Oxygen-evolving Photosynthetic Organisms Research Articles

Photosystem I: A Paradigm for Understanding Biological Environmental Adaptation Mechanisms in Cyanobacteria and Algae.

The process of oxygenic photosynthesis is primarily driven by two multiprotein complexes known as photosystem II (PSII) and photosystem I (PSI). PSII facilitates the light-induced reactions of water-splitting and plastoquinone reduction, while PSI functions as the light-driven plastocyanin-ferredoxin oxidoreductase. In contrast to the highly conserved structure of PSII among all oxygen-evolving photosynthetic organisms, the structures of PSI exhibit remarkable variations, especially for photosynthetic organisms that grow in special environments. In this review, we make a concise overview of the recent investigations of PSI from photosynthetic microorganisms including prokaryotic cyanobacteria and eukaryotic algae from the perspective of structural biology. All known PSI complexes contain a highly conserved heterodimeric core; however, their pigment compositions and peripheral light-harvesting proteins are substantially flexible. This structural plasticity of PSI reveals the dynamic adaptation to environmental changes for photosynthetic organisms.

Seasonal Monitoring Of Chlorophyll-A With Landsat 8 Oli In The Madura Strait, Pasuruan, East Java, Indonesia

Chlorophyll-a (Chl-a) is a type of pigment is most common and predominant in all oxygen-evolving photosynthetic organisms such as higher plants, red and green algae. The concentrations of high chlorophyll-a (Chl-a) in coastal waters tend to be lower offshore due to land through river water runoff. The Madura Strait is one of the Indonesian basins that is widely used for fisheries activity, which directly impacts and puts quite high pressure on the aquatic resources. In addition, the development of urban areas and changes of land use in the hinterland areas of East Java Province due to increasing population are also intensive. The objectives of this research were: (1) to map the distribution of chlorophyll-a, its concentration and dynamics in the Madura Strait near the Pasuruan coastal area using remote sensing for both dry and rainy seasons, (2) figure out the influence of rivers or other oceanographic factors that may occur, and (3) calculate the accuracy of the estimation compared to the field data. The Landsat 8 OLI imagery was used to determine the concentration of Chl-a and analyze its seasonal spatial distribution pattern. The results show that (1) spatial distribution of chlorophyll-a (Chl-a), its concentration and dynamics in the Madura Strait waters near the Pasuruan coastal area varies between dry and rainy months or seasons, (2) input from rivers, waves, tidal level, and eddy circulation constitute the oceanographic parameters that influence the spatial distribution pattern of chlorophyll-a (Chl-a) in the Madura Strait waters near the Pasuruan coastal area, and (3) validation of the estimated Chl-a concentrations from Landsat 8 OLI using field data has shown RMSE value of 0.49.

Color Sensing and Signal Transmission Diversity of Cyanobacterial Phytochromes and Cyanobacteriochromes.

To perceive fluctuations in light quality, quantity, and timing, higher plants have evolved diverse photoreceptors including UVR8 (a UV-B photoreceptor), cryptochromes, phototropins, and phytochromes (Phys). In contrast to plants, prokaryotic oxygen-evolving photosynthetic organisms, cyanobacteria, rely mostly on bilin-based photoreceptors, namely, cyanobacterial phytochromes (Cphs) and cyanobacteriochromes (CBCRs), which exhibit structural and functional differences compared with plant Phys. CBCRs comprise varying numbers of light sensing domains with diverse color-tuning mechanisms and signal transmission pathways, allowing cyanobacteria to respond to UV-A, visible, and far-red lights. Recent genomic surveys of filamentous cyanobacteria revealed novel CBCRs with broader chromophore-binding specificity and photocycle protochromicity. Furthermore, a novel Cph lineage has been identified that absorbs blue-violet/yellow-orange light. In this minireview, we briefly discuss the diversity in color sensing and signal transmission mechanisms of Cphs and CBCRs, along with their potential utility in the field of optogenetics.

