Crystallization and preliminary crystallographic characterization of the extrinsic PsbP protein of photosystem II from<i>Spinacia oleracea</i>

Kohoutová, Jaroslava; Smatanová, Ivana Kutá; Brynda, J.; Lapkouski, Mikalai; Revuelta, José Luis; Arellano, Juan B.; Ettrich, Rüdiger

doi:10.1107/s1744309108040578

Cited by 13 publications

(8 citation statements)

References 15 publications

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“…They may also impact the association of Ca 2+ and Cl 2 with PSII, the polypeptide conformation around the manganese cluster, and the formation of channels within PSII that allow access of water to the catalytic site and the exit of protons from the complex as water is oxidized (Bricker et al, 2012). The x-ray crystal structures of purified PsbP (Kohoutová et al, 2009) and PsbQ (Balsera et al, 2005) from spinach (Spinacia oleracea) have been determined. For cyanobacteria, PSII crystals have been used to establish high-resolution structures for PsbO, PsbV, and PsbU (Umena et al, 2011), with more recent analyses at room temperature (Kern et al, 2013).…”

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confidence: 99%

The Use of Contact Mode Atomic Force Microscopy in Aqueous Medium for Structural Analysis of Spinach Photosynthetic Complexes

et al. 2015

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To investigate the dynamics of photosynthetic pigment-protein complexes in vascular plants at high resolution in an aqueous environment, membrane-protruding oxygen-evolving complexes (OECs) associated with photosystem II (PSII) on spinach (Spinacia oleracea) grana membranes were examined using contact mode atomic force microscopy. This study represents, to our knowledge, the first use of atomic force microscopy to distinguish the putative large extrinsic loop of Photosystem II CP47 reaction center protein (CP47) from the putative oxygen-evolving enhancer proteins 1, 2, and 3 (PsbO, PsbP, and PsbQ) and large extrinsic loop of Photosystem II CP43 reaction center protein (CP43) in the PSII-OEC extrinsic domains of grana membranes under conditions resulting in the disordered arrangement of PSII-OEC particles. Moreover, we observed uncharacterized membrane particles that, based on their physical characteristics and electrophoretic analysis of the polypeptides associated with the grana samples, are hypothesized to be a domain of photosystem I that protrudes from the stromal face of single thylakoid bilayers. Our results are interpreted in the context of the results of others that were obtained using cryo-electron microscopy (and single particle analysis), negative staining and freeze-fracture electron microscopy, as well as previous atomic force microscopy studies.

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confidence: 99%

The Use of Contact Mode Atomic Force Microscopy in Aqueous Medium for Structural Analysis of Spinach Photosynthetic Complexes

et al. 2015

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“…The crystal structure of spinach PsbP (PDB: 2VU4) has been recently reported (24). Interestingly, the structure of spinach PsbP has one Zn 2ϩ ion that is ligated to the Asp-165 and His-144 residues in the C-terminal domain.…”

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confidence: 99%

The Conserved His-144 in the PsbP Protein Is Important for the Interaction between the PsbP N-terminus and the Cyt b559 Subunit of Photosystem II

