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1.
Plastocyanin as the possible site of photosynthetic electron transport inhibition by glutaraldehyde 总被引:3,自引:2,他引:1
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Treatment of spinach chloroplasts with glutaraldehyde causes an inhibition in the electron transport chain between the two photosystems. Measurements of O2 flash yields, pH exchange, and fluorescence induction show that the O2 evolving apparatus, photosystem II and its electron acceptor pool are not affected. The behavior of P700 indicates that its reduction but not its oxidation, is severely inhibited. Cytochrome f is still reducible by photosystem II but also slowly oxidizable by photosystem I. The sensitivity of isolated plastocyanin to glutaraldehyde further supports the conclusion that glutaraldehyde inhibits at the plastocyanin level and thereby induces a break between P700 and cytochrome f. 相似文献
2.
Role of bicarbonate in photosystem II, the water-plastoquinone oxido-reductase of plant photosynthesis 总被引:4,自引:0,他引:4
Depletion of bicarbonate (carbon dioxide) from oxygenic cells or organelles not only causes cessation of carbon dioxide fixation, but also a strong decrease in the activity of photosystem II; the photosystem II activity can be restored by readdition of bicarbonate. Effects of bicarbonate exist on both the acceptor as well as on the donor side of photosystem II. The influence on the acceptor side is located between the primary and secondary quinone electron acceptor of photosystem II, and can be demonstrated in intact cells or leaves as well as in isolated thylakoids and reaction center preparations. At physiological pH, bicarbonate ions are suggested to form hydrogen bonds to several amino acids on both D1 and D2 proteins, the reaction center subunits of photosystem II, as well as to form ligands to the non-heme iron between the D1 and D2 proteins. Bicarbonate, at physiological pH, has an important role in the water-plastoquinone oxido-reductase: on the one hand it may stabilize, by conformational means, the reaction center protein of photosystem II that allows efficient electron flow and protonation of certain amino acids near the secondary quinone electron acceptor of photosystem II; and, on the other hand, it akppears to play a significant role in the assembly or functioning of the manganese complex at the donor side. Functional roles of bicarbonate in vivo, including protection against photoinhibition, are also discussed. 相似文献
3.
《Archives of biochemistry and biophysics》1985,241(2):540-549
The concentration of photosystem II was determined in thylakoids of dwarf peas by the use of the following methods: absorption change at 325 nm; atrazine binding; and flash yields of oxygen evolution (Emerson-Arnold method), of protons from oxidation of water, and of reduction of DCIP. For the first time all of the flash-yield measurements have been done on the same sample and give equivalent values for the concentration of photosystem II. Agreement of the absorption change measurement at 325 nm with the other measurements was accomplished by the introduction of important improvements to the methods of Melis and co-workers [Proc. Natl. Acad. Sci. USA (1980)77, 4712–4716]. The atrazine-binding method gave photosystem II values that were twice as large as any of the other photosystem II measurements. Possible reasons are discussed for this discrepancy in terms of the secondary acceptor (Q400) of Ikegami and Katoh [Plant Cell. Physiol. (1973)14, 829–836]. The concentration of photosystem I was measured by absorption change at 705 nm. From the concentration values of photosystem II and I the system II/I stoichiometry was calculated. 相似文献
4.
The triplet states of photosystem II core particles from spinach were studied using time-resolved cw EPR technique at different reduction states of the iron-quinone complex of the reaction center primary electron acceptor. With doubly reduced primary acceptor, the well-known photosystem II triplet state characterised by zero-field splitting parameters |D| = 0.0286 cm−1, |E| = 0.0044 cm−1 was detected. When the primary acceptor was singly reduced either chemically or photochemically, a triplet state of a different spectral shape was observed, bearing the same D and E values and characteristic spin polarization pattern arising from RC radical pair recombination. The latter triplet state was strongly temperature dependent disappearing at T = 100 K, and had a much faster decay than the former one. Based on its properties, this triplet state was also ascribed to the photosystem II reaction center. A sequence of electron-transfer events in the reaction centers is proposed that explains the dependence of the triplet state properties on the reduction state of the iron-quinone primary acceptor complex. 相似文献
5.
