC) complex, but lacks protein-U, a U-shaped polypeptide in the LH1–RC of its close relative Rba. sphaeroides . Here we present a cryo-EM structure of the Rba. capsulatus LH1–RC purified by DEAE chromatography. The crescent-shaped LH1–RC exhibits a compact structure containing only 10 LH1 αβ-subunits. Four αβ-subunits corresponding to those adjacent to protein-U in Rba. sphaeroides were absent. PufX in Rba. capsulatus exhibits a unique conformation in its N-terminus that self-associates with amino acids in its own transmembrane domain and interacts with nearby polypeptides, preventing it from interacting with proteins in other complexes and forming dimeric structures. These features are discussed in relation to the minimal requirements for the formation of LH1–RC monomers and dimers, the spectroscopic behavior of both the LH1 and RC, and the bioenergetics of energy transfer from LH1 to the RC.

Advances in the Spectroscopic and Structural Characterization of Core Light-Harvesting Complexes from Purple Phototrophic Bacteria.

Abstract: Purple phototrophic bacteria are ancient anoxygenic phototrophs and attractive research tools because they capture light energy in the near-infrared (NIR) region of the spectrum and transform it into chemical energy by way of uphill energy transfers. The heart of this reaction occurs in light-harvesting 1-reaction center (LH1-RC) complexes, which are the simplest model systems for understanding basic photosynthetic reactions within type-II (quinone-utilizing) reaction centers. In this Perspective, we highlight structure-function relationships concerning unresolved fundamental processes in purple bacterial photosynthesis, including the diversified light-harvesting capacity of LH1-associated BChl molecules, energies necessary for photoelectric conversion in the RC special pairs, and quinone transport mechanisms. Based on recent progress in the spectroscopic and structural analysis of LH1-RC complexes from a variety of purple phototrophs, we discuss several key factors for understanding how purple bacteria resource light energy in the inherently energy-poor NIR region of the electromagnetic spectrum.

Study on Heat Resistance of Peony Using Photosynthetic Indexes and Rapid Fluorescence Kinetics

Abstract: To investigate the effects of high-temperature stress on the chlorophyll fluorescence induction kinetics of peony and to determine indicators for the rapid screening of varieties responding to high temperatures, three four-year-old peony variety, ‘Fengdanbai’, ‘Huhong’, and ‘Yinhongqiaodui’, were selected as materials. The photosynthetic curves (Pn-PAR) and fast chlorophyll fluorescence curves (OJIP curves) of peony leaves were measured at different times under high-temperature stress conditions (40 °C), the changes in the photosynthetic characteristics of different peony varieties under high-temperature stress were analyzed, and the heat tolerance of peony was evaluated. The results showed that ‘Huhong’ grew well within 16 days, while all of the other varieties showed obvious wilting at 6–9 days. High temperatures damaged the structure and function of the photosystem of peony leaves, indicating that the maximum net photosynthetic rate (Pnmax), apparent quantum efficiency (AQE), maximum photochemical efficiency (Fv/Fm), and photosynthetic performance index (PIABS) all tended to decrease under high-temperature stress, while the rate of closing the PS II reaction center (Mo) and the absorption per reaction center (ABS/RC), the capture (TRo/RC), and the dissipation (Dio/RC) of light energy per reaction center showed an overall increasing trend. The ability to cope with high-temperature stress differed among varieties, and the heat tolerance was determined to be in the descending order of ‘Fengdanbai’ < ‘Yinhongqiaodui’ < ‘Huhong’. The correlation analysis among the parameters and the analysis of the morphological change patterns in peony leaves concluded that PIABS, Dio/RC, and Mo could be used as indicators of peony tolerance to high-temperature stress. The results of the study can provide a basis for the screening of heat-tolerant peony species and peony heat defense in the Jiangnan area.

Cryo-EM structure of the whole photosynthetic reaction center apparatus from the green sulfur bacterium Chlorobaculum tepidum.

Abstract: Light energy absorption and transfer are very important processes in photosynthesis. In green sulfur bacteria light is absorbed primarily by the chlorosomes and its energy is transferred via the Fenna-Matthews-Olson (FMO) proteins to a homodimeric reaction center (RC). Here, we report the cryogenic electron microscopic structure of the intact FMO-RC apparatus from Chlorobaculum tepidum at 2.5 Å resolution. The FMO-RC apparatus presents an asymmetric architecture and contains two FMO trimers that show different interaction patterns with the RC core. Furthermore, the two permanently bound transmembrane subunits PscC, which donate electrons to the special pair, interact only with the two large PscA subunits. This structure fills an important gap in our understanding of the transfer of energy from antenna to the electron transport chain of this RC and the transfer of electrons from reduced sulfur compounds to the special pair.

