Electron Acceptor Of Photosystem Research Articles

Potassium alleviates over-reduction of the photosynthetic electron transport chain and helps to maintain photosynthetic function under salt-stress.

Potassium ions enhance photosynthetic tolerance to salt stress. We hypothesized that potassium ions, by minimizing the trans-thylakoid proton diffusion potential difference, can alleviate over-reduction of the photosynthetic electron transport chain and maintain the functionality of the photosynthetic apparatus. This study investigated the effects of exogenous potassium on the transcription level and activity of proteins related to the photosynthetic electron-transport chain of tobacco seedlings under salt stress. Salt stress retarded the growth of seedlings and caused an outflow of potassium ions from the chloroplast. It also lowered qP (indicator of the oxidation state of QA , the primary quinone electron acceptor in Photosystem II (PSII) and YPSII (average photochemical yield of PSII in the light-adapted state) while increasing YNO+NF (nonregulatory energy dissipation in functional and nonfunctional PSII), accompanied by decreased expression of most light-harvesting, energy-transduction, and electron-transport genes. However, exogenous potassium prevented these effects due to NaCl. Interestingly, lincomycin (an inhibitor of the synthesis of chloroplast-encoded proteins in PSII) significantly diminished the alleviation effect of exogenous potassium on salt stress. We attribute the comprehensive NaCl-induced downregulation of transcription and photosynthetic activities to retrograde signaling induced by reactive oxygen species. There probably exist at least two types of retrograde signaling induced by reactive oxygen species, distinguished by their sensitivity to lincomycin. Exogenous potassium appears to exert its primary effect by ameliorating the trans-thylakoid proton diffusion potential difference via a potassium channel, thereby accelerating ATP synthesis and carbon assimilation, alleviating over-reduction of the photosynthetic electron transport chain, and maintaining the functionality of photosynthetic proteins.

Structural insights into the action mechanisms of artificial electron acceptors in photosystem II

Photosystem II (PSII) utilizes light energy to split water, and the electrons extracted from water are transferred to QB, a plastoquinone molecule bound to the D1 subunit of PSII. Many artificial electron acceptors (AEAs) with molecular structures similar to that of plastoquinone can accept electrons from PSII. However, the molecular mechanism by which AEAs act on PSII is unclear. Here, we solved the crystal structure of PSII treated with three different AEAs, 2,5-dibromo-1,4-benzoquinone, 2,6-dichloro-1,4-benzoquinone, and 2-phenyl-1,4-benzoquinone, at 1.95 to 2.10Å resolution. Our results show that all AEAs substitute for QB and are bound to the QB-binding site (QB site) to receive electrons, but their binding strengths are different, resulting in differences in their efficiencies to accept electrons. The acceptor 2-phenyl-1,4-benzoquinone binds most weakly to the QB site and showed the highest oxygen-evolving activity, implying a reverse relationship between the binding strength and oxygen-evolving activity. In addition, a novel quinone-binding site, designated the QD site, was discovered, which is located in the vicinity of QB site and close to QC site, a binding site reported previously. This QD site is expected to play a role as a channel or a storage site for quinones to be transported to the QB site. These results provide the structural basis for elucidating the actions of AEAs and exchange mechanism of QB in PSII and also provide information for the design of more efficient electron acceptors.

Ultrafast Electron Transfer in Au–Cyanobacteria Hybrid for Solar to Chemical Production

The rise of inorganic–biohybrid organisms for solar-to-chemical production has spurred mechanistic investigations into the dynamics of the biotic–abiotic interface to drive the development of next-generation hybrid systems. The model system, cyanobacteria–gold nanoparticle hybrids, combines a light harvester with a photosynthetic bacterium to drive the reduction of CO2 to glycerol with improved efficiency and increased glycerol production by 14.6%, in comparison to cyanobacteria only. In this work, we report insights into this unique photochemical behavior and propose a charge-transfer pathway from Au nanoparticle to cyanobacteria. Transient absorption (TA) spectroscopy revealed that photoexcited electron transfer rates are on the order of a few ps to the potential electron acceptor in photosystem II. This work represents a promising platform to utilize a conventional spectroscopic methodology to extract insights from more complex biotic–abiotic hybrid systems.