Triplet–triplet energy transfer in artificial and natural photosynthetic antennas

In photosynthetic organisms, protection against photooxidative stress due to singlet oxygen is provided by carotenoid molecules, which quench chlorophyll triplet species before they can sensitize singlet oxygen formation. In anoxygenic photosynthetic organisms, in which exposure to oxygen is low, chlorophyll-to-carotenoid triplet-triplet energy transfer (T-TET) is slow, in the tens of nanoseconds range, whereas it is ultrafast in the oxygen-rich chloroplasts of oxygen-evolving photosynthetic organisms. To better understand the structural features and resulting electronic coupling that leads to T-TET dynamics adapted to ambient oxygen activity, we have carried out experimental and theoretical studies of two isomeric carotenoporphyrin molecular dyads having different conformations and therefore different interchromophore electronic interactions. This pair of dyads reproduces the characteristics of fast and slow T-TET, including a resonance Raman-based spectroscopic marker of strong electronic coupling and fast T-TET that has been observed in photosynthesis. As identified by density functional theory (DFT) calculations, the spectroscopic marker associated with fast T-TET is due primarily to a geometrical perturbation of the carotenoid backbone in the triplet state induced by the interchromophore interaction. This is also the case for the natural systems, as demonstrated by the hybrid quantum mechanics/molecular mechanics (QM/MM) simulations of light-harvesting proteins from oxygenic (LHCII) and anoxygenic organisms (LH2). Both DFT and electron paramagnetic resonance (EPR) analyses further indicate that, upon T-TET, the triplet wave function is localized on the carotenoid in both dyads.

The amino-terminal conserved domain of 4-hydroxy-3-methylbut-2-enyl diphosphate reductase is critical for its function in oxygen-evolving photosynthetic organisms

4-hydroxy-3-methylbut-2-enyl diphosphate reductase (HDR), also known as isoprenoid synthesis H (IspH) or lysis-tolerant B (LytB), catalyzes the last step of the methylerythritol phosphate pathway to synthesize isopentenyl diphosphate and dimethylallyl diphosphate. The structure and reaction mechanism of IspH have been actively investigated in Escherichia coli but little is known in plants. Compared with the bacterial IspH, cyanobacterial and plant HDRs all contain an extra N-terminal conserved domain (NCD) that is essential for their function. Tyr72 in the NCD and several plant-specific residues around the central active site are critical for Arabidopsis HDR function. These results suggest that the structure and reaction mechanism of HDR/IspH may be different between plants and bacteria. The E. coli IspH is an iron-sulfur protein that is sensitive to oxygen. It is possible that the cyanobacterial HDR may independently evolve from the common ancestor of prokaryotes to obtain the NCD, which may protect the enzyme from high concentration of oxygen during photosynthesis.

Photosystem I Reduction in Diatoms: As Complex as the Green Lineage Systems but Less Efficient

Diatoms occupy a key branch in the evolutionary tree of oxygen-evolving photosynthetic organisms. Here, the electron transfer reaction mechanism from cytochrome c₆ to photosystem I from the diatom Phaeodactylum tricornutum has been analyzed by laser-flash absorption spectroscopy. Kinetic traces of photosystem I reduction fit to biphasic curves, the analysis of the observed rate constants indicating that electron transfer occurs in a cytochrome c₆/photosystem I transient complex, which undergoes a reorganization process from the initial encounter complex to the optimized final configuration. The mild ionic strength dependence of the rate constants makes evident the relatively weak electrostatically attractive nature of the interaction. Taken together, these results indicate that the "red" Phaeodactylum system is less efficient than "green" systems, both in the formation of the properly arranged (cytochrome c₆/photosystem I) complex and in the electron transfer itself. The results obtained from cross-reactions with cytochrome c₆ and photosystem I from cyanobacteria, green algae, and plants shed light on the different evolutionary pathway of the electron transfer to photosystem I in diatoms with regard to the way that it evolved in higher plants.

