The Possible Mechanism of Photoinhibition of Purified PSI Complexes

The protecting effect of histidine on the photodamage of pigments and proteins of the isolated PSⅠ particles from the chloroplast of Spinacia oleracea L. during the strong illumination (2 300 μmol·m-2·s-1) was studied by spectroscopy and SDS-PAGE. The absorbance of PSⅠ particles decreased during the strong illumination treatment, but the decrease would be slowed down in the presence of externally added histidine after 30 min illumination. The decrease of CD （circular dichroism）signal intensities of PSⅠ particles also was slowed down by the added histidine after about 10 min illumination. The retarded protecting effect of the added histidine on the photobleaching of pigments of PSⅠ complexes implied that the mechanisms of photoinhibition of isolated PSⅠ complexes are different from early stage to later stage during the strong illumination treatment. In addition, the added histidine suppressed the decrease of 77 K fluorescence yield of PSⅠ particles during the illumination. SDS-PAGE showed that the added histidine not only protected the reaction center proteins of PSⅠ particles, but also protected other subunits of PSⅠ particles from degradation.     

Protein identification of purified particles isolated from spinach thylakoids by deoxycholate electrophoresis

Three thylakoid complexes were isolated by deoxycholate preparative electrophoresis. The protein composition of each fraction was analyzed by SDS analytical electrophoresis. No protein of the PS 1 enriched fraction (fraction 1) was found in the PS 2 enriched fraction (fraction 2) and inversely. The antenna complex (fraction 3) did not have any contamination by proteins of fraction 1 or fraction 2. Fraction 1 was mainly composed of the CP1, the reaction center complex of the PS1, and by low molecular weight proteins, previously found in other PS 1 preparations. Tentative assignments of these proteins are presented; among them are iron sulfur proteins. After analytical SDS electrophoresis of fraction 2, the reaction center complex was dissociated. Nevertheless three proteins of 50 kD, 42 kD and 35 kD were assigned to this complex. Fraction 2 contained also the three cytochromes of the thylakoid membranes: cyt f, cyt b6, cyt b559. Fraction 3 was exclusively composed of one protein pigment complex, CP2.Abbreviations SDS sodium dodecyl sulfate - PS 1 photosystem 1 - PS 2 photosystem 2 - CP1, CP2 protein pigment complexes isolated by SDS electrophoresis - cyt cytochromes - P700 primary electron donor of PS 1 - P680 primary electron donor of PS 2 - DOC deoxycholate - Q primary plastoquinone electron acceptor - CF coupling factor     

Effects of phospholipase A2 and alkaline pH on photosystem II particles isolated from spinach chloroplasts

An O₂-evolving photosystem II fraction (PS II particles) isolated from spinach ( Spinacia oleracea L.) chloroplasts by Triton X-100 was treated by phospholipase A₂ or by an alkaline pH. Phospholipase A₂ depleted the particles of all phosphatidylcholine and of a part of phosphatidylglycerol containing trans -hexadecenoic acid, and induced a parallel inactivation of the PS II activity. The protein pattern remained similar to that of the control particles. The addition of exogenous polar lipids from thylakoids could not reactivate PS II activity. Treatment of PS II particles by an alkaline pH, known to release the 33, 24 and 18 kdalton polypeptides and to inactivate PS II activity, did not affect the lipid composition. The involvement of lipids in PS II activity is discussed.     

Photochemical activities,pigment distribution and photosynthetic unit size of subchloroplast particles isolated from synchronized cells of Scenedesmus obliquus

Photosystem II (PS II) reactions of chloroplast particles show the same variations during the synchronous life cycle of Scenedesmus obliquus, strain D₃ (Gaffron Biol. Zbl. 59, 302 1939), as the whole cells they derived from. Photosystem I (PS I) reactions of whole cells and of subchloroplast particles show little or no variation in their activity, whereas PS I reactions of chloroplast particles vary like PS II reactions during the life cycle. The variation in chloroplast particles could be attributed to the change in the reoxidation capacity of plastoquinone still attached to PS I. Digitonin-treatment of chloroplast particles from Scenedesmus and subsequent sucrose density gradient separation yielded 3 distinct fractions: Fraction I contained pure PS I particles with the most efficient PS I-mediated methylviologen (MV) reduction with subsequent oxygen uptake (3 mmol O₂/mg Chl·h); no Hill reaction; and a high chlorophyll a/b ratio, and a vast amount of unbound protein xanthophyll complexes. Fraction II is enriched in PS II particles, with little PS I activity (less than 10% of the PS I particles) and a low chlorophyll a/b ratio. The activity of the water-splitting system was completely lost. This fraction must also contain most of the light-harvesting pigment system. Fraction III is also enriched in PS II with even less PS I activity, but the ratio of chlorophyll a/b is slightly higher than in whole cells and the water-splitting system is intact. -carotene was part of all fractions whereas functional xanthophylls seemed to be restricted to the PS II particles. From the constant chlorophyll P/700 ratio we had to conclude that size of the photosynthetic unit does not change during the life cycle of a synchronized Scenedesmus obliquus culture.Abbreviations DBMIB 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethyl-urea - DCPIP dichlorphenolindophenol - MV methylviologen (paraquat) - PS I photosystem I - PS II photosystem II - DPC diphenyl-carbazide     