Ido

Kakiuchi

Uno

et al. 2012

Journal of Biological Chemistry

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Background:PsbP is an extrinsic subunit of photosystem II in green plants. Results: H144A mutation in PsbP alters the chloride requirement and affects the interaction between its N terminus and the PsbE component of photosystem II. Conclusion:The N-and C-terminal domains of PsbP cooperate to support PSII activity. Significance: This provides important information about the binding characteristics of PsbP in green plant PSII.The PsbP protein regulates the binding properties of Ca 2؉ and Cl ؊ , and stabilizes the Mn cluster of photosystem II (PSII); however, the binding site and topology in PSII have yet to be clarified. Here we report that the structure around His-144 and Asp-165 in PsbP, which is suggested to be a metal binding site, has a crucial role for the functional interaction between PsbP and PSII. The mutated PsbP-H144A protein exhibits reduced ability to retain Cl ؊ anions in PSII, whereas the D165V mutation does not affect PsbP function. Interestingly, H144A/D165V double mutation suppresses the effect of H144A mutation, suggesting that these residues have a role other than metal binding. FTIR difference spectroscopy suggests that H144A/D165V restores proper interaction with PSII and induces the conformational change around the Mn cluster during the S 1 /S 2 transition. Cross-linking experiments show that the H144A mutation affects the direct interaction between PsbP and the Cyt b 559 ␣ subunit of PSII (the PsbE protein). However, this interaction is restored in the H144A/D165V mutant. In the PsbP structure, His-144 and Asp-165 form a salt bridge. H144A mutation is likely to disrupt this bridge and liberate Asp-165, inhibiting the proper PsbP-PSII interaction. Finally, mass spectrometric analysis has identified the cross-linked sites of PsbP and PsbE as Ala-1 and Glu-57, respectively. Therefore His-144, in the C-terminal domain of PsbP, plays a crucial role in maintaining proper N terminus interaction. These data provide important information about the binding characteristics of PsbP in green plant PSII.Photosystem II (PSII) 2 consists of both membrane-intrinsic and membrane-extrinsic subunits, and functions as a water/ plastoquinone oxidoreductase (for reviews, Refs. 1-4). On the thylakoid lumenal side of PSII, a metal cluster of four Mn ions, one Ca 2ϩ ion, and five oxo ligands (the Mn cluster) catalyzes the oxygen-evolving reaction. Additionally, two Cl Ϫ ions are bound near to the Mn cluster (5). The membrane-intrinsic subunits of PSII are involved in pigment and/or cofactor binding for photochemical reactions, while the membrane-extrinsic subunits surround the catalytic Mn cluster and play crucial roles in stabilizing the Mn cluster and retaining the PSII cofactor ions (6 -8).X-ray structural analysis of the cyanobacterial PSII complex at atomic resolution has revealed the location of subunits, pigments, and cofactors, including the exact organization of the subunits within PSII (5, 9 -11). However, the crystallographic information gained from the study of cyanobacterial PSII cannot necessarily be...

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“…The expression protocol followed Kohoutová et al ( 2009 ) with some modifications. The gene coding for PsbP was included in the JR3133 plasmid, which also includes a His 6 tag attached to the N-terminus of the protein.…”

Section: Methods and Experimentsmentioning

confidence: 99%

“…The three dimensional structure of PsbP from Spinacia oleracea has been resolved recently by X-ray crystallography at high resolution (1.98 Å) (Kopecký et al 2012 ; Kohoutová et al 2009 ), see Fig. 1 .…”

Section: Biological Contextmentioning

confidence: 99%

Resonance assignment of PsbP: an extrinsic protein from photosystem II of Spinacia oleracea

et al. 2015

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PsbP (23 kDa) is an extrinsic eukaryotic protein of photosystem II found in the thylakoid membrane of higher plants and green algae. It has been proven to be indispensable for proper functioning of the oxygen evolving complex. By interaction with other extrinsic proteins (PsbQ, PsbO and PsbR), it modulates the concentration of two cofactors of the water splitting reaction, Ca2+ and Cl−. The crystallographic structure of PsbP from Spinacia oleracea lacks the N-terminal part as well as two inner regions which were modelled as loops. Those unresolved parts are believed to be functionally crucial for the binding of PsbP to the thylakoid membrane. In this NMR study we report 1H, 15N and 13C resonance assignments of the backbone and side chain atoms of the PsbP protein. Based on these data, an estimate of the secondary structure has been made. The structural motifs found fit the resolved parts of the crystallographic structure very well. In addition, the complete assignment set provides preliminary insight into the dynamic regions.

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Crystallization and preliminary crystallographic characterization of the extrinsic PsbP protein of photosystem II fromSpinacia oleracea

Cited by 13 publications

References 15 publications

The Use of Contact Mode Atomic Force Microscopy in Aqueous Medium for Structural Analysis of Spinach Photosynthetic Complexes

The Use of Contact Mode Atomic Force Microscopy in Aqueous Medium for Structural Analysis of Spinach Photosynthetic Complexes

The Conserved His-144 in the PsbP Protein Is Important for the Interaction between the PsbP N-terminus and the Cyt b559 Subunit of Photosystem II

Resonance assignment of PsbP: an extrinsic protein from photosystem II of Spinacia oleracea

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