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1.
The photoreactions mediated by Photosystem II at low temperatures were examined in subchloroplast particles enriched in Photosystem II to determine if the Photosystem II activity was independent of C-550 in such particle preparations. Two types of Photosystem II particle preparations were tested, one enriched in chlorophyll b and the other purified further to eliminate the chlorophyll b. The Photosystem II-mediated photoreactions at low temperature were the same in both of the Photosystem II particle preparations as they were in normal chloroplasts. C-550 acted as if it were the primary electron acceptor of Photosystem II in all of the experiments performed. 相似文献
2.
3.
Wakamatsu Kunimitsu; Ikehara Norikatsu; Nishimura Mitsuo 《Plant & cell physiology》1974,15(4):601-610
The light-induced absorbance change at 515 nm, light-inducedhydrogen ion uptake and ATP formation were compared in chloroplastsand different types of sonicated subchloroplast particles. Noparallel relationship among the activities for ATP formation,hydrogen ion uptake and the 515-nm change was observed in differenttypes of preparations. NH4Cl inhibited ATP formation in chloroplastsbut had little effect on subchloroplast particles. In contrast,the light-induced hydrogen ion uptake was inhibited by NH4Clin a similar manner. Tetraphenylboron (TPB), at 1 µM, inhibited ATP formationby about 30% in both chloroplasts and subchloroplast particles.In the presence of TPB, ATP formation in chloroplasts was stronglyinhibited by NHC4Cl, but in subchloroplast particles the additionalinhibitory effect of NH4Cl was small. A synergistic inhibitionof photophosphorylation by valinomycin plus NH4Cl was much clearer.Although acceleration of the recovery of the 515-nm change byNH4Cl or valinomycin was moderate, the 515-nm change virtuallydisappeared when NH4Cl and valinomycin were added simultaneously. Although the membrane potential has a major role as the principaldriving force for ATP formation in subchloroplast particles,the simultaneous abolishment of the pH gradient and membranepotential may be required to uncouple ATP formation.
1Present address: Fukuoka Women's University, Kasumigaoka, Fukuoka813, Japan.
2Present address: Ryukyu University, Naha, Okinawa 903, Japan. (Received February 5, 1974; ) 相似文献
4.
The inhibition of the oxidase and respiratory nitrate reductase activity in membrane preparations from Klebsiella aerogenes by 2-n-heptyl-4-hydroxyquinoline-N-oxide (HQNO) has been investigated. Addition of HQNO only slightly affected the aerobic steady-state reduction of cytochrome b559 with NADH, but caused a significantly lower nitrate reducing steady-state of this cytochrome. The changes in the redox states of the cytochromes during a slow transition from anaerobic to aerobic conditions in the presence and absence of HQNO showed that the inhibition site of HQNO is located before cytochrome d. Inhibition patterns obtained upon titration of the NADH oxidase and NADH nitrate reductase activity with HQNO indicated one site of inhibitor interaction in the NADH nitrate reductase pathway and suggested a multilocated inhibition of the NADH oxidase pathway. Difference spectra with ascorbate-dichlorophenolindophenol as electron donor indicated the presence of a cytochrome b563 component which was not oxidized by nitrate, but was rapidly oxidized by oxygen. The latter oxidation was prevented by HQNO. A scheme for the electron transport to oxygen and nitrate is presented. In the pathway to oxygen, HQNO inhibits both at the electron-accepting side of cytochrome b559 and at the electron-donating side of cytochrome b563, whereas in the pathway to nitrate, inhibition occurs only at the electron-accepting side of cytochrome b559. 相似文献
5.
Photosystem II particles have been prepared from spinach and Chlamydomonas reinhardii CW 15 thylakoids. Photosynthetic electron transport in these particles is inhibited by phenolic compounds like dinoseb, but not by atrazine and diuron. The labeling patterns obtained by photoaffinity labels derived from either atrazine (azido-atrazine) or the phenolic herbicide dinoseb (azido-dinoseb) were compared in photosystem II particles and thylakoids. Whereas azido-atrazine in thylakoids of spinach as well as of Chlamydomonas labels a 32-kilodalton peptide, this label does not react in photosystem II particle preparations. Azido-dinoseb, however, labels both the thylakoid membranes and the particles, predominantly polypeptides in the 40-53 kilodalton molecular weight region. Since the latter polypeptides are probably part of the reaction center of photosystem II, it is suggested that phenolic compounds have their inhibition site within the reaction center complex. This indicates that the atrazine-binding 32-kilodalton peptide is either absent or functionally inactive in photosystem II particles, whereas the phenol inhibitor-binding peptides are not. 相似文献
6.