Interaction of Photosystem 2-LHC2 Supercomplexes in Adjacent Layers of Stacked Chloroplast Thylakoid Membranes

Two Types of photosystem 2-light-harvesting complex 2 (PS2-LHC2) supercomplexes with similar pigment and protein composition were isolated directly from thylakoid membranes by sucrose density gradient centrifugation. Electron microscopy and single particle image analysis revealed the first Type as single unpaired PS2-LHC2 supercomplexes, whereas the second Type was characterized as pairs of two PS2-LHC2 supercomplexes attached together by their stromal sides. Unstacking of thylakoid membranes resulted in a spontaneous disintegration of the paired supercomplexes into single unpaired particles. A model of the organisation of the pigment-protein complexes in grana region is proposed.     

Integrity and Activity of Photosystem 2 Complexes Isolated from the Thermophilic Cyanobacterium Synechococcus Elongatus Using Various Detergents

The efficiency in selective extraction of photosystem (PS) 2 oxygen evolving complexes was compared among seven detergents. These were applied to thylakoid membranes of the thermophilic cyanobacterium Synechococcus elongatus. Used were five non-ionic detergents with one ionic and one zwitterionic for comparison. To compare the suitability and efficiency of the detergents the following properties of the extracts were examined: maximum rate of oxygen evolution with various electron acceptors, the relative variable fluorescence (F_V/F_M), the contamination of the extract with photosystem (PS) 1, and the status of the electron acceptor side of PS2 reaction centre. None of the detergents yielded a highly selective extraction of the PS2 complexes (negligible contamination with PS1) which would simultaneously display a high photochemical activity and high structural intactness. Heptylthioglucoside and dodecylmaltoside yielded the nearest approximation to the optimum result. Kinetic fluorometry was applied here for the first time to characterize the functional and structural properties of PS2 particles from cyanobacteria. This revised version was published online in August 2006 with corrections to the Cover Date.     

Isolation of an oxygen-evolving Photosystem II preparation containing only one tightly bound calcium atom from a chlorophyll b-deficient mutant of rice

Oxygen-evolving Photosystem II (PS II) particles were prepared from the thylakoid membranes of a chlorophyll b-less rice mutant, which totally lacks light-harvesting chlorophyll a/b proteins, after solubilization with β-octylglucoside. The preparation was essentially free of Photosystem I as judged from its low-temperature fluorescence spectrum and polypeptide composition. The PS II particles contained all the major subunit polypeptides of the PS II reaction center core complexes and the three extrinsic proteins related to oxygen evolution. The relative abundances of the 33, 21 and 15 kDa proteins were 100, 64 and 20%, respectively, of the corresponding proteins in the mutant thylakoids. The chlorophyll-to-Q_A ratio was 53 and there was only one bound Ca²⁺ per Q_A. Thus, one of the two bound Ca²⁺ present in the oxygen-evolving PS II membrane preparations from wild-type rice (Shen J.-R., Satoh, K. and Katoh, S. (1988) Biochim. Biophys. Acta 933, 358–364) is missing. The mutant PS II particles were highly active in oxygen evolution in the absence of exogenously added Ca²⁺, although addition of 5 mM Ca²⁺ enhanced the activity by 30%. When the 21 and 15 kDa proteins were supplemented to the particles, the Ca²⁺-effect disappeared and the rate of oxygen evolution increased to a level exceeding 1000 μmol O₂ per mg chlorophyll per h. The results indicate that the number of Ca²⁺ needed to promote a high rate of oxygen evolution is one per PS II in higher plants.     