An exploratory steady-state redox model of photosynthetic linear electron transport for use in complete modeling of photosynthesis for broad applications.

Abstract: A photochemical model of photosynthetic electron transport (PET) is needed to integrate photophysics, photochemistry, and biochemistry to determine redox conditions of electron carriers and enzymes for plant stress assessment and mechanistically link sun-induced chlorophyll fluorescence to carbon assimilation for remotely sensing photosynthesis. Towards this goal, we derived photochemical equations governing the states and redox reactions of complexes and electron carriers along the PET chain. These equations allow the redox conditions of the mobile plastoquinone pool and the cytochrome b6 f complex (Cyt) to be inferred with typical fluorometry. The equations agreed well with fluorometry measurements from diverse C3 /C4 species across environments in the relationship between the PET rate and fraction of open photosystem II reaction centers. We found the oxidation of plastoquinol by Cyt is the bottleneck of PET, and genetically improving the oxidation of plastoquinol by Cyt may enhance the efficiency of PET and photosynthesis across species. Redox reactions and photochemical and biochemical interactions are highly redundant in their complex controls of PET. Although individual reaction rate constants cannot be resolved, they appear in parameter groups which can be collectively inferred with fluorometry measurements for broad applications. The new photochemical model developed enables advances in different fronts of photosynthesis research. This article is protected by copyright. All rights reserved.

Redox Potentials of Iron-Sulfur Clusters in Type I Photosynthetic Reaction Centers.

Abstract: The electron transfer pathways in type I photosynthetic reaction centers, such as photosystem I (PSI) and reaction centers from green sulfur bacteria (GsbRC), are terminated by two Fe4S4 clusters, FA and FB. The protein structures are the basis of understanding how the protein electrostatic environment interacts with the Fe4S4 clusters and facilitates electron transfer. Using the protein structures, we calculated the redox potential (Em) values for FA and FB in PSI and GsbRC, solving the linear Poisson-Boltzmann equation. The FA-to-FB electron transfer is energetically downhill in the cyanobacterial PSI structure, while it is isoenergetic in the plant PSI structure. The discrepancy arises from differences in the electrostatic influences of conserved residues, including PsaC-Lys51 and PsaC-Arg52, located near FA. The FA-to-FB electron transfer is slightly downhill in the GsbRC structure. Em(FA) and Em(FB) exhibit similar levels upon isolation of the membrane-extrinsic PsaC and PscB subunits from the PSI and GsbRC reaction centers, respectively. The binding of the membrane-extrinsic subunit at the heterodimeric/homodimeric reaction center plays a key role in tuning Em(FA) and Em(FB).

Properties and Crystal Structure of the Cereibacter sphaeroides Photosynthetic Reaction Center with Double Amino Acid Substitution I(L177)H + F(M197)H

Abstract: The photosynthetic reaction center of the purple bacterium Cereibacter sphaeroides with two site-directed mutations Ile-L177–His and M197 Phe–His is of double interest. The substitution I(L177)H results in strong binding of a bacteriochlorophyll molecule with L-subunit. The second mutation F(M197)H introduces a new H-bond between the C2-acetyl carbonyl group of the bacteriochlorophyll PB and His-M197, which is known to enhance the stability of the complex. Due to this H-bond, π -electron system of P finds itself connected to an extensive H-bonding network on the periplasmic surface of the complex. The crystal structure of the double mutant reaction center obtained with 2.6 Å resolution allows clarifying consequences of the Ile L177 – His substitution. The value of the P/P+ midpoint potential in the double mutant RC was found to be ~20 mV less than the sum of potentials measured in the two RCs with single mutations I(L177)H and F(M197)H. The protein environment of the BChls PA and BB were found to be similar to that in the RC with single substitution I(L177)H, whereas an altered pattern of the H-bonding networks was found in the vicinity of bacteriochlorophyll PB. The data obtained are consistent with our previous assumption on a correlation between the bulk of the H-bonding network connected with the π-electron system of the primary electron donor P and the value of its oxidation potential.