Photosystem II Core Phosphorylation Heterogeneity, Differential Herbicide Binding, and Regulation of Electron Transfer in Photosystem II Preparations from Spinach 总被引:1,自引:0,他引:1
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The effect of photosystem II core phosphorylation on the secondary quinone acceptor of photosystem II (QB) domain environment was analyzed by comparative herbicide-binding studies with photosystem II preparations from spinach (Spinacia oleracea L.). It was found that phosphorylation reduces the binding affinity for most photosynthetic herbicides. The binding of synthetic quinones and of the electron acceptor 2,6-dichlorophenolindophenol is also reduced by photosystem II phosphorylation. Four photosystem II core populations isolated from membranes showed different extents of phosphorylation as well as different degrees of affinity for photosynthetic herbicides. These findings support the idea that heterogeneity of photosystem II observed in vivo could be, in part, due to phosphorylation. 相似文献
6.
Two sites in the photosynthetic electron transport chain of spinach chloroplasts are sensitive to inhibition by the plastoquinone antagonist dibromothymoquinone (2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone). This compound imposes maximal inhibition on reactions involving electron transport from water to a terminal acceptor such as ferricyanide at concentrations of about 1 μm. At concentrations of about 10 μm, dibromothymoquinone also inhibits electron transport reactions catalyzed by photosystem II in the presence of p-phenylenediimines or p-benzoquinones. This inhibition is observed in both untreated and -inhibited chloroplast preparations. Thiol incubation of chloroplasts exposed to dibromothymoquinone relieves inhibition at both sites. This reversal of inhibition is, however, different for the two sites. Restoration of ferricyanide reduction, which is blocked by 1 μm dibromothymoquinone, required high thiol/inhibitor ratios and incubation times with thiol of up to 3 min. The reversal of inhibition of p-phenylenediimine reduction by photosystem II, on the other hand, requires a thiol/inhibitor ratio of 1, and incubation times as short as 5 s. Addition of bovine serum albumin to absorb dibromothymoquinone results in a partial restoration of photosystem II reactions, but ferricyanide reduction, which requires photosystem II and photosystem I, cannot be restored by this procedure. 相似文献
7.
Robert L. Heath 《Biophysical journal》1970,10(12):1173-1188
The chlorophyll fluorescence yield in isolated chloroplasts without an added electron acceptor is increased by actinic illumination. The decline in the fluorescence yield when the actinic illumination is extinguished can be accurately represented by three, independent, exponential decays with half-times of approximately 0.8, 5, and 30 sec. These results have been interpreted using Duysens' theory of fluorescence quenching by a compound (Q) on the reducing side of photosystem II. This theory states that changes in fluorescence yield are indicative of electron flow through Q. The most rapid decay is eliminated by an EDTA washing of the chloroplasts and the half-time is increased by uncoupling with ammonia and by added electron acceptors in suboptimal concentrations. Thus, this decay may represent electron flow from Q to intermediates on the oxidizing side of photosystem I. The decay with a half-time of 5 sec is affected in the same manner as the decay with the shortest half-time by the same procedures. However, electron donors to photosystem II lengthen the half-time of the 5 sec decay while eliminating the most rapid decay. This 5 sec decay can be interpreted as electron flow from Q to intermediates either on the reducing side of photosystem II or on the oxidizing side of photosystem I. The decay with the longest half-time is affected only by pH and electron donors to photosystem II. Therefore, this decay may indicate electron flow from Q to intermediates on the oxidizing side of photosystem II which may be connected to the regeneration of the oxygen burst. 相似文献
8.
Characterization of a Purified Photosystem II-Phycobilisome Particle Preparation from Porphyridium cruentum 总被引:2,自引:1,他引:1
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Detergent preparations isolated from thylakoids of the red alga Porphyridium cruentum, in a sucrose, phosphate, citrate, magnesium chloride medium consist of phycobilisomes and possess high rates of photosystem II activity. Characterization of these particles shows that the O2-evolving activity is stable for several hours and the pH optimum is about 6.5 to 7.2. Response of the system to light, electron donors and acceptors, and inhibitors verify that the observed activity, measured both as O2 evolution and 2,6-dichlorophenol-indophenol reduction, is due to photosystem II. Furthermore, photosystem II is functionally coupled to the phycobilisome in this preparation since green light, absorbed by phycobilisomes of P. cruentum, is effective in promoting both O2 evolution and 2,6-dichlorophenol-indophenol reduction. Photosystem II activity declines when light with wavelengths shorter than 665 nm is removed. Both 3-(3,4-dichlorophenyl)-1,1-dimethylurea and atrazine inhibit photosystem II activity in this preparation, indicating that the herbicide binding site is a component of the photosystem II-phycobilisome particle. 相似文献
9.