Light Harvesting and Blue-Green Light Induced Non-Photochemical Quenching in Two Different C-Phycocyanin Mutants of Synechocystis PCC 6803

Cyanobacteria are oxygen-evolving photosynthetic organisms that harvest sunlight and convert excitation energy into chemical energy. Most of the light is absorbed by large light harvesting complexes called phycobilisomes (PBs). In high-light conditions, cyanobacteria switch on a photoprotective mechanism called non-photochemical quenching (NPQ): During this process, absorption of blue-green light transforms the inactive orange form of the orange carotenoid protein OCP (OCP(o)) into the red active form OCP(r) that subsequently binds to the PB, resulting in a substantial loss of excitation energy and corresponding decrease of the fluorescence. In wild-type cells, the quenching site is a bilin chomophore that fluoresces at 660 nm and which is called APC(Q)(660). In the present work, we studied NPQ in two different types of mutant cells (CB and CK) that possess significantly truncated PBs, using spectrally resolved picosecond fluorescence spectroscopy. The results are in very good agreement with earlier in vitro experiments on quenched and unquenched PBs, although the fraction of quenched PBs is far lower in vivo. It is also lower than the fraction of PBs that is quenched in wild-type cells, but the site, rate, and location of quenching appear to be very similar.

A kinetic model of rapidly reversible nonphotochemical quenching

Oxygen-evolving photosynthetic organisms possess nonphotochemical quenching (NPQ) pathways that protect against photo-induced damage. The majority of NPQ in plants is regulated on a rapid timescale by changes in the pH of the thylakoid lumen. In order to quantify the rapidly reversible component of NPQ, called qE, we developed a mathematical model of pH-dependent quenching of chlorophyll excitations in Photosystem II. Our expression for qE depends on the protonation of PsbS and the deepoxidation of violaxanthin by violaxanthin deepoxidase. The model is able to simulate the kinetics of qE at low and high light intensities. The simulations suggest that the pH of the lumen, which activates qE, is not itself affected by qE. Our model provides a framework for testing hypothesized qE mechanisms and for assessing the role of qE in improving plant fitness in variable light intensity.

Structural snapshots along the reaction pathway of ferredoxin–thioredoxin reductase

Oxygen-evolving photosynthetic organisms regulate carbon metabolism through a light-dependent redox signalling pathway. Electrons are shuttled from photosystem I by means of ferredoxin (Fdx) to ferredoxin-thioredoxin reductase (FTR), which catalyses the two-electron-reduction of chloroplast thioredoxins (Trxs). These modify target enzyme activities by reduction, regulating carbon flow. FTR is unique in its use of a [4Fe-4S] cluster and a proximal disulphide bridge in the conversion of a light signal into a thiol signal. We determined the structures of FTR in both its one- and its two-electron-reduced intermediate states and of four complexes in the pathway, including the ternary Fdx-FTR-Trx complex. Here we show that, in the first complex (Fdx-FTR) of the pathway, the Fdx [2Fe-2S] cluster is positioned suitably for electron transfer to the FTR [4Fe-4S] centre. After the transfer of one electron, an intermediate is formed in which one sulphur atom of the FTR active site is free to attack a disulphide bridge in Trx and the other sulphur atom forms a fifth ligand for an iron atom in the FTR [4Fe-4S] centre--a unique structure in biology. Fdx then delivers a second electron that cleaves the FTR-Trx heterodisulphide bond, which occurs in the Fdx-FTR-Trx complex. In this structure, the redox centres of the three proteins are aligned to maximize the efficiency of electron transfer from the Fdx [2Fe-2S] cluster to the active-site disulphide of Trxs. These results provide a structural framework for understanding the mechanism of disulphide reduction by an iron-sulphur enzyme and describe previously unknown interaction networks for both Fdx and Trx (refs 4-6).

Purification of plastocyanin and cytochrome c6 from plants, green algae, and cyanobacteria.

