Crystal structure of the Psb28 accessory factor of Thermosynechococcus elongatus photosystem II at 2.3 Å

Members of the Psb28 family of proteins are accessory factors implicated in the assembly and repair of the photosystem II complex. We present here the crystal structure of the Psb28 protein (Tlr0493) found in the thermophilic cyanobacterium Thermosynechococcus elongatus at a resolution of 2.3 Å. Overall the crystal structure of the Psb28 monomer is similar to the solution structures of C-terminally His-tagged Psb28-1 from Synechocystis sp. PCC 6803 obtained previously by nuclear magnetic resonance spectroscopy. One new aspect is that Escherichia coli-expressed T. elongatus Psb28 is able to form dimers in solution and packs as a dimer of dimers in the crystal. Analysis of wild type and mutant strains of Synechocystis 6803 by blue native-polyacrylamide gel electrophoresis suggests that Psb28-1, the closest homologue to T. elongatus Psb28 in this organism, also exists as an oligomer in vivo, most likely a dimer. In line with the prediction based on the crystal structure of T. elongatus Psb28, the addition of a 3× Flag-tag to the C-terminus of Synechocystis 6803 Psb28-1 interferes with the accumulation of the Psb28-1 oligomer in vivo. In contrast, the more distantly related Psb28-2 protein found in Synechocystis 6803 lacks the residues that stabilize dimer formation in the T. elongatus Psb28 crystal and is detected as a monomer in vivo. Overall our data suggest that the dimer interface in the Psb28 crystal might be physiologically relevant.     

The Antenna System of Photosystem II From Thermosynechococcus elongatus at 3.2 Å Resolution

The content and type of cofactors harboured in the Photosystem II core complex (PS IIcc) of the cyanobacterium Thermosynechococcus elongatus has been determined by biochemical and spectroscopic methods. 17 ± 1 chlorophyll a per pheophytin a and 0.25 β-carotene per chlorophyll a have been found in re-dissolved crystals of dimeric PS IIcc. The X-ray crystal structure of PS IIcc from Thermosynechococcus elongatus at 3.2 Å resolution clearly shows chlorophyll a molecules arranged in two layers close to the cytoplasmic and lumenal sides of the thylakoid membrane. Each of the cytoplasmic layers contains 9 chlorophyll a, whose positions and orientations are related by a local twofold rotation pseudo-C2 axis passing through the non-haem Fe²⁺. These chlorophyll a are arranged comparably to those in the antenna domains of PsaA and PsaB of cyanobacterial Photosystem I affirming an evolutionary relation. The chlorophyll a in the lumenal layer are less well conserved between Photosystems I and II and even between CP43 and CP47 with 4 chlorophyll a in the former and 7 in the latter.     

Sensitivity of the photosynthetic apparatus to UV-A radiation: role of light-harvesting complex II–photosystem II supercomplex organization

In this work we study the effect of UV-A radiation on the function of the photosynthetic apparatus in thylakoid membranes with different organization of the light-harvesting complex II–photosystem II (LHCII–PSII) supercomplex. Leaves and isolated thylakoid membranes from a number of previously characterized pea species with different LHCII size and organization were subjected to UV-A treatment. A relationship was found between the molecular organization of the LHCII (ratio of the oligomeric to monomeric forms of LHCII) and UV-A-induced changes both in the energy transfer from PSII to PSI and between the chlorophyll–protein complexes within the LHCII–PSII supercomplex. Dependence on the organization of the LHCII was also found with regard to the degree of inhibition of the photosynthetic oxygen evolution. The susceptibility of energy transfer and oxygen evolution to UV-A radiation decreased with increasing LHCII oligomerization when the UV-A treatment was performed on isolated thylakoid membranes, in contrast to the effect observed in thylakoid membranes isolated from pre-irradiated pea leaves. The data suggest that UV-A radiation leads mainly to damage of the PSIIα centers. Comparison of membranes with different organization of their LHCII–PSII supercomplex shows that the oligomeric forms of LHCII play a key role for sensitivity to UV-A radiation of the photosynthetic apparatus. S. G. Taneva is Associated member of the Institute of Biophysics, Bulgarian Academy of Sciences.     

Crystal structure of the Psb27 assembly factor at 1.6??: implications for binding to Photosystem II

The biogenesis and oxygen-evolving activity of cyanobacterial Photosystem II (PSII) is dependent on a number of accessory proteins not found in the crystallised dimeric complex. These include Psb27, a small lipoprotein attached to the lumenal side of PSII, which has been assigned a role in regulating the assembly of the Mn(4)Ca cluster catalysing water oxidation. To gain a better understanding of Psb27, we have determined in this study the crystal structure of the soluble domain of Psb27 from Thermosynechococcus elongatus to a resolution of 1.6??. The structure is a four-helix bundle, similar to the recently published solution structures of Psb27 from Synechocystis PCC 6803 obtained by nuclear magnetic resonance (NMR) spectroscopy. Importantly, the crystal structure presented here helps us resolve the differences between the NMR-derived structural models. Potential binding sites for Psb27 within PSII are discussed in light of recent biochemical data in the literature.     