Absence of electron transfer-associated changes in the time-dependent X-ray free-electron laser structures of the photosynthetic reaction center

Abstract: Using the X-ray free-electron laser (XFEL) structures of the photosynthetic reaction center from Blastochloris viridis that show light-induced time-dependent structural changes [Dods, R.et al. (2021) Nature 589, 310-314], we investigated time-dependent changes in the energetics of the electron transfer pathway, considering the entire protein environment of the protein structures and titrating the redox active sites in the presence of all fully equilibrated titratable residues. In the dark and charge-separation intermediate structures, the calculated redox potential (Em) values for the accessory bacteriochlorophyll and bacteriopheophytin in the electron-transfer active branch (BL and HL) are higher than those in the electron-transfer inactive branch (BM and HM). However, the stabilization of the [PLPM]•+HL•– state owing to protein reorganization is not clearly observed in the Em(HL) values in the charge-separated 5-ps ([PLPM]•+HL•– state) structure. Furthermore, the expected chlorin ring deformation upon formation of HL•– (saddling mode) is absent in the HL geometry of the original 5-ps structure. These findings suggest that there is no clear link between the time-dependent structural changes and the electron transfer events in the XFEL structures.

Mapping Charge-Transfer Excitations in Bacteriochlorophyll Dimers from First Principles

Abstract: Photoinduced charge-transfer excitations are key to understand the primary processes of natural photosynthesis and for designing photovoltaic and photocatalytic devices. In this paper, we use Bacteriochlorophyll dimers extracted from the light harvesting apparatus and reaction center of a photosynthetic purple bacterium as model systems to study such excitations using first-principles numerical simulation methods. We distinguish four different regimes of intermolecular coupling, ranging from very weakly coupled to strongly coupled, and identify the factors that determine the energy and character of charge-transfer excitations in each case. We also construct an artificial dimer to systematically study the effects of intermolecular distance and orientation on charge-transfer excitations, as well as the impact of molecular vibrations on these excitations. Our results provide design rules for tailoring charge-transfer excitations in Bacteriochloropylls and related photoactive molecules, and highlight the importance of including charge-transfer excitations in accurate models of the excited-state structure and dynamics of Bacteriochlorophyll aggregates.

Understanding Primary Charge Separation in the Heliobacterial Reaction Center.

Abstract: The homodimeric reaction center of heliobacteria retains features of the ancestral reaction center and can thus provide insights into the evolution of photosynthesis. Primary charge separation is expected to proceed in a two-step mechanism along either of the two reaction center branches. We reveal the first charge-separation step from first-principles calculations based on time-dependent density functional theory with an optimally tuned range-separated hybrid and ab initio Born-Oppenheimer molecular dynamics: the electron is most likely localized on the electron transfer cofactor 3 (EC3, OH-chlorophyll a), and the hole on the adjacent EC2. Including substantial parts of the surrounding protein environment into the calculations shows that a distinct structural mechanism is decisive for the relative energetic positioning of the electronic excitations: specific charged amino acids in the vicinity of EC3 lower the energy of charge-transfer excitations and thus facilitate efficient charge separation. These results are discussed considering recent experimental insights.

Carotenoid assembly regulates quinone diffusion and the Roseiflexus castenholzii reaction center-light harvesting complex architecture

Abstract: Carotenoid (Car) pigments perform central roles in photosynthesis-related light harvesting (LH), photoprotection, and assembly of functional pigment-protein complexes. However, the relationships between Car depletion in the LH, assembly of the prokaryotic reaction center (RC)-LH complex, and quinone exchange are not fully understood. Here, we analyzed native RC-LH (nRC-LH) and Car-depleted RC-LH (dRC-LH) complexes in Roseiflexus castenholzii, a chlorosome-less filamentous anoxygenic phototroph that forms the deepest branch of photosynthetic bacteria. Newly identified exterior Cars functioned with the bacteriochlorophyll B800 to block the proposed quinone channel between LHαβ subunits in the nRC-LH, forming a sealed LH ring that was disrupted by transmembrane helices from cytochrome c and subunit X to allow quinone shuttling. dRC-LH lacked subunit X, leading to an exposed LH ring with a larger opening, which together accelerated the quinone exchange rate. We also assigned amino acid sequences of subunit X and two hypothetical proteins Y and Z that functioned in forming the quinone channel and stabilizing the RC-LH interactions. This study reveals the structural basis by which Cars assembly regulates the architecture and quinone exchange of bacterial RC-LH complexes. These findings mark an important step forward in understanding the evolution and diversity of prokaryotic photosynthetic apparatus.

