Localization of the PsbH subunit in photosystem II from the Synechocystis 6803 using the His-tagged Ni–NTA Nanogold labeling

Bumba, Ladislav; Tichý, Martin; Dobáková, Marika; Komenda, Josef; Vacha, František

doi:10.1016/j.jsb.2005.08.001

Cited by 19 publications

(15 citation statements)

References 35 publications

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“…Electron microscopy and single particle analysis did not reveal any differences between PSII dimers isolated from normal or HL-treated cells, indicating that SCPs do not form large complexes at or around PSII comparable to IsiA rings formed around the PSI trimers in iron-depleted conditions (57). When compared with the location of the PSII subunit PsbH using a similar approach (52), localization of the gold label on the ScpDHis protein is less distinct (Fig. 7, D and E).…”

Section: Discussionmentioning

confidence: 91%

“…This is probably not the case as the native forms of ScpD and ScpC seem to be also associated with PSII (see below). We also attempted to isolate complexes containing ScpDHis under native conditions by nickel affinity chromatography in a similar procedure that we have used recently for the His-tagged PsbH protein (52). However, no ScpDHis protein was found in the fraction eluted from the column.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, a different labeling procedure than the one used recently for the His-tagged PsbH protein (52) had to be developed for a successful labeling. The combination of Ni-NTA Nanogold labeling of His-tagged protein and single particle analysis provides an excellent tool to localize protein subunits within a protein complex (52,56). The main advantage to conventional immunogold labeling procedures is the close proximity of the gold particle to the His tag, enabling a more accurate localization of the target protein.…”

Section: Discussionmentioning

confidence: 99%

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Cyanobacterial Small Chlorophyll-binding Protein ScpD (HliB) Is Located on the Periphery of Photosystem II in the Vicinity of PsbH and CP47 Subunits

Promnares

Komenda

Bumba

et al. 2006

Journal of Biological Chemistry

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Cyanobacteria contain several genes coding for small one-helix proteins called SCPs or HLIPs with significant sequence similarity to chlorophyll a/b-binding proteins. To localize one of these proteins, ScpD, in the cells of the cyanobacterium Synechocystis sp. PCC 6803, we constructed several mutants in which ScpD was expressed as a His-tagged protein (ScpDHis). Using two-dimensional native-SDS electrophoresis of thylakoid membranes or isolated Photosystem II (PSII), we determined that after high-light treatment most of the ScpDHis protein in a cell is associated with PSII. The ScpDHis protein was present in both monomeric and dimeric PSII core complexes and also in the core subcomplex lacking CP43. However, the association with PSII was abolished in the mutant lacking the PSII subunit PsbH. In a PSII mutant lacking cytochrome b 559 , which does not accumulate PSII, ScpDHis is associated with CP47. The interaction of ScpDHis with PsbH and CP47 was further confirmed by electron microscopy of PSII labeled with Ni-NTA Nanogold. Single particle image analysis identified the location of the labeled ScpDHis at the periphery of the PSII core complex in the vicinity of the PsbH and CP47. Because of the fact that ScpDHis did not form any large structures bound to PSII and because of its accumulation in PSII subcomplexes containing CP47 and PsbH we suggest that ScpD is involved in a process of PSII assembly/repair during the turnover of pigment-binding proteins, particularly CP47.

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Section: Discussionmentioning

confidence: 91%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Cyanobacterial Small Chlorophyll-binding Protein ScpD (HliB) Is Located on the Periphery of Photosystem II in the Vicinity of PsbH and CP47 Subunits

Promnares

Komenda

Bumba

et al. 2006

Journal of Biological Chemistry

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“…For specific labeling, Au NPs are functionalized with certain ligands that only bind to the target sequence in a protein, and the location of the specific subunit can be determined by TEM observations. [1][2][3][4] Compared with the antibody labeling method, Au NP labeling has a higher resolution. To obtain a stable signal and achieve a better resolution, wellfunctionalized Au NPs with the appropriate size and size distribution are desired.…”

Section: Introductionmentioning

confidence: 99%

Simple Method of Synthesizing Nickel–Nitrilotriacetic Acid Gold Nanoparticles with a Narrow Size Distribution for Protein Labeling

Kitai

Watanabe

Toyoshima

et al. 2011

Jpn. J. Appl. Phys.

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We developed a simple method to synthesize nickel–nitrilotriacetic acid gold nanoparticles (Ni–NTA Au NPs) with a narrow size distribution for site-specific labeling in protein complexes. Au NPs were synthesized by the reduction of HAuCl4 using trisodium citrate and tannin acid. Then, the nanoparticle surfaces were modified with NTA and subsequent complexation with Ni2+. The mean diameter of the synthesized Ni–NTA Au NPs was 4.3 nm, and the coefficient of variation was 9%. The specific binding of the Ni–NTA Au NPs to polyhistidine-tagged (His-tagged) proteins was determined by transmission electron microscopy using kinesin and the p62 subunit of dynactin. Consequently, our method is useful for analyzing the substructures of protein complexes.

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“…The redox characteristics exhibited in the spectra are unlikely to arise from the 22 and 10 kDa proteins, which are encoded by the psbS and psbH genes respectively, because they are not known to be redox active. The PsbS protein has been demonstrated to control the organization of the PS 2 antenna in higher plant thylakoid membranes (Kiss et al 2008) whereas the PsbH protein belongs to a group of small protein subunits of PS 2 and thought to be involved in the assembly of the PS 2 complex (Bumba et al 2005). The 22 and 10 kDa PS 2 proteins, which have also been purified to homogeneity from PS 2 complex, do not absorb in the visible spectral range (Ljungberg et al 1986).…”

Section: ⎯⎯⎯⎯mentioning

confidence: 99%

Simple method for isolation of oxygen evolving photosystem 2 core complex free of cytochrome f contamination

Mishra¹

2010

Biologia plant.

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This communication presents a simple method for isolation of oxygen evolving photosystem 2 (PS 2) core complex by solubilisation of PS 2 membranes with the nonionic detergent octyl-β-D-thioglucopyranoside (OTG). This complex was free of cytochrome (Cyt) f contamination and also lacked the 22 and 10 kDa proteins that may not be directly required for primary photochemistry of the PS 2 complex and water oxidation. OTG could also remove the Cyt f contamination from the PS 2-core complex isolated using octyl-β-D-glucopyranoside (OGP). The Cyt f contamination in the PS 2 complexes could potentially interfere with spectrophotometric determinations of redox states of Cytb 559 and its stoichiometry in the PS 2 complex.Additional key words: octyl-β-D-thioglucopyranoside, oxygen evolution, spinach.

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Localization of the PsbH subunit in photosystem II from the Synechocystis 6803 using the His-tagged Ni–NTA Nanogold labeling

Cited by 19 publications

References 35 publications

Cyanobacterial Small Chlorophyll-binding Protein ScpD (HliB) Is Located on the Periphery of Photosystem II in the Vicinity of PsbH and CP47 Subunits

Cyanobacterial Small Chlorophyll-binding Protein ScpD (HliB) Is Located on the Periphery of Photosystem II in the Vicinity of PsbH and CP47 Subunits

Simple Method of Synthesizing Nickel–Nitrilotriacetic Acid Gold Nanoparticles with a Narrow Size Distribution for Protein Labeling

Simple method for isolation of oxygen evolving photosystem 2 core complex free of cytochrome f contamination

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