Crystal Structure of Monomeric Photosystem II from Thermosynechococcus elongatus at 3.6-? Resolution

The membrane-embedded photosystem II core complex (PSIIcc) uses light energy to oxidize water in photosynthesis. Information about the spatial structure of PSIIcc obtained from x-ray crystallography was so far derived from homodimeric PSIIcc of thermophilic cyanobacteria. Here, we report the first crystallization and structural analysis of the monomeric form of PSIIcc with high oxygen evolution capacity, isolated from Thermosynechococcus elongatus. The crystals belong to the space group C222₁, contain one monomer per asymmetric unit, and diffract to a resolution of 3.6 Å. The x-ray diffraction pattern of the PSIIcc-monomer crystals exhibit less anisotropy (dependence of resolution on crystal orientation) compared with crystals of dimeric PSIIcc, and the packing of the molecules within the unit cell is different. In the monomer, 19 protein subunits, 35 chlorophylls, two pheophytins, the non-heme iron, the primary plastoquinone Q_A, two heme groups, 11 β-carotenes, 22 lipids, seven detergent molecules, and the Mn₄Ca cluster of the water oxidizing complex could be assigned analogous to the dimer. Based on the new structural information, the roles of lipids and protein subunits in dimer formation of PSIIcc are discussed. Due to the lack of non-crystallographic symmetry and the orientation of the membrane normal of PSIIcc perpendicular (∼87°) to the crystallographic b-axis, further information about the structure of the Mn₄Ca cluster is expected to become available from orientation-dependent spectroscopy on this new crystal form.     

Expression of yeast acyl-CoA-∆9 desaturase leads to accumulation of unusual monounsaturated fatty acids in soybean seeds

An acyl-CoA-Δ9 desaturase from Saccharomyces cerevisiae was expressed by subcellular-targeting in soybean (Glycine max) seeds with the goal of increasing palmitoleic acid (16:1Δ9), a high-valued fatty acid (FA), and simultaneously decreasing saturated FA in oil. The expression resulted in the conversion of palmitic acid (16:0) to 16:1Δ9 in soybean seeds. 16:1Δ9 and its elongation product cis-vaccenic acid (18:1Δ11) were increased to 17 % of the total fatty acids by plastid-targeted expression of the enzyme. Other lipid changes include the decrease of polyunsaturated FA and saturated FA, suggesting that a mechanism exists downstream in oil biosynthesis to compensate the FA alternation. This is the first time a cytosolic acyl-CoA-?9 desaturase is functionally expressed in plastid and stronger activity was achieved than its cytosolic expression. The present study provides a new strategy for converting 16:0 to 16:1Δ9 by engineering acyl-CoA-Δ9 desaturase in commercialized oilseeds.     

Evidence for a role of raffinose in stabilizing photosystem II during freeze–thaw cycles

A role of non-reducing sugars like sucrose and raffinose in the protection of plant cells against damage during freezing has been proposed for many species, but reports on physiological effects are conflicting. Non-aqueous fractionation of mesophyll cell compartments in Arabidopsis thaliana was used to show that sucrose and raffinose accumulate in plastids during low temperatures, pointing to a physiological role in protecting the photosynthetic apparatus. Comparing a previously described raffinose synthase (RS) mutant of A. thaliana with its corresponding wild type, accession Col-0, revealed that a lack of raffinose has no effect on electrolyte leakage from leaf cells after freeze–thaw cycles, supporting that raffinose is not essential for protecting the plasma membrane. However, in situ chlorophyll fluorescence showed that maximum quantum yield of PS II photochemistry (F _v/F _m) and other fluorescence parameters of cold acclimated leaves subjected to freeze–thaw cycles were significantly lower in the raffinose synthase mutant than in the corresponding wild type, indicating that raffinose is involved in stabilizing PS II of cold acclimated leaf cells against damage during freezing.     