Barley Cultivar Sarab 1 Has a Characteristic Region on the Thylakoid Membrane That Protects Photosystem I under Iron-Deficient Conditions

Abstract: The barley cultivar Sarab 1 (SRB1) can continue photosynthesis despite its low Fe acquisition potential via roots and dramatically reduced amounts of photosystem I (PSI) reaction-center proteins under Fe-deficient conditions. We compared the characteristics of photosynthetic electron transfer (ET), thylakoid ultrastructure, and Fe and protein distribution on thylakoid membranes among barley cultivars. The Fe-deficient SRB1 had a large proportion of functional PSI proteins by avoiding P700 over-reduction. An analysis of the thylakoid ultrastructure clarified that SRB1 had a larger proportion of non-appressed thylakoid membranes than those in another Fe-tolerant cultivar, Ehimehadaka-1 (EHM1). Separating thylakoids by differential centrifugation further revealed that the Fe-deficient SRB1 had increased amounts of low/light-density thylakoids with increased Fe and light-harvesting complex II (LHCII) than did EHM1. LHCII with uncommon localization probably prevents excessive ET from PSII leading to elevated NPQ and lower PSI photodamage in SRB1 than in EHM1, as supported by increased Y(NPQ) and Y(ND) in the Fe-deficient SRB1. Unlike this strategy, EHM1 may preferentially supply Fe cofactors to PSI, thereby exploiting more surplus reaction center proteins than SRB1 under Fe-deficient conditions. In summary, SRB1 and EHM1 support PSI through different mechanisms during Fe deficiency, suggesting that barley species have multiple strategies for acclimating photosynthetic apparatus to Fe deficiency.

Characterization of Tryptophan Oxidation Affecting D1 Degradation by FtsH in the Photosystem II Quality Control of Chloroplasts

Abstract: Light reaction of photosynthesis is one of the most important reactions for sustaining our environment. Photosystem II (PSII) is the initial site of photosynthetic electron transfer by water oxidation. Light in excess, however, causes the simultaneous production of singlet oxygen, a potent reactive oxygen species (ROS), leading to photo-oxidative damage in PSII. To maintain photosynthetic activity, the PSII reaction center protein D1, which is the primary target of unavoidable photo-oxidative damage, is efficiently degraded by FtsH protease. In PSII subunits, photo-oxidative modifications of several amino acids such as Trp have been indeed documented, whereas the linkage between such modifications and D1 degradation remains elusive. Here, we show that an oxidative post-translational modification of Trp residue at the N-terminal tail of D1 is correlated with D1 degradation by FtsH during high-light stress. We revealed that Arabidopsis mutant lacking FtsH2 had increased levels of oxidative Trp residues in D1, among which an N-terminal Trp-14 was distinctively localized in the stromal side. Further characterization of Trp-14 using chloroplast transformation in Chlamydomonas indicated that substitution of D1 Trp-14 to Phe, mimicking Trp oxidation enhanced FtsH-mediated D1 degradation under high light, although the substitution did not affect protein stability and PSII activity. Molecular dynamics simulation of PSII implies that both Trp-14 oxidation and Phe substitution cause fluctuation of D1 N-terminal tail. Furthermore, Trp-14 to Phe modification appeared to have an additive effect in the interaction between FtsH and PSII core in vivo. Together, our results suggest that the Trp oxidation at its N-terminus of D1 may be one of the key oxidations in the PSII repair, leading to processive degradation by FtsH. Significance Statement In photosynthetic organisms, maintenance of photosynthetic light reaction is manifested by so called Photosystem II (PSII) repair system, where the reaction center protein D1 is targeted to photo-oxidative damage and rapidly degraded by the processive protease FtsH. While this system is well known to cope with photoinhibition, the actual oxidation within the D1 polypeptide and its association to degradation remained elusive. Here, we characterized oxidative modification of tryptophan (Trp) residues in the PSII core, and hypothesize that the oxidation of N-terminal Trp is one of the key oxidations in the PSII repair, likely enhancing D1’s accessibility to FtsH.

Conversion of light into electricity in a semi-synthetic system based on photosynthetic bacterial chromatophores.