Acetylacetone Interferes with Carbon and Nitrogen Metabolism of Microcystis aeruginosa by Cutting Off the Electron Flow to Ferredoxin.

The regulation of photosynthetic machinery with a nonoxidative approach is a powerful but challenging strategy for the selective inhibition of bloom-forming cyanobacteria. Acetylacetone (AA) was recently found to be a target-selective cyanocide for Microcystis aeruginosa, but the cause and effect in the studied system are still unclear. By recording of the chemical fingerprints of the cells at two treatment intervals (12 and 72 h with 0.1 mM AA) with omics assays, the molecular mechanism of AA in inactivating Microcystis aeruginosa was elucidated. The results clearly reveal the effect of AA on ferredoxin and the consequent effects on the physiological and biochemical processes of Microcystis aeruginosa. In addition to its role as an electron acceptor of photosystem I, ferredoxin plays pivotal roles in the assimilation of nitrogen in cyanobacterial cells. The effect of AA on ferredoxin and on nonheme iron of photosystem II first cut off the photosynthetic electron transfer flow and then interrupted the synthesis of adenosine triphosphate (ATP) and reduced nicotinamide adenine dinucleotide phosphate (NADPH), which ultimately might affect carbon fixation and nitrogen assimilation metabolisms. The results here provide missing pieces in the current knowledge on the selective inhibition of cyanobacteria, which should shed light on the better control of harmful blooms.

Redox properties and regulatory mechanism of the iron-quinone electron acceptor in photosystem II as revealed by FTIR spectroelectrochemistry.

Photosystem II (PSII) performs oxidation of water and reduction of plastoquinone through light-induced electron transfer. Electron transfer reactions at individual redox cofactors are controlled by their redox potentials, and the forward and backward electron flows in PSII are regulated by tuning them. It is, thus, crucial to accurately estimate the redox potentials of the cofactors and their shifts by environmental changes to understand the regulatory mechanisms in PSII. Fourier-transform infrared (FTIR) spectroelectrochemistry combined with a light-induced difference technique is a powerful method to investigate the mechanisms of the redox reactions in PSII. In this review, we introduce the methodology and the application of this method in the studies of the iron-quinone complex, which consists of two plastoquinone molecules, QA and QB, and the non-heme iron, on the electron-acceptor side of PSII. It is shown that FTIR spectroelectrochemistry is a useful method not only for estimating the redox potentials but also for detecting the reactions of nearby amino-acid residues coupled with the redox reactions.

Fine-tuning of FeS proteins monitored via pulsed EPR redox potentiometry at Q-band

As essential electron translocating proteins in photosynthetic organisms, multiple plant-type ferredoxin (Fdx) isoforms are involved in a high number of reductive metabolic processes in the chloroplast. To allow quick cellular responses under changing environmental conditions, different plant-type Fdxs in Chlamydomonas reinhardtii were suggested to have adapted their midpoint potentials to a wide range of interaction partners. We performed pulsed electron paramagnetic resonance (EPR) monitored redox potentiometry at Q-band on three Fdx isoforms for a straightforward determination of their midpoint potentials. Additionally, site-directed mutagenesis was used to tune the midpoint potential of CrFdx1 in a range of approximately −338 to −511 mV, confirming the importance of single positions in the protein environment surrounding the [2Fe2S] cluster. Our results present a new target for future studies aiming to modify the catalytic activity of CrFdx1 that plays an essential role either as electron acceptor of photosystem I or as electron donor to hydrogenases under certain conditions. Additionally, the precisely determined redox potentials in this work using pulsed EPR demonstrate an alternative method that provides additional advantages compared with the well-established continuous wave EPR technique.