Plastocyanin and cytochrome c6 are widely distributed over the oxygen-evolving photosynthetic organisms. The two proteins are functionally equivalent, but strongly differ in their global electrostatic charge. In fact, they are acidic in eukaryotes, but either neutral or basic in cyanobacteria. The difference in their electrostatic features is a critical factor in designing the purification procedure, which must be modified and adapted accordingly. This chapter reports the methods for producing (including cell cultures), isolating and purifying plastocyanin and cytochrome c6-which greatly differ in their isoelectric point-from a number of eukaryotic and prokaryotic organisms.

Proline 14 is impairing the electron transfer from plastocyanin to photosystem I in the prochlorophyte Prochloorothrix hollandica

Plastocyanin (Pc) from Prochlorothrix hollandica - which occupies a unique divergent branch in the evolutionary tree of oxygen-evolving photosynthetic organisms - exhibits some peculiar differences at its north hydrophobic patch as compared with Pc from other organisms. Actually, residues at positions 12 and 14 are tyrosine and proline, respectively, in Prochlorothrix but they are glycine and leucine in all others. Photosystem I (PSI) reduction by single (Y12G, Y12F, Y12W and P14L) and double (Y12G/P14L) mutants of Prochlorothrix Pc has been investigated by laser flash absorption spectroscopy. With all the mutants, the observed first-order rate constant reaches a saturation plateau at high Pc concentration, as is the case with WT Pc. This finding suggests the formation of a transient Pc-PSI complex. Such a complex appears to be hydrophobic in nature as the observed rate constant is independent of ionic strength. The association constant for complex formation is not significantly altered by mutations, but the electron transfer rate constant is drastically changed, in particular by mutations at position 14. In fact, replacement of Pro-14 with leucine makes the rate constant for electron transfer with the mutant three times higher than that with the WT molecule. Pc mutants have indeed been used as donor proteins in experiments with PSI isolated from different organisms (cyanobacteria and higher plants); the resulting data reveal that reversion of the exclusive Pro-14 of Prochlorothrix Pc to the standard leucine enhances the reactivity of Pc towards PSI.

Time-resolved monitoring of flash-induced changes of fluorescence quantum yield and decay of delayed light emission in oxygen-evolving photosynthetic organisms.

The present contribution describes a new experimental setup that permits time-resolved monitoring of the rise kinetics of the relative fluorescence yield, Phi(rel)(t), and simultaneously of the decay of delayed light emission, L(t), induced by strong actinic laser flashes. The results obtained by excitation of dark-adapted samples with a train of eight flashes reveal (a) in suspensions of spinach thylakoids, Phi(rel)(t) exhibits a typical period four oscillation that is characteristic for a dependence on the redox states S(i)() of the water oxidizing complex (WOC), (b) the relative extent of the unresolved "instantaneous" rise to the level (100 ns) at 100 ns and the maximum values of Phi(rel)(t) attained at about 45 s after each actinic flash, (45 s) synchronously oscillate and exhibit the largest values at flash nos. 1 and 5 and minima after flash nos. 2 and 3, (c) opposite effects are observed for the normalized extent of the rise kinetics in the 100 ns to 5 s time domain of relative fluorescence yield, Phi(rel)(5 s) - Phi(rel)(100 ns), i.e., both parameters attain minimum and maximum values after the first/fifth and second/third flash, respectively, and (d) analogous features for the "fast" and "slow" ns-kinetics of the fluorescence rise were observed in suspensions of Chlamydomas reinhardtii cells. A slight phase shift by one flash is ascribed to physiological differences. The applicability of this noninvasive technique to study reactions of photosystem II, especially the reduction kinetics of P680(*)(+) and their dependence on the redox state S(i)() of the WOC, is discussed.

The solution structure of photosystem I accessory protein E from the cyanobacterium Nostoc sp. strain PCC 8009.