Abstract: Chromatophores (Chr) from photosynthetic nonsulfur purple bacterium Rhodobacter sphaeroides immobilized onto a Millipore membrane filter (MF) and sandwiched between two semiconductor indium tin oxide (ITO) electrodes (termed ITO|Chr - MF|ITO) have been used to measure voltage (ΔV) induced by continuous illumination. The maximum ΔV was detected in the presence of ascorbate / N,N,N'N'-tetramethyl-p-phenylenediamine couple, coenzyme UQ0, disaccaride trehalose and antimycin A, an inhibitor of cytochrome bc1 complex. In doing so, the light-induced electron transfer in the reaction centers was the major source of photovoltages. The stability of the voltage signal upon prolonged irradiation (>1 h) may be due to the maintenance of a conformation that is optimal for the functioning of integral protein complexes and stabilization of lipid bilayer membranes in the presence of trehalose. Retaining ∼70 % of the original photovoltage performance on the 30th day of storage at 23 °C in the dark under air was achieved after re-injection of fresh buffer (∼40 μL) containing redox mediators into the ITO|Chr - MF|ITO system. The approach we use is easy and can be extended to other biological intact systems (cells, thylakoid membranes) capable of converting energy of light.

De novo design of energy transfer proteins housing excitonically coupled chlorophyll special pairs

Abstract: Natural photosystems couple light harvesting to charge separation using a “special pair” of chlorophyll molecules that accepts excitation energy from the antenna and initiates an electron-transfer cascade. To investigate the photophysics of special pairs independent of complexities of native photosynthetic proteins, and as a first step towards synthetic photosystems for new energy conversion technologies, we designed C2-symmetric proteins that precisely position chlorophyll dimers. X-ray crystallography shows that one designed protein binds two chlorophylls in a binding orientation matching native special pairs, while a second positions them in a previously unseen geometry. Spectroscopy reveals excitonic coupling, and fluorescence lifetime imaging demonstrates energy transfer. We designed special pair proteins to assemble into 24-chlorophyll octahedral nanocages; the design model and cryo-EM structure are nearly identical. The design accuracy and energy transfer function of these special pair proteins suggest that de novo design of artificial photosynthetic systems is within reach of current computational methods.

Stabilization of Cereibacter sphaeroides Photosynthetic Reaction Center by the Introduction of Disulfide Bonds

Abstract: The photosynthetic reaction center of the purple nonsulfur bacterium Cereibacter sphaeroides is a useful model for the study of mechanisms of photoinduced electron transfer and a promising component for photo-bio-electrocatalytic systems. The basic research and technological applications of this membrane pigment-protein complex require effective approaches to increase its structural stability. In this work, a rational design approach to genetically modify the reaction centers by introducing disulfide bonds is used. This resulted in significantly increasing the thermal stability of some of the mutant pigment-protein complexes. The formation of the S-S bonds was confirmed by X-ray crystallography as well as SDS-PAGE, and the optical properties of the reaction centers were studied. The genetically modified reaction centers presented here preserved their ability for photochemical charge separation and could be of interest for basic science and biotechnology.

Triplet Delocalization over the Reaction Center Chlorophylls in Photosystem II.

Abstract: The triplet state of chlorophyll formed by charge recombination in photosystem II (PSII) is a precursor of harmful singlet oxygen. Although main localization of the triplet state on the monomeric chlorophyll, ChlD1, at cryogenic temperatures has been suggested, how the triplet state is delocalized on other chlorophylls remains unclear. Here, we investigated the distribution of the triplet state of chlorophyll in PSII using light-induced Fourier transform infrared (FTIR) difference spectroscopy. Measurements of triplet-minus-singlet FTIR difference spectra with PSII core complexes from cyanobacterial mutants, D1-V157H, D2-V156H, D2-H197A, and D1-H198A, in which the interactions of the 131-keto C═O groups of the reaction center chlorophylls, PD1, PD2, ChlD1, and ChlD2, respectively, were perturbed, identified the 131-keto C═O bands of the individual chlorophylls and showed that the triplet state is delocalized over all of these chlorophylls. It is suggested that the triplet delocalization plays important roles in the photoprotection and photodamage mechanisms in PSII.

