High-light induced superoxide radical formation in cytochrome b6f complex from Bryopsis corticulans as detected by EPR spectroscopy.

The generation of superoxide radical (O2·â») in Cyt b6f of Bryopsis corticulans under high light illumination was studied using electron paramagnetic resonance (EPR) spectroscopy. This could be evidenced by the addition of SOD which specifically reacted with O2·â». The generation of O2·â» was lost in the absence of oxygen and was found to be suppressed in the presence of NaN3 and be scavenged by extraneous antioxidants such as ascorbate, ß-carotene and glutathione which could also scavenged ¹O2*. These results indicated that O2·â» which produced under high light illumination in Cyt b6f of B. corticulans might rise from a reaction which ¹O2* could participated in. Also the photo-protection mechanism to Cyt b6f complex by antioxidants which might contain in thylakoid was speculated.

Proteomic analysis of hydrogen photoproduction in sulfur-deprived Chlamydomonas cells.

The green alga Chlamydomonas reinhardtii is a model organism to study H(2) metabolism in photosynthetic eukaryotes. To understand the molecular mechanism of H(2) metabolism, we used 2-DE coupled with MALDI-TOF and MALDI-TOF/TOF-MS to investigate proteomic changes of Chlamydomonas cells that undergo sulfur-depleted H(2) photoproduction process. In this report, we obtained 2-D PAGE soluble protein profiles of Chlamydomonas at three time points representing different phases leading to H(2) production. We found over 105 Coomassie-stained protein spots, corresponding to 82 unique gene products, changed in abundance throughout the process. Major changes included photosynthetic machinery, protein biosynthetic apparatus, molecular chaperones, and 20S proteasomal components. A number of proteins related to sulfate, nitrogen and acetate assimilation, and antioxidative reactions were also changed significantly. Other proteins showing alteration during the sulfur-depleted H(2) photoproduction process were proteins involved in cell wall and flagella metabolisms. In addition, among these differentially expressed proteins, 11 were found to be predicted proteins without functional annotation in the Chlamydomonas genome database. The results of this proteomic analysis provide new insight into molecular basis of H(2) photoproduction in Chlamydomonas under sulfur depletion.

High-light induced singlet oxygen formation in cytochrome b(6)f complex from Bryopsis corticulans as detected by EPR spectroscopy.

Electron paramagnetic resonance (EPR) spectroscopy was used to detect the light-induced formation of singlet oxygen ((1)O(2)*) in the intact and the Rieske-depleted cytochrome b(6)f complexes (Cyt b(6)f) from Bryopsis corticulans, as well as in the isolated Rieske Fe-S protein. It is shown that, under white-light illumination and aerobic conditions, chlorophyll a (Chl a) bound in the intact Cyt b(6)f can be bleached by light-induced (1)O(2)*, and that the (1)O(2)* production can be promoted by D(2)O or scavenged by extraneous antioxidants such as l-histidine, ascorbate, beta-carotene and glutathione. Under similar experimental conditions, (1)O(2)* was also detected in the Rieske-depleted Cyt b(6)f complex, but not in the isolated Rieske Fe-S protein. The results prove that Chl a cofactor, rather than Rieske Fe-S protein, is the specific site of (1)O(2)* formation, a conclusion which draws further support from the generation of (1)O(2)* with selective excitation of Chl a using monocolor red light.

Singlet oxygen formation and chlorophyll a triplet excited state deactivation in the cytochrome b6f complex from Bryopsis corticulans.

We have attempted to investigate the correlation between the detergent-perturbed structural integrity of the Cyt b (6) f complex from the marine green alga Bryopsis corticulans and its photo-protective properties, for which the nonionic detergents n-octyl-beta-D-glucopyranoside (beta-OG) and n-dodecyl-beta-D-maltoside (beta-DM), respectively, were used for the preparation of Cyt b (6) f, and the singlet oxygen ((1)O(2)*) production as well as the triplet excited-state chlorophyll a ((3)Chl a*) formation and deactivation were examined by spectroscopic means. Near-infrared luminescence of (1)O(2)* (approximately 1,270 nm) on photo-irradiation was detected for the beta-OG preparation where the complex is mainly in oligomeric state, but not for the beta-DM one in which the complex exists in dimeric form. Under anaerobic condition, photo-excitation of Chl a in the beta-DM preparation generated (3)Chl a* with a lower quantum yield of Phi(T) approximately 0.02 and a longer lifetime of approximately 600 micros with respect to those as in the case of beta-OG preparation, Phi(T) approximately 0.12 and 200-300 micros. These results prove that the enzymatically active and intact Cyt b (6) f complex on photo-excitation tends to produce little (3)Chl a* or (1)O(2)*, which implies that the pigment-protein assembly of Cyt b (6) f complex per se is crucial for photo-protection.

Proteomic analysis of plasma membranes of cyanobacterium Synechocystis sp. Strain PCC 6803 in response to high pH stress.

