Isolation and characterization of an oxygen-evolving Photosystem II reaction center core preparation and a 28 kDa Chl-a-binding protein

Ghanotakis, Demetrios F.; Demetriou, D. M.; Yocum, Charles F.

doi:10.1016/0005-2728(87)90078-8

Cited by 249 publications

(139 citation statements)

References 36 publications

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“…In contrast, Bassi et al [5] suggested a close association of a complex between CP26 and CP29 to CP43 of the reaction-centre core of PS 11, via a so-called CP24 linker. The results of Ghanotakis et al [34] and Camm et al [29] likewise supported the proposal that CP29 belongs to the PS-I1 core complex [35]. This is in line with our density-gradient experiments after short solubilization conditions (Fig.…”

Section: Bi~diiirlg Of'bivc~lr~it Cutiorrs To the Pigmelit-protein Cosupporting

confidence: 92%

Isolation, purification and partial characterization of a 30‐kDa chlorophyll‐a/b‐binding protein from spinach

Irrgang¹,

Ренгер²,

Vater

1991

European Journal of Biochemistry

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A 30-kDa chlorophyll-a/b-binding protein was purified from photosystem I1 membrane fragments using Ca2+-chelating Sepharose 6B chromatography. The protein binds approximately four chlorophyll a molecules, one chlorophyll b molecule and carotenoids. Its 77-K fluorescence-emission spectrum exhibits a maximum at 680 k 1 nm. The protein has a high tendency to form a dimer in the presence of Ca2+. Ca2+ binding affects the low-temperature fluorescence-emission maximum, leading to a decrease in its intensity and a blue shift of 1 nm. Similar spectral changes were obtained in the presence of Mg2+, possibly indicating a common binding domain for both cations. We interpret these observations as cation-induced conformational changes of the protein, which were reversible upon subsequent incubation in EDTA. Evidence is presented for the involvement of carboxyl groups in the coordination sphere of the bivalent cations. The possible structural and functional role of the protein is discussed.The primary physicochemical events of photosynthesis comprise light absorption, excitation, energy transfer and exciton transformation into electrochemical free energy by the formation of a sufficiently stable radical pair within a functional unit, referred to as the reaction-centre complex. In higher plants, different types of chlorophyll (Ch1)-containing protein complexes are incorporated into the thylakoid membrane: Chl-a/b-containing peripheral antenna complexes exhibiting lateral mobility; immobile core antenna complexes containing either Chl a/b or Chl a and reaction-centre complexes of photosystem I and 11, respectively (for recent reviews see [I -31). The mobile peripheral light-harvesting antenna complex of PS I1 (LHC 11) carries about 120 Chl a/ h molecules bound to at least two distinct polypeptides with relative molecular masses of 25 kDa and 27 kDa [I]. The immobile antenna is composed of about 80 k 20 Chl a/b molecules bound to polypeptides of 24 (CP24), 26 (CP26) and 29 (CP29) kDa [I -51. The pigment distribution among these polypeptides is a matter of debate. In addition, two Chl-acontaining proteins of 43 kDa and 47 kDa are closely associated with the PS I1 reaction-centre complex. Each of these polypeptides is assumed to bind about 25 Chl a molecules [6, 71. Both proteins presumably also function as core antennae, but for CP47 in particular an additional role in maintaining the structural integrity of the reaction-centre complex is not excluded [5,8,[9][10][11][12].The isolation and characterization of Chl-binding proteins is difficult to achieve with commonly used protein-chemistry Abbreviations. Chl, chlorophyll; CP, chlorophyll-protein complex; LHC, light-harvesting complex; P680, reaction-centre chlorophyll u molccule of photosystem 11; PS I and PS 11, photosystems I and 11.techniques because of the tendency of such proteins to selfassociate into oligomers, their similarities in relative molecular masses, isoelectric points and hydrophobicities and structural homologies. Many of these polypeptides have therefore been p...

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Section: Bi~diiirlg Of'bivc~lr~it Cutiorrs To the Pigmelit-protein Cosupporting

confidence: 92%

Isolation, purification and partial characterization of a 30‐kDa chlorophyll‐a/b‐binding protein from spinach

Irrgang¹,

Ренгер²,

Vater

1991

European Journal of Biochemistry

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“…A single peak was found ( suggesting that the preparation consists of a homogneous population of particles. Tris-washed, solubilized reaction center complexes show a second peak (Figure 1, solid line) which is due primarily to monomeric 43-and 28-kDa polypeptides and is not observed in the isolated CP47-Dl-D2-b-559 complex [see Ghanotakis et al (1987) for a similar experiment using oxygen-evolving fractions]. Comparison of these data with elution patterns of standard proteins produced an estimated molecular mass of 200-300 kDa for the CP47-Dl-D2-b-559 material.…”