D. J. Richardson D. J. Kelly J. B. Jackson S. J. Ferguson K. Alef 《Archives of microbiology》1986,146(2):159-165
The effects of various electron transport inhibitors upon the rates of reduction NO
3
-
, dimethyl sulphoxide (DMSO) and N2O in anaerobic suspensions of Rhodobacter capsulatus have been studied. A new method for the determination of the rates of reduction of these auxiliary oxidants in intact cells is presented, based on the proportionality observed between the concentration of oxidant and the duration of the electrochromic carotenoid bandshift. For NO
3
-
and N2O good agreement was found between rates of reduction determined using electrodes and those determined by the electrochromic method.Myxothiazol and antimycin A had no effect on the rates of reduction of NO
3
-
and DMSO suggesting that the cytochrome b/c
1complex is not involved in electron transport to these oxidants. 2-n-heptyl-4-hydroxyquinoline-N-oxide (HOQNO) inhibited at two sites, one within the cytochrome b/c
1complex and the other on the nitrate reducing pathay, but had no effect on electron transport to N2O or DMSO. In both intact cells and cell free extracts, HOQNO had no effect on the nitrate dependent re-oxidation of reduced methylviologen (MVH2), a direct electron donor to nitrate reductase.Our data are consistent with a branch point for the auxiliary electron transport pathways at the level of the ubiquinone pool.Non-standard abbreviations HOQNO
2-n-heptyl-4-hydroxyquinoline-N-oxide
- TMAO
trimethylamine-N-oxide
- DMSO
dimethyl-sulphoxide
-
membrane potential
- MVH2
reduced methyl viologen 相似文献
7.
The effect of reversible extraction of Mn on ESR signal II arising from the oxidized secondary electron donor (Z+) and the ESR doublet signal related to reduced spin-coupled pheophytin (pheo -) and "primary" electron acceptor (PA- -Fe (2+)) has been studied in oxygen evolving preparations of the photosystem 2. It is demonstrated that Mn extraction does not affect both dark and photoinduced ESR signal II and ESR doublet. A conclusion is made that Mn is not a component of the secondary electron donor Z of the Photosystem 2 and its complete removal has no effect on the exchange interaction of Pheo(-) and the PQ(-) -Fe(2+) complex. 相似文献
8.
We have studied the kinetics of binding of the menaquinol analog 2-n-heptyl-4-hydroxyquinoline-N-oxide (HOQNO) by fumarate reductase (FrdABCD) using the stopped-flow method. The results show that the fluorescence of HOQNO is quenched when HOQNO binds to FrdABCD. The observed quenching of HOQNO fluorescence has two phases and it can be best fitted to a double exponential equation. A two-step equilibrium model is applied to describe the binding process in which HOQNO associates with FrdABCD by a fast bimolecular step to form a loosely bound complex; this is subsequently converted into a tightly bound complex by a slow unimolecular step. The rates of the forward and the reverse reactions for the first equilibrium (k1 and k2) are determined to be k1 = (1.1 +/- 0.1) x 10(7) M-1.s-1, and k2 = 6.0 +/- 0.6 s-1, respectively. The dissociation constants of the first equilibrium (Kd1 = k2/k1) is calculated to be about 550 nM. The overall dissociation constant for the two-step equilibrium, Kd overall = Kd1/[1+ (1/Kd2)], is estimated to be < or = 7 nM. Comparison of the kinetic parameters of HOQNO binding by FrdABCD and by dimethyl sulfoxide reductase provides important information on menaquinol binding by these two enzymes. 相似文献
9.