Cyanobacteria are unique prokaryotes possessing plasma-, outer- and thylakoid membranes. The plasma membrane of a cyanobacterial cell serves as a crucial barrier against its environment and is essential for biogenesis of cyanobacterial photosystems. Previously, we have identified 79 different proteins in the plasma membrane of Synechocystis sp. Strain PCC 6803 based on 2D- and 1D- gels and MALDI-TOF MS. In this work, we have performed a proteomic study screening for high-pH-stress proteins in Synechocystis. 2-D gel profiles of plasma membranes isolated from both control and high pH-treated cells were constructed and compared quantitatively based on different protein staining methods including DIGE analysis. A total of 55 differentially expressed protein spots were identified using MALDI-TOF MS and MALDI-TOF/TOF MS, corresponding to 39 gene products. Twenty-five proteins were enhanced/induced and 14 reduced by high pH. One-third of the enhanced/induced proteins were transport and binding proteins of ABC transporters including 3 phosphate transport proteins. Other proteins include MinD involved in cell division, Cya2 in signaling and proteins involved in photosynthesis and respiration. Furthermore, among these proteins regulated by high pH, eight were found to be hypothetical proteins. Functional significance of the high-pH-stress proteins is discussed integrating current knowledge on cyanobacterial cell physiology.

Exploring the mechanism of Physcomitrella patens desiccation tolerance through a proteomic strategy.

The moss Physcomitrella patens has been shown to tolerate abiotic stresses, including salinity, cold, and desiccation. To better understand this plant's mechanism of desiccation tolerance, we have applied cellular and proteomic analyses. Gametophores were desiccated over 1 month to 10% of their original fresh weight. We report that during the course of dehydration, several related processes are set in motion: plasmolysis, chloroplast remodeling, and microtubule depolymerization. Despite the severe desiccation, the membrane system maintains integrity. Through two-dimensional gel electrophoresis and image analysis, we identified 71 proteins as desiccation responsive. Following identification and functional categorization, we found that a majority of the desiccation-responsive proteins were involved in metabolism, cytoskeleton, defense, and signaling. Degradation of cytoskeletal proteins might result in cytoskeletal disassembly and consequent changes in the cell structure. Late embryogenesis abundant proteins and reactive oxygen species-scavenging enzymes are both prominently induced, and they might help to diminish the damage brought by desiccation.

WITHDRAWN: Effect of monogalactosyldiacylglycerol on the interactions between light-harvesting complexes II and photosystem II core complexes reconstituted into liposomes constituted of thylakoid lipids.

This article has been withdrawn consistent with Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy). The Publisher apologizes for any inconvenience this may cause.

Study on energy transfer between carotenoid and chlorophyll a in cytochrome b6f complex from Bryopsis corticulans.

The excitation energy transfer between carotenoid and chlorophyll (Chl) in the cytochrome b ( 6 ) f complex from Bryopsis corticulans (B. corticulans), in which the carotenoid is 9-cis-alpha-carotene, was investigated by means of fluorescence excitation and sub-microsecond time-resolved absorption spectroscopies. The presence of efficient singlet excitation transfer from alpha-carotene to Chl a was found with an overall efficiency as high as approximately approximately 24%, meanwhile the Chl a-to-alpha-carotene triplet excitation transfer was also evidenced. Circular dichroism spectroscopy showed that alpha-carotene molecule existed in an asymmetric environment and Chl a molecule had a certain orientation in this complex.

Steady state fluorescence spectroscopy of the photosystem II core complex.

Spectroscopic properties within the core complex of photosystem II were investigated by studying the influence of the wavelength of excitation on the fluorescence emission spectrum. At two temperatures, when the core complex of PSII isolated from spinach was excited at six different excitation wavelengths ranging from 436 nm to 520 nm, there is no difference in the maxima of the emission spectra of the core complex, and when the core complex was excited at 480, 489, 495 and 507 nm respectively, fluorescence intensities of maxima decrease with increasing of the absorbance of the beta-carotene molecules at the four excitation wavelengths. The extent of change of the shoulder of the spectra beyond 700 nm depends on the kind of pigment molecule excited. The excitation wavelength can influence the way of energy transfer in the core complex of photosystem II. By Gaussian deconvolution analysis, at least seven groups of chlorophyll a molecules were discovered. They are Chl a(660), Chl a(670), Chl a(680), Chl a(682), Chl a(684), Chl a(687) and Chl a(690).

Ultrafast carotenoid-to-chlorophyll singlet energy transfer in the cytochrome b6f complex from Bryopsis corticulans.

