Pigment stoichiometry of the Photosystem II reaction center from higher plants

Aured, Maria; Moliner, Esmeralda; Alfonso, Miguel; Yruela, Inmaculada; Toon, S.; Seibert, Michael; Picorel, Rafael

doi:10.1016/0005-2728(94)90108-2

Cited by 13 publications

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“…Alfonso et al, 1994), respectively. Quantitation by the Coomassie Blue-staining method probably overestimates the amount of CP47 present since CP47 stains more heavily than D1 and D2, even though we have corrected for the higher molecular weight of CP47.…”

Section: Resultsmentioning

confidence: 99%

“…This method of purification (Aured et al, 1994) leads to the fractionation of the initial PSII RC population (6 Chl per 2 Pheo; Figure 1A) into two components without significant pigment loss. The first component contains RC complex with more than 6 Chl per 2 Pheo (Figure lC), and the main component (sieve chromatography peak) contains RC complex with about 4 Chl per 2 Pheo ( Figure 1B).…”

Section: Resultsmentioning

confidence: 99%

“…An absorption spectrum of each fraction was taken. The few first fractions (' 6 Chl per 2 Pheo), which contained a pronounced shoulder at 435 nm, were pooled as a component set, and the main peak (4 Chl per 2 Pheo) was collected as another component set (Aured et al, 1994). Sometimes, complete separation was not achieved (Le., RC preparations with 4.5-5 Chl per 2 Pheo have been observed).…”

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

confidence: 99%

“…For Table 1, CP47 protein was purified as described previously (Alfonso et al, 1994). The amount of the purified protein was determined from the spectrum of the pigment extract in 80% acetone, using an extinction coefficient of 75 mM-l cm-' at 663 nm for chlorophyll (Dawson et al, 1986).…”

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

confidence: 99%

“…Since significant sequence homology between the L and M polypeptides of the purple bacterial RC and the D1 and D2 polypeptides of the PSII RC exists in the regions thought to bind pigments (Youvan et al, 1984;Rochaix et al, 1984), the core Chl pigment content in the two types of RC could be similar. Recent reports suggest the presence of pigment-protein and or free Chl contaminants in preparations of isolated D1-D2-Cyt b559 RC complex containing more than 4 Chl per 2 Pheo (Jankowiak & Small, 1993;Aured et al, 1994;Chang et al, 1994a). Resolving this issue is important since the exact chromophore stoichiometry is critical for understanding functional mechanisms in the PSII RC including excited state phenomena, energy transfer, and primary charge separation.…”

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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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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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

confidence: 99%

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

confidence: 99%

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confidence: 99%

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