Comparison of Photosynthetic Performance in Triazine-Resistant and Susceptible Biotypes of Amaranthus hybridus 总被引:6,自引:5,他引:1
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The rate of CO2 reduction in the S-triazine-resistant biotype of smooth pigweed (Amaranthus hybridus L.) was lower at all levels of irradiance than the rate of CO2 reduction in the susceptible biotype. The intent of this study was to determine whether or not the lower rates of CO2 reduction are a direct consequence of the same factors which confer triazine resistance. The quantum yield of CO2 reduction was 23 ± 2% lower in the resistant biotype of pigweed and the resistant biotype of pigweed had about 25% fewer active photosystem II centers on both a chlorophyll and leaf area basis. This quantum inefficiency of the resistant biotype can be accounted for by a decrease in the equilibrium constant between the primary and secondary quinone acceptors of the photosystem II reaction centers which in turn would lead to a higher average level of reduced primary quinone acceptor in the resistant biotype. Thus, the photosystem II quantum inefficiency of the resistant biotype appears to be a direct consequence of those factors responsible for triazine resistance but a caveat to this conclusion is discussed. The effects of the quantum inefficiency of photosystem II on CO2 reduction should be overcome at high light and therefore cannot account for the lower light-saturated rate of CO2 reduction in the resistant biotype. Chloroplast lamellar membranes isolated from both triazine-resistant and triazine-susceptible pigweed support equivalent rates of whole chain electron transfer and these rates are sufficient to account for the rate of light-saturated CO2 reduction. This observation shows that the slower transfer of electrons from the primary to the secondary quinone acceptor of photosystem II, a trait which is characteristic of the resistant biotype, is nevertheless still more rapid than subsequent reactions of photosynthetic CO2 reduction. Thus, it appears that the lower rate of light-saturated CO2 reduction of the resistant biotype is not limited by electron transfer capacity and therefore is not a direct consequence of those factors which confer triazine resistance. 相似文献
10.
D B Knaff 《Biochimica et biophysica acta》1975,376(3):583-587
The primary electron acceptor of Photosystem II has a midpoint oxidation-reduction potential of +95 mV at pH 7.0 in Photosystem II chloroplast fragments prepared by digitonin treatment. The midpoint potential of the acceptor has a pH dependence of -60 mV/pH unit. At concentrations that inhibit oxygen evolution, o-phenanthroline shifts the midpoint potential of the primary acceptor by +70 mV. The shifted potential retains the same dependence on pH. The effect of o-phenanthroline suggests that it interacts directly with the primary electron acceptor of photosystem II in a manner similar to that reported previously for the primary electron acceptor in purple photosynthetic bacteria. 相似文献
11.
Baker NR 《Plant physiology》1978,62(6):889-893
The effects of wide concentration ranges of NaCl, KCl, and MgCl2 on ferricyanide reduction and the fluorescence induction curve of isolated spinach (Spinacia oleracea) chloroplasts were investigated. Concentrations of the monovalent salts above 100 mm and MgCl2 above 25 mm produced a decrease in the rate of ferricyanide reduction by thylakoids uncoupled with 2.5 mm NH4Cl which cannot be attributed to changes in the primary photochemical capacity of photosystem II. Salt-induced decreases in the effective concentration of the secondary electron acceptor of photosystem II, plastoquinone, reduce the capacity for secondary photochemistry of photosystem II and this could contribute to the reduction in ferricyanide reduction by uncoupled thylakoids at high salinities. The rate of ferricyanide reduction by coupled thylakoids is little affected by salinity changes, indicating that the rate-limiting phosphorylation mechanism in electron flow from water to ferricyanide in coupled thylakoids is salt-tolerant, whereas the rate-limiting reaction in uncoupled ferricyanide reduction is considerably affected by salinity changes. Salt-induced changes in the fluorescence induction curve are interpreted in terms of changes in the rate constants for excitation decay by radiationless transitions, exciton transfer from photosystem II chlorophylls to other associated chlorophyll species, and photochemistry. 相似文献
12.