Photoinactivation of photosystem II complexes and photoprotection by non-functional neighbours in Capsicum annuum L. leaves

Leaf segments from Capsicum annuum plants grown at 100 micromol photons m(-2) s(-1) (low light) or 500 micromol photons m(-2) s(-1) (high light) were illuminated at three irradiances and three temperatures for several hours. At various times, the remaining fraction (f) of functional photosystem II (PS II) complexes was measured by a chlorophyll fluorescence parameter (1/Fo -1/Fm, where Fo and Fm are the fluorescence yields corresponding to open and closed PS II traps, respectively), which was in turn calibrated by the oxygen yield per saturating single-turnover flash. During illumination of leaf segments in the presence of lincomycin, an inhibitor of chloroplast-encoded protein synthesis, the decline of f from 1.0 to about 0.3 was mono-exponential. Thereafter, f declined much more slowly, the remaining fraction (approximately equals 0.2) being able to survive prolonged illumination. The results can be interpreted as being in support of the hypothesis that photoinactivated PS II complexes photoprotect functional neighbours (G. Oquist et al. 1992, Planta 186: 450-460), provided it is assumed that a photoinactivated PS II is initially only a weak quencher of excitation energy, but becomes a much stronger quencher during prolonged illumination when a substantial fraction of PS II complexes has also been photoinactivated. In the absence of lincomycin, photoinactivation and repair of PS II occur in parallel, allowing f to reach a steady-state value that is determined by the treatment irradiance, temperature and growth irradiance. The results obtained in the presence and absence of lincomycin are analysed according to a simple kinetic model which formally incorporates a conversion from weak to strong quenchers, yielding the rate coefficients of photoinactivation and of repair for various conditions, as well as gaining an insight into the influence off on the rate coefficient of photoinactivation. They demonstrate that the method is a convenient alternative to the use of radiolabelled amino acids for quantifying photoinactivation and repair of PS II in leaves.     

A single step separation of PS 1, PS 2 and chlorophyll-antenna particles from spinach chloroplasts

After solubilization of photosynthetic membranes by digitonin, three main protein pigment complexes were isolated by electrophoresis with deoxycholate as detergent.The band with the slowest mobility, fraction 1, had PS 1 activity and was devoid of PS 2 activity. This fraction was four times enriched in P700 when compared with chloroplasts. Fraction 1 had little chl b, a long wavelength absorption maximum in the red, a maximum of low temperature emission fluorescence at 730nm, and a circular dichroism spectrum characteristic of PS 1 enriched fraction.Fraction 2 exhibited a PS 2 activity and no PS 1 activity. It was enriched five times in PS 2 reaction centre and had little chl b and carotenoids. The absorption maximum was at 674 nm and the low temperature fluorescence emission maximum was at 700 nm. Fraction 2 might be useful PS 2 enriched particle because of the great stability of this fraction with regard to photochemical activity and also rapidity and simplicity of its preparation.Fraction 3, which had the fastest migration, was devoid of photochemical activities; It was rich in chl b and had the fluorescence and the circular dichroism spectrum characteristic of an antenna complex.Abbreviations PS 1 (2) photosystem 1 (2) - chl chlorophyll - car carotenoid - Q primary plastoquinone electron acceptor - P700 primary electron donor of PS 1 - P680 primary electron donor of PS 2 - K₃Fe(CN)₆ potassium ferricyanide - DCMU dichlorophenyldimethylurea - DCPIP dichlorophenolindophenol - DPC diphenyl-carbazide     

Correlation of photosystem-II complexes with exoplasmatic freeze-fracture particles of thylakoids of the cyanobacterium Synechococcus sp.

The supramolecular structure of the exoplasmic freeze-fracture particles of thylakoids of the thermophilic cyanobacterium Synechococcus sp. is compared with that of isolated photosystem-II complexes. The in-situ EF particles are scattered on the thylakoids or organized in rows of variable length; the latter aligned particles measure 10 nmx20 nm and are separated perpendicular to their long axis into two parts. We propose that they represent dimers composed of two monomeric 10-nm EF particles side by side. Isolated photosystem (PS)II particles correspond in size to the monomeric 10-nm EF particles as analysed by negative contrast and freeze-fracture electron microscopy. Dimeric PSII particles, very similar to the in-situ 10 nmx20 nm EF particles, are obtained after incorporation of purified PSII complexes into liposomes made from phospholipid and cholesterol. Each monomeric complex consists of the reaction center, the water-splitting system, the chlorophyll antennae and phycobilisome-binding polypeptides. We propose that the dimeric complexes bind one hemidiscoidal phycobilisome at their domains exposed to the external side of the thylakoids. The implications of this arrangement of the PSII-phycobilisome complexes within the thylakoids upon excitation-energy distribution are discussed.Abbreviations EF exoplasmic fracture face - LDS lithium dodecyl sulfate - PAGE polyacrylamide gel electrophoresis - PS photosystem - SDS sodium dodecyl sulfate - SPC-buffer 0.5 M sucrose, 0.5 M K₂HPO₄/KH₂PO₄, 0.3 M Nacitrate, pH 7.0 This study is dedicated to Professor W. Nultsch on the occasion of his 60th birthday.     