Section: Resultsmentioning

confidence: 99%

“…Subchloroplast PS I1 membranes and oxygen-evolving PS I1 reaction center complexes were prepared from spinach (Ghanotakis et al, 1987), and CP47-Dl-D2-b-559 complexes were prepared as described by Dekker et al (1989). For crystallization, the final fraction in buffer A (20 mM Bis-Tris, 20 mM NaC1, 10 mM MgCI2, 1.5% taurine, and 0.03% dodecyl maltoside, pH 6.5) with 100 mM MgS04 was concen- trated slowly by dialysis against solid sucrose.…”

Section: Methodsmentioning

confidence: 99%

Characterization by electron microscopy of isolated particles and two-dimensional crystals of the CP47-D1-D2-cytochrome b-559 complex of photosystem II

Dekker¹,

Betts²,

Yocum³

et al. 1990

Biochemistry

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A photosystem II complex containing the reaction center proteins Dl and D2, a 47-kDa chlorophyll-binding protein (CP47), and cytochrome 6-559 was isolated with high yield, purity, and homogeneity; small but well-ordered two-dimensional crystals were prepared from the particles. The crystals and the isolated particles were analyzed by electron microscopy using negatively stained specimens. The t This research was supported by National Science Foundation Grant DCB 89-04075 to C.F.Y. and by grants from the Deutsche Forschungs Gemeinschaft, Sonderforschungsbereich 312, to E.J.B.

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“…Photosystem II membranes or BBY particles were isolated from thylakoid membranes as described by (Berthold et al 1981) with some modifications as described by (Van Leeuwen et al 1991) and dissolved in the same buffer as the native membranes. Additionally GY particles (named after Ghanotakis and Yocum who where the first ones to isolate this particle), which consist of the PSII core complex and the minor antenna complexes CP29 and CP26 were prepared as in Ghanotakis et al (1987). The PSI-LHCI complexes were isolated from unstacked spinach thylakoid membranes as described in Dekker et al (2002) and diluted in a buffer of 20 mM BisTris pH 6.5, 5 mM MgCl 2 and 0.06% of b-DM.…”

Section: Methodsmentioning

confidence: 99%

“…2. For an estimation of the sum of the first two contributions, fluorescence excitation spectra of GY membranes were used, which consist of the PSII core complex and the minor antenna complexes CP29 and CP26 (Ghanotakis et al 1987). The GY excitation spectrum accounts for absorption strength in the red flank of the excitation difference spectrum (caused by red Chls in the core antenna proteins CP47 and CP43) and has a higher Chl a/b ratio than LHCII.…”

Section: K Fluorescence Excitationmentioning

confidence: 99%

Excitation energy transfer in native and unstacked thylakoid membranes studied by low temperature and ultrafast fluorescence spectroscopy

et al. 2007

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In this work, the transfer of excitation energy was studied in native and cation-depletion induced, unstacked thylakoid membranes of spinach by steady-state and time-resolved fluorescence spectroscopy. Fluorescence emission spectra at 5 K show an increase in photosystem I (PSI) emission upon unstacking, which suggests an increase of its antenna size. Fluorescence excitation measurements at 77 K indicate that the increase of PSI emission upon unstacking is caused both by a direct spillover from the photosystem II (PSII) core antenna and by a functional association of light-harvesting complex II (LHCII) to PSI, which is most likely caused by the formation of LHCII-LHCI-PSI supercomplexes. Time-resolved fluorescence measurements, both at room temperature and at 77 K, reveal differences in the fluorescence decay kinetics of stacked and unstacked membranes. Energy transfer between LHCII and PSI is observed to take place within 25 ps at room temperature and within 38 ps at 77 K, consistent with the formation of LHCII-LHCI-PSI supercomplexes. At the 150-160 ps timescale, both energy transfer from LHCII to PSI as well as spillover from the core antenna of PSII to PSI is shown to occur at 77 K. At room temperature the spillover and energy transfer to PSI is less clear at the 150 ps timescale, because these processes compete with charge separation in the PSII reaction center, which also takes place at a timescale of about 150 ps.

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Isolation and characterization of an oxygen-evolving Photosystem II reaction center core preparation and a 28 kDa Chl-a-binding protein

Cited by 249 publications

References 36 publications

Isolation, purification and partial characterization of a 30‐kDa chlorophyll‐a/b‐binding protein from spinach

Isolation, purification and partial characterization of a 30‐kDa chlorophyll‐a/b‐binding protein from spinach

Characterization by electron microscopy of isolated particles and two-dimensional crystals of the CP47-D1-D2-cytochrome b-559 complex of photosystem II

Excitation energy transfer in native and unstacked thylakoid membranes studied by low temperature and ultrafast fluorescence spectroscopy

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