Janet Banaszak Rita Barr Frederick L. Crane 《Journal of bioenergetics and biomembranes》1976,8(2):83-92
Various sites of ferricyanide reduction were studied in spinach chloroplasts. It was found that in the presence of dibromothymoquinone a fraction of ferricyanide reduction was dibromothymoquinone sensitive, implying that ferricyanide can be reduced by photosystem I as well as photosystem II. To separate ferricyanide reduction sites in photosystem II, orthophenanthroline and dichlorophenyl dimethylurea inhibitions were compared at various pH's. It was noted that at low pH ferricyanide reduction was not completely inhibited by orthophenanthroline. At high pH's, however, inhibition of ferricyanide reduction by orthophenanthroline was complete. It was found that varying concentration of orthophenanthroline at a constant pH showed different degrees of inhibition. In the study of ferricyanide reduction by photosystem II various treatments affecting plastocyanin were performed. It was found that Tween-20 or KCN treatments which inactivated plastocyanin did not completely inactivate ferricyanide reduction. These data support the conclusion that ferricyanide accepts electrons both before and after plastoquinone in photosystem II.Abbreviations DCMU
3-(3,4-dichlorophenyl)-1,1-dimethyurea
- MV
methyl viologen
- DBMIB
2,5-dibromothymoquinone
- DMBQ
2,6-dimethyl benzoquinone
- OP
1,10-orthophenanthroline
- TMPD
tetramethyl-p-phenylenediamine
- PS 1
photosystem I
- PS II
photosystem II
- SN
sucrose-sodium chloride chloroplasts
Supported by NSF Grant BMS 74-19689. 相似文献
13.
M G Goldfield R I Halilov S V Hangulov 《Biochemical and biophysical research communications》1978,85(3):1199-1203
The light-induced dark-reversible ESR signal in chloroplast fragments enriched in photosystem II and free from P700 contamination has been observed in the presence of silicomolybdate as an electron acceptor operating directly on the photosystem II primary acceptor. The signal at g = 2.0025 and with line-width ΔHpp = 9G rises and decays in close correlation with the photobleaching band centered at 680 nm and the minor peak at 435 nm. 相似文献
14.
p-Nitroacetophenoxime N-methylcarbamate (MCPNA) is a rather potent inhibitor of the electron transfer in spinach class A chloroplasts. In isolated thylakoids, MCPNA is an electron acceptor at the level of photosystem I (PS I). It inhibits O2 evolution in the presence of NADP and ferredoxin but not the reduction of ferricyanide. MCPNA is active as an acceptor between 3 μM and 100 μM. At concentrations higher than 300 μM, inhibition of photosystem II (PS II) occurs. MCPNA has no uncoupling effect on photophosphorylation. Reduction of MCPNA by thylakoids in the presence of light is in accordance with the E′o of this compound (??0.57 V) and is followed by an electron transfer to O2. This reaction probably explains the inhibitory effect of MCPNA on class A chloroplasts. 相似文献
15.
In this study, we evaluated how cadmium inhibitory effect on photosystem II and I electron transport may affect light energy
conversion into electron transport by photosystem II. To induce cadmium effect on the photosynthetic apparatus, we exposed
Chlamydomonas reinhardtii 24 h to 0–4.62 μM Cd2+. By evaluating the half time of fluorescence transients O–J–I–P at different temperatures (20–30°C), we were able to determine
the photosystem II apparent activation energies for different reduction steps of photosystem II, indicated by the O–J–I–P
fluorescence transients. The decrease of the apparent activation energies for PSII electron transport was found to be strongly
related to the cadmium-induced inhibition of photosynthetic electron transport. We found a strong correlation between the
photosystem II apparent activation energies and photosystem II oxygen evolution rate and photosystem I activity. Different
levels of cadmium inhibition at photosystem II water-splitting system and photosystem I activity showed that photosystem II
apparent activation energies are strongly dependent to photosystem II donor and acceptor sides. Therefore, the oxido-reduction
state of whole photosystem II and I electron transport chain affects the conversion of light energy from antenna complex to
photosystem II electron transport. 相似文献
16.
Oxygen consumption in photosystem II (PSII) preparations in the light was 2 mol O2/h per mg Chl at weakly acidic and at neutral pH values. It increased fourfold to fivefold at pH 8.5-9.0. The addition of either artificial electron donors for PSII such as MnCl2 or diphenylcarbazide, or diuron as an inhibitor of electron transfer from QA, the primary bound quinone acceptor, to QB, the secondary bound quinone acceptor of PSII, resulted in a decrease in oxygen consumption rate at basic pH to value close to ones measured at pH 6.5. Such additions did not affect oxygen consumption at lower pH values. The induction of variable chlorophyll fluorescence yield in the light differed greatly at pH 6.5 and 8.5. While at pH 6.5 the fluorescence yield, after an initial fast rise almost to Fmax, only slightly decreased, at pH 8.5 after such a rise it dropped promptly to a low value. The additions of the artificial electron donors at pH 8.5 resulted in the induction kinetics close to that observed at pH 6.5. These data indicate impairment of electron donation to P680+ that could be caused by damage to the water oxidation system at basic pH values. In experiments with PSII preparations treated with Tris to destroy the water-oxidizing complex, photoconsumption of oxygen in the entire pH region was close to the values in untreated preparations at basic pH. In untreated preparations the rate of light-induced oxygen consumption decreased in the presence of catalase, which decomposes H2O2, as well as in the presence of electron acceptor potassium ferricyanide. From these data it is suggested that the light-induced oxygen consumption in PSII is caused by two processes, by an interaction of O2 with organic radicals, which were formed due to oxidation of components of the donor side of this photosystem (proteins, lipids, pigments) by cation-radical P680+, as well as by oxygen reduction by still unidentified components of PSII. 相似文献
17.