Photosystem I from the unusual cyanobacterium Gloeobacter violaceus

Photosystem I (PS I) from the primitive cyanobacterium Gloeobacter violaceus has been purified and characterised. Despite the fact that the isolated complexes have the same subunit composition as complexes from other cyanobacteria, the amplitude of flash-induced absorption difference spectra indicates a much bigger antenna size with about 150 chlorophylls per P700 as opposed to the usual 90. Image analysis of the PS I preparation from Gloeobacter reveals that the PS I particles exist both in a trimeric and in a monomeric form and that their size and shape closely resembles other cyanobacterial PS I particles. However, the complexes exhibit a higher molecular weight as could be shown by gel filtration. The preparation contains novel polypeptides not related to known Photosystem I subunits. The N-terminal sequence of one of those polypeptides has been determined and reveals no homology to known or hypothetical proteins. Immunoblotting shows a cross-reaction of three of the polypeptide bands with an antibody raised against the major LHC from the diatom Cyclotella cryptica. Electron microscopy reveals a novel T-shaped complex which has never been observed in any other cyanobacterial PS I preparation. 77 K spectra of purified PS I show an extreme blue-shift of the fluorescence emission, indicating an unusual organisation of the PS I antenna system in Gloeobacter. This revised version was published online in June 2006 with corrections to the Cover Date.     

Reversible particle movements associated with unstacking and restacking of chloroplast membranes in vitro

Freeze-fracture and freeze-etch techniques have been employed to study the supramolecular structure of isolated spinach chloroplast membranes and to monitor structural changes associated with in vitro unstacking and restacking of these membranes. High-resolution particle size histograms prepared from the four fracture faces of normal chloroplast membranes reveal the presence of four distinct categories of intramembranous particles that are nonrandomly distributed between grana and stroma membranes. The large surface particles show a one to one relationship with the EF-face particles. Since the distribution of these particles between grana and stroma membranes coincides with the distribution of photosystem II (PS II) activity, it is argued that they could be structural equivalents of PS II complexes. An interpretative model depicting the structural relationship between all categories of particles is presented. Experimental unstacking of chloroplast membranes in low-salt medium for at least 45 min leads to a reorganization of the lamellae and to a concomitant intermixing of the different categories of membrane particles by means of translational movements in the plane of the membrane. In vitro restacking of such experimentally unstacked chloroplast membranes can be achieved by adding 2-20 mM MgCl2 or 100-200 mM NaCl to the membrane suspension. Membranes allowed to restack for at least 1 h at room temperature demonstrate a resegregation of the EF-face particles into the newly formed stacked membrane regions to yield a pattern and a size distribution nearly indistinguishable from the normally stacked controls. Restacking occurs in two steps: a rapid adhesion of adjoining stromal membrane surfaces with little particle movement, and a slower diffusion of additional large intramembranous particles into the stacked regions where they become trapped. Chlorophyll a:chlorophyll b ratios of membrane fraction obtained from normal, unstacked, and restacked membranes show that the particle movements are paralleled by movements of pigment molecules. The directed and reversible movements of membrane particles in isolated chloroplasts are compared with those reported for particles of plasma membranes.     

Influence of clinorotation on some parameters of photosynthetic apparatus

This study aimed to examine the electron transport rates in the thylakoids, isolated from leaves of pea plants grown under clinorotation and in vertical control, to measure the chlorophyll a/b (Chl a/b) ratio in such thylakoids and in photosystem I (PSI) particles isolated from them, to elucidate if there are any differences in changes of PS II activity in thylakoids and Chl a/b ratio in PS I particles under phosphorylation of polypeptides of thylakoid pigment-protein complexes.     