Disproportionation of pentaammineruthenium(III)–nucleoside complexes leads to two-electron oxidation of nucleosides without involving oxygen molecules

Pentaammineruthenium(III) complexes of deoxyinosine (dIno) and xanthosine (Xao) ([Ru^III(NH₃)₅(L)], L?is?dIno, Xao) in basic solution were studied by UV?Cvis spectroscopy, liquid chromatography/electrospray ionization mass spectrometry, and high-performance liquid chromatography. Both Ru^III complexes disproportionate to Ru^II and Ru^IV. Disproportionation followed the rate law d[Ru^II]/dt?=?(k _o?+?k ₁[OH^?])[Ru^III]. k _o and k ₁ of disproportionation at 25?°C were 2.1 (±0.1)?×?10^?3?s^?1 and 21.4?±?3.2?M^?1 s^?1, respectively, for [Ru^III(NH₃)₅(dIno)], and 3.5 (±0.7)?×?10^?4?s^?1 and 59.7?±?3.6?M^?1?s^?1, respectively, for [Ru^III(NH₃)₅(Xao)]. The [Ru^III(NH₃)₅(Xao)] complex disproportionates at a faster rate than [Ru^III(NH₃)₅(dIno)] owing to the stronger electron-withdrawing effect of exocyclic oxygen in Xao. The activation parameters ??H ^? and ??S ^? for k ₁ of [Ru^III(NH₃)₅(dIno)] were 80.2?±?15.2?kJ?mol^?1 and 47.6?±?9.8?J?K^?1 mol^?1, respectively, indicating that the disproportionation of Ru^III to Ru^II and Ru^IV is favored owing to the positive entropy of activation. The final products of both complexes in basic solution under Ar were compared with those under O₂. Under both conditions [Ru(NH₃)₅(8-oxo-L)] was produced, but via different mechanisms. In both aerobic and anaerobic conditions, the deprotonation of highly positively polarized C8-H of Ru-L by OH^? initiates a two-electron redox reaction. For the next step, we propose a one-step two-electron redox reaction between L and Ru^IV under anaerobic conditions, which differentiates from Clarke??s mechanism of two consecutive one-electron redox reactions between L, Ru^III, and O₂.     

Binary complexes of Mg(II) and Ca(II) in water–surfactant mixtures

Abstract

Chemical speciation of Mg(II) and Ca(II) complexes of L-histidine in the presence of water–surfactant mixtures in the concentration range 0.0–2.5% w/v CTAB and SDS, 0.0–5.0% v/v TX-100 maintaining an ionic strength of 0.16 mol dm^?3 at 303 K has been studied pH metrically. The active forms of the ligand are LH₃²⁺, LH₂⁺, LH and L^?. The models containing different numbers of species were refined by using the computer program, MINIQUAD75. The predominant species detected were ML₂H₄⁴⁺, ML₂H₃³⁺, ML₂H₂²⁺, and ML₂. The best fit chemical models were arrived at based on statistical parameters. The trend in variation of complex stability constants with change in the composition of the medium is explained on the basis of electrostatic and non-electrostatic forces. The effect of errors in the stability constants was also studied. Chemical speciation was also discussed based on the distribution diagrams.     

Variation in the methods leads to variation in the interpretation of biodiversity–ecosystem multifunctionality relationships

生物多样性常常和生态系统多功能性(生态系统同时提供多个生态系统功能的能力)正相关。然而,生物多样性与生态系统多功能性的关系是否依赖于生态系统功能的数目有诸多争议。其中,生物多样性对生态系统多功能性的影响或许不随生态系统功能数目的变化而变化,或者随生态系统功能数目的增多而增强。我们期望通过研究不同生态系统多功能性指数的统计原理来解决这些争议。 我们使用了模型模拟和一系列来自不同空间尺度(从局域到全球)和不同生物群系(温带和高寒草地、森林和干旱地)的经验数据。我们回顾了量化生态系统多功能性的三种方法,包括平均值法、加和法和阈值法。我们发现随着生态系统功能数目的增加,生物多样性与生态系统多功能性的关系要么不变,要么增强。这些结果可由平均和加和的多功能性指数的统计原理来解释。具体来讲,当利用生态系统功能的平均值计算多功能性指数时,由于多样性对多功能性的效应等于多样性对单个生态系统功能效应的平均值,所以不会随生态系统功能数目的变化而变化。同样的道理,当利用单个生态系统的加和值计算多功能性指数时,多样性的效应会随着生态系统功能数目的增加而增强。我们提出了一个改进的多功能性指数,将平均或加和多功能性指数转化为标准化的多功能性指数, 以便于对不同研究的结果进行比较。此外,我们提出了基于变量数值范围的标准化方法来解决阈值法的数学假象问题(多样性效应随生态系统功能数目的增加而增强)。我们的研究结果表明,量化多功能性指数的方法不同,结果也不同。因此,有必要加深对不同方法数理基础的理解。而标准化的多功能性指数为比较不同研究中的生物多样性与生态系统多功能性的关系提供了有效的方法。     

Inhibition of Calvin–Benson cycle suppresses the repair of photosystem II in Symbiodinium: implications for coral bleaching