Mapping charge-transfer excitations in Bacteriochlorophyll dimers from first principles

Abstract: Photoinduced charge-transfer excitations are key to understand the primary processes of natural photosynthesis and for designing photovoltaic and photocatalytic devices. In this paper, we use Bacteriochlorophyll dimers extracted from the light harvesting apparatus and reaction center of a photosynthetic purple bacterium as model systems to study such excitations using first-principles numerical simulation methods. We distinguish four different regimes of intermolecular coupling, ranging from very weakly coupled to strongly coupled, and identify the factors that determine the energy and character of charge-transfer excitations in each case. We also construct an artificial dimer to systematically study the effects of intermolecular distance and orientation on charge-transfer excitations, as well as the impact of molecular vibrations on these excitations. Our results provide design rules for tailoring charge-transfer excitations in Bacteriochloropylls and related photoactive molecules, and highlight the importance of including charge-transfer excitations in accurate models of the excited-state structure and dynamics of Bacteriochlorophyll aggregates.

The structure and assembly of reaction centre-light-harvesting 1 complexes in photosynthetic bacteria

Abstract: Abstract Chlorophototrophic organisms have a charge-separating reaction centre (RC) complex that receives energy from a dedicated light-harvesting (LH) antenna. In the purple phototrophic bacteria, these two functions are embodied by the ‘core’ photosynthetic component, the RC-LH1 complex. RC-LH1 complexes sit within a membrane bilayer, with the central RC wholly or partly surrounded by a curved array of LH1 subunits that bind a series of bacteriochlorophyll (BChl) and carotenoid pigments. Decades of research have shown that the absorption of light initiates a cascade of energy, electron, and proton transfers that culminate in the formation of a quinol, which is subsequently oxidized by the cytochrome bc1 complex. However, a full understanding of all these processes, from femtosecond absorption of light to millisecond quinone diffusion, requires a level of molecular detail that was lacking until the remarkable recent upsurge in the availability of RC-LH1 structures. Here, we survey 13 recently determined RC-LH1 assemblies, and we compare the precise molecular arrangements of pigments and proteins that allow efficient light absorption and the transfer of energy, electrons and protons. We highlight shared structural features, as well as differences that span the bound pigments and cofactors, the structures of individual subunits, the overall architecture of the complexes, and the roles of additional subunits newly identified in just one or a few species. We discuss RC-LH1 structures in the context of prior biochemical and spectroscopic investigations, which together enhance our understanding of the molecular mechanisms of photosynthesis in the purple phototrophic bacteria. A particular emphasis is placed on how the remarkable and unexpected structural diversity in RC-LH1 complexes demonstrates different evolutionary solutions for maximising pigment density for optimised light harvesting, whilst balancing the requirement for efficient quinone diffusion between RC and cytochrome bc1 complexes through the encircling LH1 complex.

Roadmap of electrons from donor side to the reaction center dimer of photosynthetic purple bacteria with mutated cytochromes

Abstract: Abstract In photosynthetic bacteria, the absorbed light drives the canonical cyclic electron transfer between the reaction center and the cytochrome bc 1 complexes via the pools of mobile electron carriers. If kinetic or structural barriers hinder the participation of the bc 1 complex in the cyclic flow of electrons, then the pools of mobile redox agents must supply the electrons for the multiple turnovers of the reaction center. These conditions were achieved by continuous high light excitation of intact cells of bacterial strains Rba. sphaeroides and Rvx. gelatinosus with depleted donor side cytochromes c 2 ( cycA ) and tetraheme cytochrome subunit ( pufC ), respectively. The graduate oxidation by ferricyanide reduced further the availability of electron donors of pufC . The electron transfer through the reaction center was tracked by absorption change of the dimer and by induction and relaxation of the bacteriochlorophyll fluorescence. The rate constants of the electron transfer (~ 3·10 3 s ‒1 ) from the mobile donors of Rvx. gelatinosus bound either to the RC ( pufC ) or to the tetraheme subunit (wild type) were similar. The electrons transferred through the reaction center dimer were supplied entirely by the donor pool, their number amounted about 5 in wild type Rvx. gelatinosus and decreased to 1 by exhaustion of the pool in pufC oxidized by ferricyanide. The complex shape of the measured function of the yield of fluorescence versus oxidized dimer revealed the contribution of two competing processes: the migration of the excitation energy among the photosynthetic units and the availability of electron donors to the oxidized dimer. The experimental results were simulated and rationalized by a simple kinetic model of the two-electron cycling of the acceptor side combined with aperiodic one-electron redox function of the donor side.