Maximum fluorescence and electron transport kinetics determined by light-induced fluorescence transients (LIFT) for photosynthesis phenotyping

Photosynthetic phenotyping requires quick characterization of dynamic traits when measuring large plant numbers in a fluctuating environment. Here, we evaluated the light-induced fluorescence transient (LIFT) method for its capacity to yield rapidly fluorometric parameters from 0.6 m distance. The close approximation of LIFT to conventional chlorophyll fluorescence (ChlF) parameters is shown under controlled conditions in spinach leaves and isolated thylakoids when electron transport was impaired by anoxic conditions or chemical inhibitors. The ChlF rise from minimum fluorescence (Fo) to maximum fluorescence induced by fast repetition rate (Fm−FRR) flashes was dominated by reduction of the primary electron acceptor in photosystem II (QA). The subsequent reoxidation of QA− was quantified using the relaxation of ChlF in 0.65 ms (Fr1) and 120 ms (Fr2) phases. Reoxidation efficiency of QA− (Fr1/Fv, where Fv = Fm−FRR − Fo) decreased when electron transport was impaired, while quantum efficiency of photosystem II (Fv/Fm) showed often no significant effect. ChlF relaxations of the LIFT were similar to an independent other method. Under increasing light intensities, Fr2′/Fq′ (where Fr2′ and Fq′ represent Fr2 and Fv in the light-adapted state, respectively) was hardly affected, whereas the operating efficiency of photosystem II (Fq′/Fm′) decreased due to non-photochemical quenching. Fm−FRR was significantly lower than the ChlF maximum induced by multiple turnover (Fm−MT) flashes. However, the resulting Fv/Fm and Fq′/Fm′ from both flashes were highly correlated. The LIFT method complements Fv/Fm with information about efficiency of electron transport. Measurements in situ and from a distance facilitate application in high-throughput and automated phenotyping.

FTIR spectroelectrochemistry combined with a light-induced difference technique: Application to the iron-quinone electron acceptor in photosystem II

Photosystem II (PSII) in plants and cyanobacteria performs light-driven water oxidation to obtain electrons necessary for CO2 fixation. In PSII, a series of electron transfer reactions take place from the Mn4CaO5 cluster, the catalytic site of water oxidation, to a plastoquinone molecule via several redox cofactors. Light-induced Fourier transform infrared (FTIR) difference spectroscopy has been extensively used to investigate the structures and reactions of the redox cofactors in PSII. Recently, FTIR spectroelectrochemistry combined with the light-induced difference technique was applied to study the mechanism of electrontransfer regulation in PSII involving the quinone electron acceptors, QA and QB, and the non-heme iron that bridges them. In this mini-review, this combined FTIR method is introduced, and obtained results about the redox reactions of the non-heme iron and QB, involving the long-range interaction of the Mn4CaO5 cluster with the electron-acceptor side, are summarized.

Unique chlorophylls in picoplankton Prochlorococcus sp. "Physicochemical properties of divinyl chlorophylls, and the discovery of monovinyl chlorophyll b as well as divinyl chlorophyll b in the species Prochlorococcus NIES-2086".

In this review, we introduce our recent studies on divinyl chlorophylls functioning in unique marine picoplankton Prochlorococcus sp. (1) Essential physicochemical properties of divinyl chlorophylls are compared with those of monovinyl chlorophylls; separation by normal-phase and reversed-phase high-performance liquid chromatography with isocratic eluent mode, absorption spectra in four organic solvents, fluorescence information (emission spectra, quantum yields, and life time), circular dichroism spectra, mass spectra, nuclear magnetic resonance spectra, and redox potentials. The presence of a mass difference of 278 in the mass spectra between [M+H]+ and the ions indicates the presence of a phytyl tail in all the chlorophylls. (2) Precise high-performance liquid chromatography analyses show divinyl chlorophyll a' and divinyl pheophytin a as the minor key components in four kinds of Prochlorococcus sp.; neither monovinyl chlorophyll a' nor monovinyl pheophytin a is detected, suggesting that the special pair in photosystem I and the primary electron acceptor in photosystem II are not monovinyl but divinyl-type chlorophylls. (3) Only Prochlorococcus sp. NIES-2086 possesses both monovinyl chlorophyll b and divinyl chlorophyll b, while any other monovinyl-type chlorophylls are absent in this strain. Monovinyl chlorophyll b is not detected at all in the other three strains. Prochlorococcus sp. NIES-2086 is the first example that has both monovinyl chlorophyll b as well as divinyl chlorophylls a/b as major chlorophylls.