Various sites of ferricyanide reduction were studied in spinach chloroplasts. It was found that in the presence of dibromothymoquinone a fraction of ferricyanide reduction was dibromothymoquinone sensitive, implying that ferricyanide can be reduced by photosystem I as well as photosystem II. To separate ferricyanide reduction sites in photosystem II, orthophenanthroline and dichlorophenyl dimethylurea inhibitions were compared at various pHs. It was noted that at low pH ferricyanide reduction was not completely inhibited by orothophenanthroline. At high pH's, however, inhibition of ferricyanide reduction by orthophenanthroline was complete. It was found that varying concentration of orthophenanthroline at a constant pH showed different degrees of inhibition. In the study of ferricyanide reduction by photosystem II various treatments affecting plastocyanin were performed. It was found that Tween-20 or KCN treatments which inactivated plastocyanin did not completely inactivate ferricyanide reduction. These data support the conclusion that ferricyanide accepts electrons both before and after plastoquinone in photosystem II. 相似文献
18.
The triplet states of photosystem II core particles from spinach were studied using time-resolved cw EPR technique at different reduction states of the iron--quinone complex of the reaction center primary electron acceptor. With doubly reduced primary acceptor, the well-known photosystem II triplet state characterised by zero-field splitting parameters |D|=0.0286 cm(-1), |E|=0.0044 cm(-1) was detected. When the primary acceptor was singly reduced either chemically or photochemically, a triplet state of a different spectral shape was observed, bearing the same D and E values and characteristic spin polarization pattern arising from RC radical pair recombination. The latter triplet state was strongly temperature dependent disappearing at T=100 K, and had a much faster decay than the former one. Based on its properties, this triplet state was also ascribed to the photosystem II reaction center. A sequence of electron-transfer events in the reaction centers is proposed that explains the dependence of the triplet state properties on the reduction state of the iron--quinone primary acceptor complex. 相似文献
19.
The effect of thylakoid phosphorylation on noncyclic electron transport in spinach chloroplasts was investigated by measuring both the reduction of nicotinamide adenine dinucleotide phosphate (NADP) and the steady-state redox level of the primary electron acceptor quinone of photosystem II (Q) during electron flow to NADP. These data are compared with the theoretical predictions for an electron transport model which relates both the redox levels of Q and the photosystem II optical cross section to the overall velocity of noncyclic electron flow. It is demonstrated that transfer of 15-20% of the photosystem II antenna to photosystem I may stimulate electron flow to NADP only if Q is less than 60-70% oxidized (this condition exists with our thylakoids, even at extremely low absorption fluxes, when the illumination is not specifically enriched in photosystem I absorbed wavelengths); in phosphorylated thylakoids the steady-state redox level Q is substantially shifted to a more oxidized one (measurements of this parameter using light of different wavelengths quantitatively support the idea that thylakoid phosphorylation leads to increased photosystem I and decreased photosystem II cross sections); thylakoid phosphorylation leads to stimulated noncyclic electron flow to NADP only when the increased photosystem I antenna is able to bring about large increases in the steady-state level of oxidized Q. 相似文献
20.
Data are presented which suggest that photosystem II dependent hydrogen ion accumulation and ATP synthesis can occur only after the lamellar membranes have undergone a conformational change. This membrane conformational change is detected by the electron transport dependent incorporation of diazonium benzene[35S]sulfonate into membrane components. Previously it was established that electron flux from the photosystem II primary acceptor to plastoquinone is a necessary event for the occurrence of the diazonium-detected conformational change. These studies indicate that the release of hydrogen ions during photosystem II oxidation of the primary reductant is also a necessary event for the diazonium-detected conformational change. When iodide were substituted for water (or other proton-releasing donors) as the primary reductant of system II the conformational change did not occur even though a substantial rate of electron flow from the primary acceptor to plastoquinone occurred. 相似文献