Anisotropic properties of pea chloroplasts, subchloroplast fragments (photosystem 1 particles and pigment-protein complexes) and the blue-green algae oriented in polyacrylamide gel were investigated. It was shown that linear dichroism spectra of chloroplasts are the superposition of the corresponding spectra for the main light harvesting complex (HMLC) and P 700 chlorophyll a--protein complex (CP 1). Anisotropic properties of the photosystem 1 particles and blue-green algae are mainly caused by CP 1 anisotropy. Qy-transition moments tend to perpendicular orientation to the membrane plane for the Chl. b 649, Chl. a 660 and parallel orientation--for Chl. b 654, Chl. a 682. The degree of Qy-transition moments parallel orientation is higher for the longwave forms (Chl. a 690, Chl. a 702, Chl. a 712), than for the shortwave ones and coincides with this degree for the reaction centre pigment P 700 transition moment. It is suggested that the specific orientation of the pigment-protein complexes in the chloroplast membrane is important for the regulation of the spillover between two photosystems. 相似文献
10.
Phycobiliproteins obtained by dissociation of phycobilisomes were reassociated in vitro with intact thylakoids or isolated photosystems I and II preparations obtained from cyanophytes (prokaryotes) or green algae (eukaryotes) to form bound phycobilisome complexes. Energy transfer from Fremyella diplosiphon phycobiliproteins to chlorophyll a of reaction centers I and II was measured in: complexes containing intact thylakoids of the cyanophytes F. diplosiphon or Anacystis nidulans and the eukaryotic algae Euglena gracilis and mutants of Chlamydomonas reinhardtii; complexes containing isolated photosystem II particles of A. nidulans or C. reinhardtii; and complexes containing reaction center I of F. diplosiphon or C. reinhardtii. Energy transfer from phycoerythrin to chlorophyll a of photosystem II could be demonstrated in complexes containing phycobilisomes bound to cyanophyte thylakoids or isolated photosystem II particles of A. nidulans or C. reinhardtii. Bound phycobilisomes did not transfer energy to photosystem II within green algae thylakoids containing altered forms of light-harvesting chlorophyll a/b-protein complex (LHC) II antenna, reduced amounts of LHC II, or chlorophyll b, or chlorophyll b-less mutants, nor to chlorophyll a of photosystem I of intact thylakoids or isolated reaction centers. We conclude that phycobilisomes can form a specific and functional association with photosystem II particles of both cyanophytes and eukaryotic thylakoids. This interaction appears to be hindered by the presence of LHC II antenna in the eukaryotic thylakoids. 相似文献
11.
Regulation of excitation energy distribution in subchloroplast particles: photosystem I 总被引:3,自引:0,他引:3
Mono- and divalent cations were found to increase the transfer of excitation energy within Photosystem I from the light-harvesting chlorophyll a molecules to P700. The P700-chlorophyll a protein of Shiozawa et al. (J. A. Shiozawa, R. S. Alberte, and J. P. Thornber, 1974, Arch. Biochem. Biophys.165, 388–397) was used for these studies. Cations stimulated the quantum yields for electron transport when the light-harvesting chlorophyll a molecules were irradiated. They also decreased chlorophyll a fluorescence. Half-maximal effects were observed at 0.5–0.6 mm for divalent cations and at 5–6 mm for monovalent cations. Triton X-100, 0.02%, also increased energy transfer. The increases in energy transfer are due to an intramolecular conformational change in the protein. A structural change is involved, since there is a correlation between the cation-induced changes in energy transfer and increases in 90 ° light scattering. However, there was no change in the molecular weight upon the addition of MgCl2. The molecular weight, as determined by gel filtration, was 105,000 in the presence of 0.05% Triton X-100. On the other hand, circular dichroism measurements showed an increase in the α-helical content from 51 to 63% when 5 mm MgCl2 was added. Changes in the absorption spectra were also observed. We believe that the cation regulation of Photosystem I activity provides a fine-tuning mechanism for the regulation of energy transfer. 相似文献
12.