Adaptive changes in chlorophyll content and photosynthetic features to low light in Physocarpus amurensis Maxim and Physocarpus opulifolius "Diabolo".

The present study aims to investigate the differences in leaf pigment content and the photosynthetic characteristics under natural and low light intensities between the Chinese native Physocarpus amurensis Maxim and the imported Physocarpus opulifolius “Diabolo” from North America. We aim to discuss the responses and the adaptive mechanism of these two cultivars of Physocarpus to a low light environment. The results show that the specific leaf area (SLA) and the chlorophyll content were significantly increased in the leaves of both Physocarpus cultivars in response to a low light intensity, and the SLA and chlorophyll content were higher in the leaves of low light-treated P. opulifolius “Diabolo” compared with the leaves of low light-treated P. amurensis Maxim. Moreover, the content of anthocyanin was markedly reduced in the leaves of P. opulifolius “Diabolo” under low light intensity, which allowed for a greater capacity of photon capture under the low light condition. Under natural light, the photosynthetic carbon assimilation capacity was greater in the leaves of P. amurensis Maxim compared with the leaves of P. opulifolius “Diabolo” that were rich with anthocyanin. However, in response to low light, AQY, Pmax, LCP and LSP decreased to a lesser extent in the leaves of P. opulifolius “Diabolo” compared with the leaves of P. amurensis Maxim. These results suggest that P. opulifolius “Diabolo” exhibits a greater ability in adaption to low light, and it is probably related to the relatively higher chlorophyll content and the smaller SLA in the leaves of P. opulifolius “Diabolo.” In addition, the low light intensity resulted in a reduced photochemical activity of photosystem (PS) II in the leaves of both Physocarpus, as evidenced by increased values of the relative variable fluorescence at point J and point I on the OJIP curve. This result suggests that the electron acceptor in PS II was the major responsive site to the low light stress in the leaves of both Physocarpus cultivars, and that the low light intensity significantly inhibited electron transfer on the acceptor side of PS II and reduced the activity of the oxygen-evolving complex (OEC) in the leaves of both Physocarpus cultivars. The PS II function in P. opulifolius “Diabolo” was higher than that in P. amurensis Maxim in response to low light. Under low light, the composition of photosynthetic pigments was altered in the leaves of P. opulifolius “Diabolo” in order to maintain a relatively high activity of primary photochemical reactions, and this is the basis of the greater photosynthetic carbon assimilation capacity and one of the main reasons for the better shade-tolerance in P. opulifolius “Diabolo.”

Chloramphenicol Mediates Superoxide Production in Photosystem II and Enhances Its Photodamage in Isolated Membrane Particles.

Chloramphenicol (CAP) is an inhibitor of protein synthesis, which is frequently used to decouple photodamage and protein synthesis dependent repair of Photosystem II during the process of photoinhibition. It has been reported earlier that CAP is able to mediate superoxide production by transferring electrons from the acceptor side of Photosystem I to oxygen. Here we investigated the interaction of CAP with Photosystem II electron transport processes by oxygen uptake and variable chlorophyll fluorescence measurements. Our data show that CAP can accept electrons at the acceptor side of Photosystem II, most likely from Pheophytin, and deliver them to molecular oxygen leading to superoxide production. In addition, the presence of CAP enhances photodamage of Photosystem II electron transport in isolated membrane particles, which effect is reversible by superoxide dismutase. It is concluded that CAP acts as electron acceptor in Photosystem II and mediates its superoxide dependent photodamage. This effect has potential implications for the application of CAP in photoinhibitory studies in intact systems.

