Characterization of a D1‐D2‐cyt b‐559 complex containing 4 chlorophyll a/2 pheophytin a isolated with the use of MgSO4

Yruela, Inmaculada; Kan, P. Van; Müller, Martín; Holzwarth, Alfred R.

doi:10.1016/0014-5793(94)80377-3

Cited by 36 publications

(32 citation statements)

References 24 publications

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“…The maximum of difference absorption for the active Pheo a was detected at 677.2 nm for Synechocystis (solid line) and 681.0 nm for spinach (dotted line) at 293 K; the extent of bleaching was comparable in the two preparations. The location of the active Pheo a has been reported at approximately 682 nm in PS II RC complexes of higher plants (Nanba and Satoh 1987;Braun et al 1990;Yruela et al 1994), consistent with this study. In contrast, the difference absorption spectrum of Synechocystis was blue-shifted by 3.8 nm, reflecting differences in the environment of Pheo a molecules.…”

Section: Pigment Compositionsupporting

confidence: 92%

“…This indicated that energy transfer occurred between Chl a and Pheo a. The fluorescence maxima of these spectra were consistent with previous reports from higher plants (Kwa et al 1992;Yruela et al 1994;Mimuro et al 1995). In the longer wavelength region of the maximum, the band shape was slightly broadened in Synechocystis; however, a corresponding long-wavelength component was not clear in the absorption spectrum (Fig.…”

Section: Fluorescence Spectrasupporting

confidence: 90%

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Isolation and spectral characterization of Photosystem II reaction center from Synechocystis sp. PCC 6803

et al. 2008

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We isolated highly-purified photochemically active photosystem (PS) II reaction center (RC) complexes from the cyanobacterium Synechocystis sp. PCC 6803 using a histidine-tag introduced to the 47 kDa chlorophyll protein, and characterized their spectroscopic properties. Purification was carried out in a one-step procedure after isolation of PS II core complex. The RC complexes consist of five polypeptides, the same as in spinach. The pigment contents per two molecules of pheophytin a were 5.8 +/- 0.3 chlorophyll (Chl) a and 1.8 +/- 0.1 beta-carotene; one cytochrome b(559) was found per 6.0 Chl a molecules. Overall absorption and fluorescence properties were very similar to those of spinach PS II RCs; our preparation retains the best properties so far isolated from cyanobacteria. However, a clear band-shift of pheophytin a and beta-carotene was observed. Reasons for these differences, and RC composition, are discussed on the basis of the three-dimensional structure of complexes.

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Section: Pigment Compositionsupporting

confidence: 92%

Section: Fluorescence Spectrasupporting

confidence: 90%

Isolation and spectral characterization of Photosystem II reaction center from Synechocystis sp. PCC 6803

et al. 2008

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“…D1-D2-Cyt b559 complex was purified according to the procedure of Nanba and Satoh (1987). However, Q-Sepharose Fast Flow (Pharmacia) was used instead of Fractogel DEAE-TSK 650s (Merck) as the ion-exchange chromatography column (2.6 x 21 cm) material, and MgS04 was substituted for NaCl as described by Yruela et al (1994). Specifically, the Q-Sepharose column was washed with 50 mM Tris-HC1, pH 7.2, 10 mM MgS04, plus 0.1% Triton X-100 at a flow rate of about 5 mL*min-' until the 435 nm peak of the eluate was a little lower than that at 416 nm.…”

Section: Preparation Of D L -D2-cyt B559 Complex and Pigmentmentioning

confidence: 99%

Pigment Content of D1-D2-Cytochrome b559 Reaction Center Preparations after Removal of CP47 Contamination: An Immunological Study