Photosynthesis in the near infrared: the γ subunit of <i>Blastochloris viridis</i> LH1 red-shifts absorption beyond 1000 nm

Abstract: The reaction centre (RC) in purple phototrophic bacteria is encircled by the primary light-harvesting complex 1 (LH1) antenna, forming the RC–LH1 ‘core’ complex. The Qy absorption maximum of LH1 complexes ranges from ∼875–960 nm in bacteriochlorophyll (BChl) a-utilising organisms, to 1018 nm in the BChl b-containing complex from Blastochloris (Blc.) viridis. The red-shifted absorption of the Blc. viridis LH1 was predicted to be due in part to the presence of the γ subunit unique to Blastochloris spp., which binds to the exterior of the complex and is proposed to increase packing and excitonic coupling of the BChl pigments. The study by Namoon et al. provides experimental evidence for the red-shifting role of the γ subunit and an evolutionary rationale for its incorporation into LH1. The authors show that cells producing RC–LH1 lacking the γ subunit absorb maximally at 972 nm, 46 nm to the blue of the wild-type organism. Wavelengths in the 900–1000 nm region of the solar spectrum transmit poorly through water, thus γ shifts absorption of LH1 to a region where photons have lower energy but are more abundant. Complementation of the mutant with a divergent copy of LH1γ resulted in an intermediate red shift, revealing the possibility of tuning LH1 absorption using engineered variants of this subunit. These findings provide new insights into photosynthesis in the lowest energy phototrophs and how the absorption properties of light-harvesting complexes are modified by the recruitment of additional subunits.

Ultrafast Energy Transfer Dynamics in a Cyanobacterial Light-Harvesting Phycobilisome

Abstract: The phycobilisomes (PBSs) of cyanobacteria and red algae are their primary light-harvesting antennas, which play key role in light harvesting and energy transportation to the photosynthetic reaction center with extraordinarily high efficiency. The mechanism of energy transfer in PBS should be investigated with a tight combination between biological structural information and an ultrafast time-resolved dynamic analysis. We recently demonstrated the study of energy transfer in PBSs from a thermophilic cyanobacterium, Thermosynechococcus vulcanus NIES 2134 (T. 2134), with the cryo-EM model resolved at a near-atomic resolution. The time-resolved fluorescence spectroscopy of the PBS with a sub-picosecond resolution was discovered at 77 K. Deconvolution of the fluorescence decay curve was then used to reveal the energy transfer channels and the associated transfer rates. Except for the fluorescence lifetimes of terminal emitters, four time components, i.e., 9 ps, 13 ps, 23 ps, and 55 ps, were recognized in the energy transfer in the PBSs. The energy transfer dynamics in the PBSs were further analyzed by combining the cryo-EM structure and the spectral properties in detail. The findings from this study aid in our understanding of the energy transfer mechanisms in PBSs.

Identification of a Ubiquinone–Ubiquinol Quinhydrone Complex in Bacterial Photosynthetic Membranes and Isolated Reaction Centers by Time-Resolved Infrared Spectroscopy

Abstract: Ubiquinone redox chemistry is of fundamental importance in biochemistry, notably in bioenergetics. The bi-electronic reduction of ubiquinone to ubiquinol has been widely studied, including by Fourier transform infrared (FTIR) difference spectroscopy, in several systems. In this paper, we have recorded static and time-resolved FTIR difference spectra reflecting light-induced ubiquinone reduction to ubiquinol in bacterial photosynthetic membranes and in detergent-isolated photosynthetic bacterial reaction centers. We found compelling evidence that in both systems under strong light illumination—and also in detergent-isolated reaction centers after two saturating flashes—a ubiquinone–ubiquinol charge-transfer quinhydrone complex, characterized by a characteristic band at ~1565 cm−1, can be formed. Quantum chemistry calculations confirmed that such a band is due to formation of a quinhydrone complex. We propose that the formation of such a complex takes place when Q and QH2 are forced, by spatial constraints, to share a common limited space as, for instance, in detergent micelles, or when an incoming quinone from the pool meets, in the channel for quinone/quinol exchange at the QB site, a quinol coming out. This latter situation can take place both in isolated and membrane bound reaction centers Possible consequences of the formation of this charge-transfer complex under physiological conditions are discussed.