PsaE is a small basic subunit located on the stromal (cytoplasmic) side of photosystem I. In cyanobacteria, this subunit participates in cyclic electron transport and modulates the interactions of the complex with soluble ferredoxin. The PsaE protein isolated from the cyanobacterium Synechococcus sp. strain PCC 7002 adopts the beta topology of an SH3 domain, with five beta strands (betaA through betaE) and a turn of 3(10) helix between strands betaD and betaE [Falzone, C. J., Kao, Y.-H., Zhao, J., Bryant, D. A., and Lecomte, J. T. J. (1994) Biochemistry 33, 6052-6062]. The primary structure of the PsaE protein is strongly conserved across all oxygen-evolving photosynthetic organisms. However, variability in loop lengths, as well as N- or C-terminal extensions, suggests that the structure of a second representative PsaE subunit would be useful to characterize the interactions among photosystem I polypeptides. In this work, the solution structure of PsaE from the cyanobacterium Nostoc sp. strain PCC 8009 was determined by NMR methods. Compared to PsaE from Synechococcus sp. strain PCC 7002, this PsaE has a seven-residue deletion in the loop connecting strands betaC and betaD, and an eight-residue C-terminal extension. Angular and distance restraints derived from homonuclear and heteronuclear NMR experiments were used to calculate structures by a distance-geometry/simulated-annealing protocol. A family of 20 structures (rmsd of 0.24 A in the regular secondary structure) is presented. Differences between the two cyanobacterial proteins are mostly confined to the CD loop region; the C-terminal extension is disordered. The thermodynamic stability of Nostoc sp. strain PCC 8009 PsaE toward urea denaturation was measured by circular dichroism and fluorescence spectroscopy, and thermal denaturation was monitored by UV absorption spectroscopy. Chemical and thermal denaturation curves are modeled satisfactorily with two-state processes. The DeltaG degrees of unfolding at room temperature is 12.4 +/- 0.3 kJ mol(-1) (pH 5), and the thermal transition midpoint is 59 +/- 1 degrees C (pH 7). Interactions with other proteins in the photosystem I complex may aid in maintaining PsaE in its native state under physiological conditions.

The reaction mechanism of Photosystem I reduction by plastocyanin and cytochrome c 6 follows two different kinetic models in the cyanobacterium Pseudanabaena sp. PCC 6903

Plastocyanin (Pc) and cytochrome c6 (Cyt) have been purified to homogeneity from the cyanobacterium Pseudanabaena sp. PCC 6903, which occupies a unique divergent branch in the evolutionary tree of oxygen-evolving photosynthetic organisms. The two metalloproteins have similar molecular masses (9–10 kDa), as well as almost identical isoelectric points (ca. 8) and midpoint redox potentials (ca. 350 mV, at pH 7). Their reaction mechanism of electron transfer to Photosystem I (PS I) has been analyzed by laser-flash absorption spectroscopy. The kinetic traces with Pc correspond to monophasic kinetics, whereas those with Cyt are better fitted to biphasic curves. The observed pseudo first-order rate constant (kobs) with Pc and that for the slower phase with Cyt exhibit saturation profiles at increasing donor protein concentrations, thereby suggesting that the two metalloproteins are able to form transient complexes with PS I. The ionic strength dependence of the rate constants for complex formation makes evident the electrostatic nature of intermediate complexes. The experimental findings indicate that the PS I reduction kinetics in Pseudanabaena follow a type II mechanism with Pc and a type III mechanism with Cyt, according to the different kinetic models proposed previously [(Hervas M, Navarro JA, Diaz A, Bottin H and De la Rosa MA (1995) Biochemistry 34: 11321–11326)]. From an evolutionary point of view, this reinforces our previous observation that PS I was first adapted to operate efficiently with positively charged Cyt rather than with Pc.

THE CHLOROPHYLL-CAROTENOID PROTEINS OF OXYGENIC PHOTOSYNTHESIS.