Photoreduction of 2,6-dichlorophenolindophenol by diphenylcarbazide: a photosystem 2 reaction catalyzed by tris-washed chloroplasts and subchloroplast fragments 总被引:4,自引:13,他引:4
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The use of diphenylcarbazide as an electron donor coupled to the photoreduction of 2,6-dichlorophenolindophenol by tris-washed chloroplasts or subchloroplast fragments provides a simple and sensitive assay for photosystem 2 of chloroplasts. By varying the concentration of tris buffer at pH 8.0 during an incubation period it is shown that the destruction of oxygen evolution activity is accompanied by a corresponding emergence of an ability to photooxidize diphenylcarbazide, as evidenced by absorbance changes due to diphenylcarbazide at 300 nm. The temperature-sensitive oxidation of diphenylcarbazide is inhibited by DCMU and by high ionic strengths. This activity appears to measure the primary photochemical reaction of photosystem 2. 相似文献
13.
After a complete removal of Mn from pea subchloroplast photosystem-II (PS II) preparations the electron phototransfer and oxygen evolution are restored upon addition of Mn2+ and Ca2+. Pre-illumination of the sample in the absence of Mn2+ leads to photoinhibition (PI) — irreversible loss of the capability of PS II to be reactivated by Mn2+. The effect of PI is considerably decreased in the presence of Mn2+ (4 Mn atoms per reaction center of PS II) and it is increased in the presence of ferricyanide or p-benzoquinone revealing the oxidative nature of the photoeffect. PI results in suppression of oxygen evolution, variable fluorescence, photoreduction of 2,6-dichlorophenol indophenol from either water or diphenylcarbazide. However, photooxidation of chlorophyll P680, the primary electron donor of PS II as well as dark and photoinduced EPR signal II (ascribed to secondary electron donors D
1 and Z) are preserved. PI is accompanied by photooxidation of 2–3 carotenoid molecules per PS II reaction center (RC) that is accelerated in the presence of ferricyanide and is inhibited upon addition of Mn2+ or diuron. The conclusion is made that PI in the absence of Mn leads to irreversible oxidative inactivation of electron transfer from water to RC of PS II which remains photochemically active. A loss of functional interaction of RC with the electron transport chain as a common feature for different types of PS II photoinhibition is discussed.Abbreviations A
photoinduced absorbance changes
- DPC
diphenylcarbazide
- DPIP
2,6-dichlorophenol indophenol
-
F
o
constant fluorescence of chlorophyll
- F
photoinduced changes of Chl fluorescence yield
- Mn
manganese
- P680
the primary electron donor in PS II
- PI
photoinhibition
- PS II
photosystem II
-
Q
the primary (quinone) electron acceptor in PS II
- RC
reaction center 相似文献
14.
15.
A model for the photochemical apparatus of photosynthesis is presented which accounts for the fluorescence properties of Photosystem II and Photosystem I as well as energy transfer between the two photosystems. The model was tested by measuring at - 196 degrees C fluorescence induction curves at 690 and 730 nm in the absence and presence of 5mMMgCl2 which presumably changes the distrubution of excitation energy between the two photosystems. The equations describing the fluorescence properties involve terms for the distribution of absorbed quanta, alpha, being the fraction distributed to Photosystem I, and beta, the fraction to Photosystem II to Photosystem I, KT(II yields I). The data, analyzed within the context of the model, permit a direct comparison of alpha and kt(II yields I) in the absence (minus) and presence (+) of Mg-2+ :alpha minus/alpha-+ equals 1.2 and k-minus t)II yields I)/K-+T(II yields I) equal to 1.9. If the criterion that alpha + beta equal to 1 is applied absolute values can be calculated: in the presence of Mg-2+, alpha-+ equal to 0.27 and the yield of energy transfer, phi-+ t(II yields I) varied the presence of Mg-2+, alpha-+ equal to 0.27 and the yield of energy transfer, phi-+ t(II yields I) varied from 0.065 when the Photosystem II reaction centers were all open to 0.23 when they were closed. In the absence of Mg-2+, alpha-minus equal to 0.32 and phi t(II yields I) varied from 0.12 to 0.28. The data were also analyzed assuming that two types of energy transfer could be distinguished; a transfer from the light-harvesting chlorophyll of Photosystem II to Photosystem I, kt(II yields I), and a transfer from the reaction centers of Photosystem II to Photosystem I, kt(II yields I). In that case alpha-minus/alpha+ equal to 1.3, k-minus t(II yields I)/k+ t(II yields I)equal to 1.3 and k-minus t(II yields I) equal to 3.0. It was concluded, however, that both of these types of energy transfer are different manifestations of a single energy transfer process. 相似文献
16.