Modified molecular interactions of the pheophytin and plastoquinone electron acceptors in photosystem II of chlorophyll D-containing Acaryochloris marina as revealed by FTIR spectroscopy.

Acaryochloris marina is a unique cyanobacterium that contains chlorophyll (Chl) d as a major pigment. Because Chl d has smaller excitation energy than Chl a used in ordinary photosynthetic organisms, the energetics of the photosystems of A. marina have been the subject of interest. It was previously shown that the redox potentials (E m's) of the redox-active pheophytin a (Pheo) and the primary plastoquinone electron acceptor (QA) in photosystem II (PSII) of A. marina are higher than those in Chl a-containing PSII, to compensate for the smaller excitation energy of Chl d (Allakhverdiev et al., Proc Natl Acad Sci USA 107: 3924-3929, 2010; ibid. 108: 8054-8058, 2011). To clarify the mechanisms of these E m increases, in this study, we have investigated the molecular interactions of Pheo and QA in PSII core complexes from A. marina using Fourier transform infrared (FTIR) spectroscopy. Light-induced FTIR difference spectra upon single reduction of Pheo and QA showed that spectral features in the regions of the keto and ester C=O stretches and the chlorin ring vibrations of Pheo and in the CO/CC stretching region of the Q A (-) semiquinone anion in A. marina are significantly different from those of the corresponding spectra in Chl a-containing cyanobacteria. These observations indicate that the molecular interactions, including the hydrogen bond interactions at the C=O groups, of these cofactors are modified in their binding sites of PSII proteins. From these results, along with the sequence information of the D1 and D2 proteins, it is suggested that A. marina tunes the E m's of Pheo and QA by altering nearby hydrogen bond networks to modify the structures of the binding pockets of these cofactors.

Photosynthetic acclimation to drought stress in Agave salmiana Otto ex Salm-Dyck seedlings is largely dependent on thermal dissipation and enhanced electron flux to photosystem I.

Agave salmiana Otto ex Salm-Dyck, a crassulacean acid metabolism plant that is adapted to water-limited environments, has great potential for bioenergy production. However, drought stress decreases the requirement for light energy, and if the amount of incident light exceeds energy consumption, the photosynthetic apparatus can be injured, thereby limiting plant growth. The objective of this study was to evaluate the effects of drought and re-watering on the photosynthetic efficiency of A. salmiana seedlings. The leaf relative water content and leaf water potential decreased to 39.6% and -1.1MPa, respectively, over 115 days of water withholding and recovered after re-watering. Drought caused a direct effect on photosystem II (PSII) photochemistry in light-acclimated leaves, as indicated by a decrease in the photosynthetic electron transport rate. Additionally, down-regulation of photochemical activity occurred mainly through the inactivation of PSII reaction centres and an increased thermal dissipation capacity of the leaves. Prompt fluorescence kinetics also showed a larger pool of terminal electron acceptors in photosystem I (PSI) as well as an increase in some JIP-test parameters compared to controls, reflecting an enhanced efficiency and specific fluxes for electron transport from the plastoquinone pool to the PSI terminal acceptors. All the above parameters showed similar levels after re-watering. These results suggest that the thermal dissipation of excess energy and the increased energy conservation from photons absorbed by PSII to the reduction of PSI end acceptors may be an important acclimation mechanism to protect the photosynthetic apparatus from over-excitation in Agave plants.

ChemInform Abstract: Demetalation of Chlorophyll Pigments

AbstractReview: 159 refs.