The chlorophyll-carotenoid binding proteins responsible for absorption and conversion of light energy in oxygen-evolving photosynthetic organisms belong to two extended families: the Chl a binding core complexes common to cyanobacteria and all chloroplasts, and the nuclear-encoded light-harvesting antenna complexes of eukaryotic photosynthesizers (Chl a/b, Chl a/c, and Chl a proteins). There is a general consensus on polypeptide and pigment composition for higher plant pigment proteins. These are reviewed and compared with pigment proteins of chlorophyte, rhodophyte, and chromophyte algae. Major advances have been the determination of the structures of LHCII (major Chl a/b complex of higher plants), cyanobacterial Photosystem I, and the peridinen-Chl a protein of dinoflagellates to atomic resolution. Better isolation methods, improved transformation procedures, and the availability of molecular structure models are starting to provide insights into the pathways of energy transfer and the macromolecular organization of thylakoid membranes.

Excited-state redox potentials and the Z scheme of photosynthesis

The Z scheme for electron flow in oxygen-evolving photosynthetic organisms was first proposed 25 years ago by R. Hill and F. BendalP. Their original diagram is shown in Fig. 1. The basic idea, that two photochemical systems are arranged in series so that two photons act sequentially to drive electrons from the very poor electron donor 1-I20 to the weak electron acceptor NADP, is now embraced by essentially every worker in the field. This scheme rightly stands as the central integrating construct for all work on electron transport in oxygenevolving photosynthetic organisms. Introductory biochemistry texts invariably contain a version of the scheme, usually with an explanation of some of the classic experiments (such as the red drop and enhancement effects) that led to the acceptance of the idea of a sequence of two photochemical steps, each with a somewhat different wavelength optimum. The Z scheme is really a synthesis of thermodynamic and kinetic data. The midpoint redox potentials of the various electron carriers are now usually plotted vertically so that more reducing compounds are above more oxidizing ones. This ensures that thermodynamically favorable reactions are 'downhill' on the diagram. The horizontal axis corresponds to a reaction sequence and the kinetically important reactions are connected by arrows. Unfortunately, many authors are extremely careless about constructing their Z schemes, so much so that the understanding of what is in reality a simple concept is greatly impeded. Several Z schemes in the literature have P~0, the reaction centerchlorophyll of photosystem II, at an E m (oxidation-reduction midpoint potential) of +450 mV, and show electrons flowing uphill to it from 1420 against a

Primary electron acceptor complex of photosystem I in spinach chloroplasts

PHOTOSYSTEM I (PSI) is the photochemical reaction complex involved in the generation of a low potential reductant in oxygen-evolving photosynthetic organisms. The primary photochemical reaction in PSI has been identified as the photo-oxidation of a reaction centre chlorophyll complex P700 (ref. 1). This photo-oxidation occurs both at room temperature and in the frozen state at temperatures as low as 4.2 K. At cryogenic temperatures this photo-oxidation has generally been found to be irreversible2. Malkin and Bearden3 showed that the electron acceptor for this photo-oxidation reaction is a bound ferredoxin. This ferredoxin has two iron–sulphur centres with very low redox potentials (Em10 = −550 and −590 mV)4–6. It has been proposed that these centres are the primary electron acceptor of PSI3,4. More recent evidence suggests it may not be7–11, and we have shown11 that when the bound ferredoxin is chemically reduced, illumination at low temperature results in the photo-oxidation of P700 without any change in the bound ferredoxin, this oxidation is reversible. In photosynthetic bacteria a component with an EPR signal at g = 1.82 has been identified as the primary electron acceptor of the photochemical system12,13 and it is possible that a similar component might be involved in chloroplast photochemical reactions. McIntosh et al.14 obtained kinetic evidence for the presence in PSI particles of a component with an EPR signal at g = 2.06 and g = 1.76 which showed a reversible redox change on illumination of the particles at low temperature parallel to that of P700. The changes observed were small and it was not possible to obtain a spectrum of the component. Using a more highly purified PSI preparation we have now obtained strong evidence that this component is the primary electron acceptor of PSI.