17.
The effects of UV radiation on the low temperature fluorescenceand primary photochemistry of PSII and PSI of spinach chloroplastswere studied. Fluorescence induction curves at 196°Cwere measured at 695 nm for PSII fluorescence and at 730 nmfor PSI fluorescence to determine both the initial Fo and finalFM levels. The primary photochemistry of PSII was measured asthe rate of photoreduction of C-550 at 196°C, thatof PSI as the rate of photooxidation of P700 at 196°C.The results were analyzed in terms of a model for the photosyntheticapparatus which accounts for the yields of fluorescence andprimary photochemistry. According to this analysis UV radiationincreases nonradiative decay processes at the reaction centerchlorophyll of PSII. However, the effect of UV radiation isnot uniform throughout the sample during irradiation so thataccount must be taken of the fraction of PSII reaction centerswhich have been irradiated at any given time. UV radiation alsoinactivates P700 and causes a slight increase in nonradiativedecay in the antenna chlorophyll of PSI. All fluorescence ofvariable yield, FV = FMFo, at 730 nm is due to energytransfer from PSII to PSI so that the sensitivity of Fv to UVradiation is the same at 730 and 695 nm.
1Present address: Department of Biology, Faculty of Science,Toho University, Narashino, Chiba 275, Japan.
2Present address: Central Research Laboratories, Fuji PhotoFilm Co., Ltd., 105 Mizonuma, Asaka-Shi, Saitama 351, Japan. (Received September 10, 1975; ) 相似文献
18.
The modes of actions of six inhibitors on the electron transportsystem in the vicinity of system II in chloroplasts were studied. The first group, including piericidin A, ioxynil and broxynil,showed relatively simple modes of action on the Hill reaction,fluorescence of chlorophyll and the photobleaching of photosyntheticpigments, which are similar to the action of DCMU. As compared with inhibitors of the first group, the inhibitoryactions of salicylaldoxime, antimycin A and azide on the Hillreaction were more complicated in that they were influencedmore strongly by reaction conditions, i.e. duration of incubation,pH of the reaction mixture and illumination of chloroplasts.Inhibitors of the second group suppressed the rise in fluorescencein the induction period. However, this effect was not observedin the presence of DCMU or dithionite. Salicylaldoxime and azidewere effective in inducing photobleaching of photosyntheticpigments, whereas antimycin A inhibited the photobleaching inducedby ferricyanide or CCCP. Inhibition sites of the inhibitors in the first group are assumedto be similar to that of DCMU, whereas the inhibitors in thesecond group are effective in blocking electron transport onthe oxidizing side of system II between the primary electrondonor of system II and an intermediary electron carrier whichreceives electrons from artificial electron donors for systemII. (Received October 30, 1971; ) 相似文献
19.
P Riccio E Marra S Passarella E Quagliariello 《Bollettino della Società italiana di biologia sperimentale》1979,55(23):2412-2418
The binding of 3H-NQNO in submitochondrial particles was determined by measuring the radioactivity in the supernatants as well as in the sediments after centrifugation of particles suspensions containing different amounts of 3H-NQNO. From the binding data Scatchard plots were constructed showing a large amount of aspecific binding depending on the particles preparation and concentration. In the presence of saturating concentrations of either antimycin or unlabelled NQNO (2-n-Nonyl-4-hydroxy-quinolinee-N-oxide) that remove or prevent the specific binding of 3H-NQNO, it is possible to evaluate the aspecific component of 3H-NQNO binding and to subtracte it from the experimental binding data by graphyc correction according to (3). The straight line from the corrected points gives the specific binding parameters: number of specific binding sites: about 0,5 moles 3H-NQNO/ moles cytochrome b and KD= 50 nM. 相似文献