Demetalation of Chlorophyll Pigments

Natural chlorophylls (Chls) and bacteriochlorophyll (BChls), which are major pigments in photosynthesis, possess a central Mg (or Zn) in their cyclic tetrapyrrole macrocycles. Removal of the central metal atom from chlorophyllous pigments, called pheophytinization, is a biologically important reaction in that it allows production of the primary electron acceptors in photosystem II(-type) reaction centers and is one of the crucial steps in the Chl degradation pathway. Pheophytinization in processed and postharvest foods derived from vegetables and fruits has attracted considerable attention in agricultural and food chemistry from the viewpoints of maintenance of their color level and evaluation by consumers. In this review, we focus on in vivo demetalation reactions and demetalated products of chlorophyllous pigments. In addition, we summarize kinetic studies on in vitro demetalation of natural (B)Chl molecules and their synthetic analogs under acidic conditions.

Structure‐Dependent Demetalation Kinetics of Chlorophyll a Analogs under Acidic Conditions

Demetalation of chlorophyll (Chl) a and its analogs is an important reaction in oxygenic photosynthetic organisms, which produces the primary electron acceptors in photosystem II reaction centers and is crucial in the Chl degradation. From these viewpoints, demetalation reactions of four Chl a analogs, 3,8-divinyl-Chl a (DV-Chl a), 3-devinyl-3-ethyl-Chl a (mesoChl a), 13(2)-demethoxycarbonyl-Chl a (pyroChl a) and protochlorophyll a (PChl a), were kinetically analyzed under weakly acidic conditions, and were compared with that of Chl a. DV-Chl a exhibited slower demetalation kinetics than did Chl a, whereas demetalation of mesoChl a was faster than that of Chl a. The difference in demetalation kinetics of the three chlorophyllous pigments originates from the electron-withdrawing ability of the vinyl group as the peripheral substituent compared with the ethyl group. Removal of the electron-withdrawing and homoconjugating 13(2) -methoxycarbonyl group in Chl a (Chl a → pyroChl a) accelerated demetalation kinetics by two-fold. PChl a possessing the porphyrin-type skeleton exhibited slower demetalation kinetics than Chl a. The structure-dependent demetalation properties of Chl a analogs will be useful for understanding in vivo Chl demetalation reactions in oxygenic photosynthetic organisms.

High and low potential forms of the Q A quinone electron acceptor in Photosystem II of Thermosynechococcus elongatus and spinach

The redox potential of Q A in Photosystem II (PSII) from Thermosynechococcus elongatus was titrated monitoring chlorophyll fluorescence. A high potential form ( E m = +60 ± 25 mV) was found in the absence of Mn 4Ca, the active site for water oxidation. The low potential form ( E m = −60 ± 48 mV), which is difficult to measure in conventional titration experiments, could be “locked in” by cross-linking the active enzyme. This indicates that the presence of Mn 4Ca is relayed to the quinone site by significant structural changes in the protein. The presence of high and low potential forms agrees with what has been seen in plants, algae from our lab and in T. elongatus (Shibamoto et al., Biochemistry 48 (2009) 10682–10684). In the latter work, the potentials of Q A were shifted to lower potentials compared to other measurements. The redox potential of Q A in Mn-depleted PSII from spinach was titrated in the presence of redox mediators and the midpoint potential was shifted by 80 mV towards a more negative value compared to titrations without mediators. The lower values of the midpoint potential of the ( Q A / Q A - ) redox couple in the literature could be due to a perturbation due to a specific mediator.

Time-Resolved FTIR Difference Spectroscopy in Combination with Specific Isotope Labeling for the Study of A 1, the Secondary Electron Acceptor in Photosystem 1