Isolation and characterization of Photosystem I from two strains of the marine oxychlorobacterium Prochlorococcus

Photosystem I (PS I) complexes from two strains of the marine photosynthetic prokaryote Prochlorococcus, MED4 (= clone CCMP1378) and SS120 (= clone CCMP1375), were isolated by centrifugation on sucrose gradients after detergent treatment. The PS I-enriched fractions of both strains contained about 100 chlorophyll molecules per P700. Electron microscopy showed that the PS I complexes were in a trimeric form. The characteristic long wavelength fluorescence emission of PS I at 77 K, currently observed in chloroplasts and most cyanobacteria was absent both in intact cells and in PS I preparations of both strains. The major proteins of the PS I-enriched fractions were identified immunologically as PsaA and PsaB. Two proteins with apparent molecular masses of about 21 and 25 kDa were present in PS I preparations of Prochlorococcus, whereas the small PS I subunits in cyanobacteria all have molecular masses below 18 kDa. The 25 kDa protein showed a strong cross-reaction with a heterologous antibody against PsaL. Relatedness of the 21 kDa protein to PsaF was demonstrated by internal protein sequencing. Although only trace amounts of the major divinyl-Chl a/b-binding antenna complexes were present in the PS I preparations, significant amounts of divinyl-Chl b were observed in this fraction. The putative organization of this Chl b in PS I is discussed.     

The Regulation and Distribution of LHC-I and LHCP2 between the Photosystem I and Photosystem II

Evidences were provided in this paper that the relative distribution of chl-protein complexes of PSⅠ and PSⅡ could be regulated by Mg2+. addition of Mg2+ led to decrease in the amount of chl-protein complexes of PSⅠ and increase in the amount of chl-protein in complexes of PSⅡ. There was no effect of Mg2+ on the spectral property of LHCP1, but the addition of Mg2+ could change the spectral property of LHCP2 so that it became similar to that of the LHC-Ⅰ. CPIa2 was a complex of reaction centre of PSⅠ and LHC-I. LHC-I might be contacted specially with LHCP2 in chloroplast membranes. Addition of Mg2+ probably cansed the motion of LHC-I from PSⅠ to PSⅡ and became more closely connected with LHCP2. The relative amount of CPIa2, CPIa1, LHCP1 and LHCP2 in chloroplast membranes could be regulated by different light intensity. There were more CPIa2, LHCP1 and less LHCP2 in chloroplast membranes from the shade plant Malaxis monophyllos and sunflower grown under weak light, both of them lacked equally CPIa1. There were less CPIa2, LHCP1 and more LHCP2 in the sun plant spinach and sunflower grown under strong light, and they possessed equally CPIa1 chl-protein complexes. It is suggested that LHCP1 and LHCP2 are different light-harvesting Chl-protein complexes. The LHC-I and LHCP2 are mobile light-harvesting chl-protein complexes and shuttle back and forth between PSⅠ and PSⅡ They play an important role in the regulation and distribution of excitation energy between the two photosystems.     

Presenilin 2 interacts with sorcin, a modulator of the ryanodine receptor

Perturbed Ca(2+) homeostasis is a common molecular consequence of familial Alzheimer's disease-linked presenilin mutations. We report here the molecular interaction of the large hydrophilic loop region of presenilin 2 (PS2) with sorcin, a penta-EF-hand Ca(2+)-binding protein that serves as a modulator of the ryanodine receptor intracellular Ca(2+) channel. The association of endogenous sorcin and PS2 was demonstrated in cultured cells and human brain tissues. Membrane-associated sorcin and a subset of the functional PS2 complexes were co-localized to a novel subcellular fraction that is distinctively positive for calcineurin B. Sorcin was found to interact with PS2 endoproteolytic fragments but not full-length PS2, and the sorcin/PS2 interaction was greatly enhanced by treatment with the Ca(2+) ionophore A23187. Our findings reveal a molecular link between PS2 and intracellular Ca(2+) channels (i.e. ryanodine receptor) and substantiate normal and/or pathological roles of PS2 in intracellular Ca(2+) homeostasis.     