Ultrafast carotenoid-to-chlorophyll (Car-to-Chl) singlet excitation energy transfer in the cytochrome b(6)f (Cyt b(6)f) complex from Bryopsis corticulans is investigated by the use of femtosecond time-resolved absorption spectroscopy. For all-trans-alpha-carotene free in n-hexane, the lifetimes of the two low-lying singlet excited states, S(1)(2A(g)(-)) and S(2)(1B(u)(+)), are determined to be 14.3 +/- 0.4 ps and 230 +/- 10 fs, respectively. For the Cyt b(6)f complex, to which 9-cis-alpha-carotene is bound, the lifetime of the S(1)(2A(g)(-)) state remains unchanged, whereas that of the S(2)(1B(u)(+)) state is significantly reduced. In addition, a decay-to-rise correlation between the excited-state dynamics of alpha-carotene and Chl a is clearly observed. This spectroscopic evidence proves that the S(2)(1B(u)(+)) state is able to transfer electronic excitations to the Q(x) state of Chl a, whereas the S(1)(2A(g)(-)) state remains inactive. The time constant and the partial efficiency of the energy transfer are determined to be 240 +/- 40 fs and (49 +/- 4)%, respectively, which supports the overall efficiency of 24% determined with steady-state fluorescence spectroscopy. A scheme of the alpha-carotene-to-Chl a singlet energy transfer is proposed based on the excited-state dynamics of the pigments.

Hydrogen peroxide-induced chlorophyll a bleaching in the cytochrome b6f complex: a simple and effective assay for stability of the complex in detergent solutions.

The instability of cytochrome b ( 6 ) f complex in detergent solutions is a well-known problem that has been studied extensively, but without finding a satisfactory solution. One of the important reasons can be short of the useful method to verify whether the complex suspended in different detergent is in an intact state or not. In this article, a simple and effective assay for stability of the complex was proposed based on the investigation on the different effects of the two detergents, n-octyl-beta-D: -glucopyranoside (OG) and dodecyl-beta-D: -maltoside (DDM), on the properties of the complex. DDM stabilizes the complex preparation more effectively whereas OG denatures the interactions of the heme groups and pigment molecules with the protein environment, leading to the bleaching of chlorophyll a induced by addition of hydrogen peroxide. The assay of the use of hydrogen peroxide to characterize the complex by studying the bleaching of chlorophyll induced by hydrogen peroxide and the peroxidase activity of the complex was discussed. This simple method will probably be useful to study the stability of the complex.

Three-dimensional architecture of phycobilisomes from Nostoc flagelliforme revealed by single particle electron microscopy.

Phycobilisomes are protein complexes that harvest light and transfer energy to the photo system. Here, the three dimensional structure of intact phycobilisomes from Nostoc flagelliforme is studied by a combination of negative stain electron microscopy and cryo-electron microscopy. Results show that the intact phycobilisomes are composed of a tricylindrical core and six rods. Each allophycocyanin cylinder presents a double-layered structure when viewed from the side and a triangular shape when viewed from the top. These characteristics indicate that allophycocyanin trimers in the intact phycobilisomes are arranged into hexameric oligomers in a parallel manner.

[Effects of Triton X-100 on the oxygen uptake rate of photosystem I particles treated at 70 degrees C].

The characteristics including oxygen uptake rates, fluorescence spectra and absorption spectra of photosystem I particles with or without Triton-X 100 treatment before or after the incubation at 70 degrees C for 10 min were compared. The oxygen uptake rates of photosystem I particles decreased after being incubated at 70 degrees C for 10 min, which could be recovered by the addition of Triton-X 100. Singlet oxygen was formed when the light-harvesting complex I was separated from the core complex of photosystem I, which resulted in high oxygen uptake rate. There was much difference in the fluorescence spectra of photosystem I particles between photosystem I particles treated with Triton-X 100 after the incubation at 70 degrees C for 10 min or not, which implies the ability of Triton-X 100 to promote the recovery of photosystem I particles after the incubation at 70 degrees C for 10 min.

Effect of various temperatures on phosphatidylglycerol biosynthesis in thylakoid membranes.

Lipid unsaturation, the major factor to maintain thylakoid membrane fluidity, is affected by temperature. In this work, we analysed the molecular species composition of phosphatidylglycerol (PG) in thylakoid membranes during spinach (Spinacia oleracea) and squash (Cucurbita pepo) cotyledon growth to investigate how the growth temperature affects the PG biosynthesis. Of the 10 molecular species detected, temperature affected mainly the relative content of molecular species containing linolenic acid (18:3) and those containing palmitic acid (16:0) at the sn-1 position of glycerol backbone. Lowering the temperature induced an increase in the former and a decrease in the latter. The relative content of molecular species containing 18:3 or 16:0 at the sn-1 position of the glycerol backbone were correlated with temperature. Our results indicate that the substrate selectivity of the glycerol-3-phosphate acyltransferase (GPAT) in chloroplasts towards 16:0 or oleic acid (18:1) and the activity of fatty acid desaturases are greatly affected by temperature. In addition, changes in the relative content of PG molecular species induced by variations in growth temperature depended mainly on the substrate selectivity of GPAT.

Photosynthesis research in the People's Republic of China.

The first research paper on photosynthesis in China was published by T.T. Li(2) in 1929. Two photosynthesis laboratories were established in Shanghai and Beijing in the 1950s and the 1960s, respectively. A photophosphorylation 'intermediate' was discovered after the energy conversion process was separated into light and dark phases in the 1960s. Since the 1980s, research has accelerated at several different levels through efforts of a large number of scientists in China.