A phylloquinone molecule (2-methyl, 3-phytyl, 1, 4-naphthoquinone) occupies the A 1 binding site in photosystem 1 particles from Synechocystis sp. 6803. In menB mutant photosystem 1 particles from the same species, plastoquinone-9 occupies the A 1 binding site. By incubation of menB mutant photosystem 1 particles in the presence of phylloquinone, it was shown in another study that phylloquinone will displace plastoquinone-9 in the A 1 binding site. We describe the reconstitution of unlabeled ( 16O) and 18O-labeled phylloquinone back into the A 1 binding site in menB photosystem 1 particles. We then produce time-resolved A 1 − / A 1 Fourier transform infrared (FTIR) difference spectra for these menB photosystem 1 particles that contain unlabeled and 18O-labeled phylloquinone. By specifically labeling only the phylloquinone oxygen atoms we are able to identify bands in A 1 − / A 1 FTIR difference spectra that are due to the carbonyl (C O) modes of neutral and reduced phylloquinone. A positive band at 1494 cm −1 in the A 1 − / A 1 FTIR difference spectrum is found to downshift 14 cm −1 and decreases in intensity on 18O labeling. Vibrational mode frequency calculations predict that an antisymmetric vibration of both C O groups of the phylloquinone anion should display exactly this behavior. In addition, phylloquinone that has asymmetrically hydrogen bonded carbonyl groups is also predicted to display this behavior. The positive band at 1494 cm −1 in the A 1 − / A 1 FTIR difference spectrum is therefore due to the antisymmetric vibration of both C O groups of one electron reduced phylloquinone. Part of a negative band at 1654 cm −1 in the A 1 − / A 1 FTIR difference spectrum downshifts 28 cm −1 on 18O labeling. Again, vibrational mode frequency calculations predict this behavior for a C O mode of neutral phylloquinone. The negative band at 1654 cm −1 in the A 1 − / A 1 FTIR difference spectrum is therefore due to a C O mode of neutral phylloquinone. More specifically, calculations on a phylloquinone model molecule with the C 4 O group hydrogen bonded predict that the 1654 cm −1 band is due to the non hydrogen bonded C 1 O mode of neutral phylloquinone.

The nature of the photosystem II reaction centre in the chlorophyll d-containing prokaryote, Acaryochloris marina

Pigment-protein complexes enriched in photosystem II (PS II) have been isolated from the chlorophyll (Chl) d containing cyanobacterium, Acaryochloris marina. A small PS II-enriched particle, we call 'crude reaction centre', contained 20 Chl d, 0.5 Chl a and 1 redox active cytochrome b-559 per 2 pheophytin a, plus the D1 and D2 proteins. A larger PS II-enriched particle, we call 'core', additionally bound the antenna complexes, CP47 and CP43, and had a higher chlorophyll per pheophytin ratio. Pheophytin a could be photoreduced in the presence of a strong reductant, indicating that it is the primary electron acceptor in photosystem II of A. marina. A substoichiometric amount of Chl a (less than one chlorophyll a per 2 pheophytin a) strongly suggests that Chl a does not have an essential role in the photochemistry of PS II in this organism. We conclude that PS II, in A. marina, utilizes Chl d and not Chl a as primary electron donor and that the primary electron acceptor is one of two molecules of pheophytin a.

Structural Basis for the Thermostability of Ferredoxin from the Cyanobacterium Mastigocladus laminosus

Plant-type ferredoxins (Fds) carry a single [2Fe-2S] cluster and serve as electron acceptors of photosystem I (PSI). The ferredoxin from the thermophilic cyanobacterium Mastigocladus laminosus displays optimal activity at 65 degrees C. In order to reveal the molecular factors that confer thermostability, the crystal structure of M.laminosus Fd (mFd) was determined to 1.25 A resolution and subsequently analyzed in comparison with four similar plant-type mesophilic ferredoxins. The topologies of the plant-type ferredoxins are similar, yet two structural determinants were identified that may account for differences in thermostability, a salt bridge network in the C-terminal region, and the flexible L1,2 loop that increases hydrophobic accessible surface area. These conclusions were verified by three mutations, i.e. substitution of L1,2 into a rigid beta-turn ((Delta)L1,2) and two point mutations (E90S and E96S) that disrupt the salt bridge network at the C-terminal region. All three mutants have shown reduced electron transfer (ET) capabilities and [2Fe-2S] stability at high temperatures in comparison to the wild-type mFd. The results have also provided new insights into the involvement of the L1,2 loop in the Fd interactions with its electron donor, the PSI complex.