Delayed fluorescence observed in the nanosecond time region at 77 K originates directly from the photosystem II reaction center

The excited-state dynamics of delayed fluorescence in photosystem (PS) II at 77 K were studied by time-resolved fluorescence spectroscopy and decay analysis on three samples with different antenna sizes: PS II particles and the PS II reaction center from spinach, and the PS II core complexes from Synechocystis sp. PCC 6803. Delayed fluorescence in the nanosecond time region originated from the 683-nm component in all three samples, even though a slight variation in lifetimes was detected from 15 to 25 ns. The relative amplitude of the delayed fluorescence was higher when the antenna size was smaller. Energy transfer from the 683-nm pigment responsible for delayed fluorescence to antenna pigment(s) at a lower energy level was not observed in any of the samples examined. This indicated that the excited state generated by charge recombination was not shared with antenna pigments under the low-temperature condition, and that delayed fluorescence originates directly from the PS II reaction center, either from chlorophyll a(D1) or P680. Supplemental data on delayed fluorescence from spinach PS I complexes are included.     

The Drosophila position-specific antigens are a family of cell surface glycoprotein complexes

Position-specific (PS)1 and PS2 monoclonal antibodies bind non-uniformly to the mature wing imaginal disc of Drosophila with respect to the boundary separating the dorsal and ventral developmental compartments. PS1 antibodies preferentially recognize dorsal cells, PS2 antibodies ventral cells. Antibodies of the two classes extract distinct sets of glycoproteins from an imaginal disc lysate. PS3 antibodies bind to both dorsal and ventral disc cells and extract both PS1 and PS2 glycoprotein sets together with an additional component. We show that the PS antigens are related multimeric glycoprotein complexes on the cell surface. PS3 antibodies recognize a glycoprotein present in all complexes, while PS1 and PS2 antibodies recognize unique components of their own complexes. Spatial and temporal correlations suggest the molecules may have a function in development.     

Delayed fluorescence observed in the nanosecond time region at 77 K originates directly from the photosystem II reaction center

The excited-state dynamics of delayed fluorescence in photosystem (PS) II at 77 K were studied by time-resolved fluorescence spectroscopy and decay analysis on three samples with different antenna sizes: PS II particles and the PS II reaction center from spinach, and the PS II core complexes from Synechocystis sp. PCC 6803. Delayed fluorescence in the nanosecond time region originated from the 683-nm component in all three samples, even though a slight variation in lifetimes was detected from 15 to 25 ns. The relative amplitude of the delayed fluorescence was higher when the antenna size was smaller. Energy transfer from the 683-nm pigment responsible for delayed fluorescence to antenna pigment(s) at a lower energy level was not observed in any of the samples examined. This indicated that the excited state generated by charge recombination was not shared with antenna pigments under the low-temperature condition, and that delayed fluorescence originates directly from the PS II reaction center, either from chlorophyll a_D1 or P680. Supplemental data on delayed fluorescence from spinach PS I complexes are included.     

Assembly of photosystem I. II. Rubredoxin is required for the in vivo assembly of F(X) in Synechococcus sp. PCC 7002 as shown by optical and EPR spectroscopy

The rubA gene was insertionally inactivated in Synechococcus sp. PCC 7002, and the properties of photosystem I complexes were characterized spectroscopically. X-band EPR spectroscopy at low temperature shows that the three terminal iron-sulfur clusters, F(X), F(A), and F(B), are missing in whole cells, thylakoids, and photosystem (PS) I complexes of the rubA mutant. The flash-induced decay kinetics of both P700(+) in the visible and A(1)- in the near-UV show that charge recombination occurs between P700(+) and A(1)- in both thylakoids and PS I complexes. The spin-polarized EPR signal at room temperature from PS I complexes also indicates that forward electron transfer does not occur beyond A(1). In agreement, the spin-polarized X-band EPR spectrum of P700(+) A(1)- at low temperature shows that an electron cycle between A(1)- and P700(+) occurs in a much larger fraction of PS I complexes than in the wild-type, wherein a relatively large fraction of the electrons promoted are irreversibly transferred to [F(A)/F(B)]. The electron spin polarization pattern shows that the orientation of phylloquinone in the PS I complexes is identical to that of the wild type, and out-of-phase, spin-echo modulation spectroscopy shows the same P700(+) to A(1)- center-to-center distance in photosystem I complexes of wild type and the rubA mutant. In contrast to the loss of F(X), F(B), and F(A), the Rieske iron-sulfur protein and the non-heme iron in photosystem II are intact. It is proposed that rubredoxin is specifically required for the assembly of the F(X) iron-sulfur cluster but that F(X) is not required for the biosynthesis of trimeric P700-A(1) cores. Since the PsaC protein requires the presence of F(X) for binding, the absence of F(A) and F(B) may be an indirect result of the absence of F(X).