Pueyo¹,

Moliner²,

Seibert³

et al. 1995

Biochemistry

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Isolated D1 -D2-cytochrome b559 photosystem I1 reaction center preparations with pigment stoichiometry higher than 4 chlorophylls per 2 pheophytins can be contaminated with CP47 proximal antenna complex. Reaction centers prepared by a modification of the Nanba-Satoh procedure and containing about 6 chlorophylls per 2 pheophytins showed immuno-cross-reactivity when probed with a monoclonal antibody raised against the CP47 polypeptide. Furthermore, they could be fractionated successfully by Superose-12 sieve chromatography into two different populations. The first few fractions off the column contained a more definitive 435 nm shoulder corresponding to increased chlorophyll content, and showed strong immuno-cross-reactivity with the CP47 antibody. The peak fractions off the column displayed a less prominent 435 nm shoulder, and did not cross-react with the antibody. Moreover, when a 6-chlorophyll preparation was mixed with Sepharose beads coupled to CP47 antibody, the eluted material corresponded to a preparation of about 4 chlorophylls per 2 pheophytins and did not show any crossreaction with the antibody against CP47. The amount of CP47 protein in the 6-chlorophyll preparation as quantitated using Coomassie Blue staining or from gel blots was sufficient to account for most of the extra 2 chlorophylls. We conclude that D 1 -D2-cytochrome b559 preparations containing more than 4 chlorophylls per 2 pheophytins can be contaminated with small amounts of CP47-D1 -D2-Cyt b559 complex and that native photosystem I1 reaction centers contain 4 core chlorophylls per 2 pheophytins.

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“…PSII reaction center preparations (PSII-RC) contain only 4-6 Chl, 2 pheophytins (pheo), and 1-2 carotenoids (car) per PSII (17)(18)(19)(20)(21). Hence, destruction of less than one pigment per PSII can be easily observed in these particles (4,(22)(23)(24).…”

mentioning

confidence: 99%

Destruction of a single chlorophyll is correlated with the photoinhibition of photosystem II with a transiently inactive donor side.

Bumann

Oesterhelt

1995

Proc. Natl. Acad. Sci. U.S.A.

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Pigments destroyed during photoinhibition of water-splitting photosystem II core complexes from the green alga Chiamydomonas reinhardtii were studied. Under conditions of a transiently inactivated donor side, illumination leads to an irreversible inhibition of the electron transfer at the donor side that is paralleled by the destruction of chlorophylls a absorbing maximally around 674 and 682 nm. The observed stochiometry of 1 + 0.1 destroyed chlorophyll per inhibited photosystem II suggests that chlorophyll destruction could be the primary photodamage causing the inhibition of photosystem II under these conditions.In photosynthesis light energy is converted to chemical energy. However, side reactions can lead to considerable destruction of the photosynthetic apparatus and a concomitant loss of photosynthetic activity in a process called photoinhibition (1). In oxygenic photosynthesis the main target for photoinhibition is photosystem II (PSII) (2). In intact PSII the excited primary donor P680 reduces plastoquinone. The oxidized primary donor P680+ is then reduced by water via an redoxactive tyrosine "Z" (3). In case of a transient malfunction of the water-splitting reaction, P680+ has an extended lifetime during which it can degrade (4) or damage other PSII components, including carotenoids, chlorophylls (Chl), possibly Z, the manganese binding sites, and other amino acids of the PSII proteins (5-10). One or several of these damages result in an irreversible inhibition of the electron transfer from Z to P680+ and hence in an irreversible loss of the water-splitting activity (6,7,(10)(11)(12)(13)(14). Despite many investigations it is still not clear which of the various damages actually causes this inhibition. We investigated this problem by correlating pigment destruction with the loss of oxygen evolving activity.Quantitative observations of pigment destruction in vivo or in grana membranes are difficult due to the high number of pigments per PSII (200-700) (15, 16). PSII reaction center preparations (PSII-RC) contain only 4-6 Chl, 2 pheophytins (pheo), and 1-2 carotenoids (car) per PSII (17-21). Hence, destruction of less than one pigment per PSII can be easily observed in these particles (4,(22)(23)(24). However, since the electron transfer from Z to P680+ is already strongly impaired in freshly isolated PSII-RC (25-27), these particles cannot be used for the investigation of the first irreversibly inactivating reaction of photoinhibition. We therefore used PSII core complexes, which have an intermediate pigment content and are capable of water splitting (28). With these particles it is possible to observe the destruction of less than one Chl per PSII and to measure the loss of water-splitting activity (including the electron transfer from Z to P680+) in parallel.The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact. Experimental ProceduresWater-s...

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Characterization of a D1‐D2‐cyt b‐559 complex containing 4 chlorophyll a/2 pheophytin a isolated with the use of MgSO₄

Cited by 36 publications

References 24 publications

Isolation and spectral characterization of Photosystem II reaction center from Synechocystis sp. PCC 6803

Isolation and spectral characterization of Photosystem II reaction center from Synechocystis sp. PCC 6803

Pigment Content of D1-D2-Cytochrome b559 Reaction Center Preparations after Removal of CP47 Contamination: An Immunological Study

Destruction of a single chlorophyll is correlated with the photoinhibition of photosystem II with a transiently inactive donor side.

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