Inhibition of Calvin–Benson cycle (CBC) activity by thermal stress has been hypothesized to cause photoinhibition of photosystem II (PSII) in zooxanthellae of reef-building corals and consequently lead to bleaching. This study tests whether the interruption of CBC by glycolaldehyde (GA) leads to photoinhibition and subsequent coral bleaching in Stylophora pistillata. When S. pistillata was incubated with GA, the O₂ evolution rate declined in a dose-dependent manner and the extent of photoinhibition, reflected by a decreased maximum quantum yield of PSII (F _v/F _m), was enhanced. The effect of GA on photoinhibition was similar to that of chloramphenicol (CAP), an inhibitor of protein synthesis in chloroplasts. When S. pistillata was incubated in weak light following a high-light-induced photoinhibitory treatment, the recovery of PSII from photoinhibition was suppressed in a similar manner to both GA- and CAP-treated samples. After incubation in moderate light at 26°C, S. pistillata showed a bleaching response only in presence of GA. These results suggest that coral bleaching-like responses are caused by interruption of the CBC activity in S. pistillata and are associated with accelerated photoinhibition through suppression of the protein synthesis-dependent repair of PSII but not to an increase in photodamage to PSII.     

Involvement of sulfoquinovosyl diacylglycerol in the structural integrity and heat-tolerance of photosystem II

To examine the role of sulfoquinovosyl diacylglycerol (SQDG) in thylakoid membranes, we compared the structural and functional properties of photosystem II (PSII) between a mutant of Chlamydomonas reinhardtii defective in SQDG ( hf-2) and the wild type. The PSII core complex of hf-2, as compared with that of the wild type, showed structural fragility when solubilized with a detergent, dodecyl beta- d-maltoside, suggesting that the physical properties of the PSII complex were altered by the loss of SQDG. On the other hand, exposure of the cells to 41 degrees C for 120 min in the dark decreased the PSII activity to 70% and 50% of the initial levels in the wild type and hf-2, respectively, which implies that the PSII activity, in the absence of SQDG, becomes less stable under heat-stress conditions. PSII inactivated to 60% of the initial level by dark incubation at 41 degrees C was reactivated by following illumination even at 41 degrees C to more than 90% in the wild type, but only to 70% in hf-2. These results suggest that PSII inactivated by heat recovers through some mechanism dependent on light, and that SQDG participates in functioning of the mechanism. The conformational disorder of PSII caused by the defect in SQDG might be correlated with the increased susceptibility of its activity to heat-stress.     

The acceptor side of photosystem II is damaged more severely than the donor side of photosystem II in ‘Honeycrisp’ apple leaves with zonal chlorosis

To investigate the photoinhibition of photosynthesis in ‘Honeycrisp’ apple (Malus domestica Borkh. cv. Gala) leaves with zonal chlorosis, we compared pigments, CO₂ assimilation and chlorophyll (Chl) a fluorescence (OJIP) transient between chlorotic leaves and normal ones. Chl and carotenoids (Car) contents, Chl a/b ratio, and absorptance were lower in chlorotic leaves than in normal ones, whereas Car/Chl ratio was higher in the former. Although CO₂ assimilation and stomatal conductance were lower in chlorotic leaves, intercellular CO₂ concentration did not differ significantly between the two leaf types. Compared with normal leaves, chlorotic ones had increased deactivation of oxygen-evolving complexes (OEC), minimum fluorescence (F _o), dissipated energy, relative variable fluorescence at L-, W-, J- and I-steps, and decreased maximum fluorescence (F _m), maximum quantum yield for primary photochemistry (F _v /F _m or TR_o/ABS), quantum yield for electron transport (ET_o/ABS), quantum yield for the reduction of end acceptors of photosystem I (PSI) (φ_Ro and RE_o/ABS), maximum amplitude of IP phase, amount of active photosystem II (PSII) reaction centers (RCs) per cross section (CS) and total performance index (PI_tot,abs). In conclusion, photoinhibition occurs at both the donor (i.e., the OEC) and the acceptor sides of PSII in chlorotic leaves. The acceptor side is damaged more severely than the donor side, which possibly is the consequence of over-reduction of PSII due to the slowdown of Calvin cycle. In addition to decreasing light absorptance by lowering Chl level, energy dissipation is enhanced to protect chlorotic leaves from photo-oxidative damage.     

Speciation studies of Co(II), Ni(II) and Cu(II) complexes of L-valine in acetonitrile–water mixtures

Abstract

Chemical speciation of binary complexes of Co(II), Ni(II) and Cu(II) with L-valine in 0.0-60.0% v/v acetonitrile-water mixtures was studied pH-metrically at an ionic strength of 0.16 mol L^-1 at 303.0 K. The existence of different binary complexes was established from the modelling studies, using the computer program MINIQUAD75. The appropriateness of the model was ascertained by studying the effect of errors in concentrations of the reagents. The trend in variation of stability constants with change in the permittivity of the medium is explained on the basis of electrostatic and non-electrostatic forces. The species distribution diagrams and the plausible equilibria for the formation of the species are also presented.