Site-Directed Mutagenesis of the CP 47 Protein of Photosystem II:  Alteration of Conserved Charged Residues in the Domain <sup>364</sup>E−<sup>444</sup>R

Putnam-Evans, Cindy; Burnap, Robert L.; Wu, Jituo; Whitmarsh, John; Bricker, Terry M.

doi:10.1021/bi952661s

Cited by 47 publications

(51 citation statements)

References 36 publications

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“…PsbO is positioned over the CP47 side of the reaction center (Zouni et al 2001). This is in agreement with experiments showing that deletions in the E loops of CP47 combined with the deletion of PsbO abolished photoautotrophic growth (Morgan et al 1998;Clarke and Eaton-Rye 1999), and site directed mutagenesis studies that show a binding domain for PsbO at CP47 Arg384 and Arg385 (PutnamEvans and Bricker 1992;Putnam-Evans et al 1996;Qian et al 1997). Additionally, PsbO stabilizes the AB loop and C-terminus of D1 (the location of many of the manganese cluster ligands), and interacts with the large E loop of CP47 (Ferreira et al 2004;Nield and Barber 2006).…”

Section: Cyanobacterial Oecsupporting

confidence: 84%

The extrinsic proteins of Photosystem II

2007

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Years of genetic, biochemical, and structural work have provided a number of insights into the oxygen evolving complex (OEC) of Photosystem II (PSII) for a variety of photosynthetic organisms. However, questions still remain about the functions and interactions among the various subunits that make up the OEC. After a brief introduction to the individual subunits Psb27, PsbP, PsbQ, PsbR, PsbU, and PsbV, a current picture of the OEC as a whole in cyanobacteria, red algae, green algae, and higher plants will be presented. Additionally, the role that these proteins play in the dynamic life cycle of PSII will be discussed.

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Section: Cyanobacterial Oecsupporting

confidence: 84%

The extrinsic proteins of Photosystem II

2007

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“…Deletion of short parts of CP47 between residues 373 and 392 in Synechocystis PCC 6803 results in a weakened binding of the 33-kDa protein. Deletion of residues 416-420 (Gleiter et al, 1994), which includes lysines 41 8 and 419, or site-directed mutagenesis of the latter two residues (Putnam-Evans et al, 1996) did not change the binding of the 33-kDa protein to PSII to a significant extent. Therefore, it is likely that K389 is the residue involved in cross-linking.…”

Section: Discussionmentioning

confidence: 84%

Intermolecular and Intramolecular Interactions of the 33‐kDa Protein in Photosystem II

Seidler

1996

European Journal of Biochemistry

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In this work, cross-linking was observed not only to CP47 but also to a small intrinsic subunit. In addition, through the use of a high-resolution SDS-gel system, three intramolecular cross-linked products of the 33-kDa protein were detected. To search for additional cross-linking sites that might not be accessible to the cross-linker in intact photosystem 11, the isolated 33-kDa protein was activated for cross-linking and subsequently bound to CaC1,-washed photosystem 11. In the complementary experiment, CaC1,-washed photosystem I1 was activated, then reconstituted with the 33-kDa protein. The results of the crosslinking reactions demonstrated that all carboxylic acid groups involved in cross-linking were located on the 33-kDa protein and all primary amines were located on intrinsic membrane proteins. No cross-linking other than those observed in cross-linking experiments with intact photosystem I1 were observed. This indicated that the 33-kDa protein is bound to CP47 and a small subunit but not to the photosystem I1 reaction centre. This observation is consistent with the finding that cross-linking was independent of the presence or absence of the manganese cluster. Possible residues on the 33-kDa protein and CP47 involved in cross-linking are suggested.Keywords: photosystem 11; oxygen evolution; cross-linking ; l-ethyl-3-(3-dimethylaminopropyl)carbodiimide ; protein-protein interaction.Three extrinsic polypeptides that are involved in oxygen evolution are located on the lumenal side of photosystem I1 (PSII) from higher plants. These 33-kDa, 23-kDa and 16-kDa proteins have been extensively studied by dissociation and reconstitution experiments (reviewed by Seidler, 1996). They can be removed by treatment with 1 M Tris, pH 8.0, 2.6 M urea, or with alkaline solutions with concomitant loss of the manganese cluster, the catalytic centre of oxygen evolution. NaCl treatment of PSII removes selectively the 23-kDa and 16-kDa polypeptides. However, the function of these proteins can be replaced by 5 mM Caz+ and 30 mM C1-. Treatment of PSI1 with 1 M CaCI, Inoue, 1983, 1984) or 2.6 M urea and 200 mM NaCl Murata, 1983, 1984) releases all three extrinsic proteins without significant loss of manganese. The activity of PSI1 is lowered to 16-40%, even in the presence of optimal concentrations of Ca2+ and C1-, and the manganese cluster becomes extremely sensitive to light, elevated temperature and endogenous reductants. Rebinding of the 33-kDa protein restores oxygen-evolving activity and the stability of the manganese complex.Despite intensive research, there is still some controversy about where the three extrinsic polypeptides are bound. This question is particularly important for the 33-kDa protein because it appears to be the extrinsic protein which is the most important for oxygen evolution. The strongest evidence has been found for binding of the 33-kDa protein to the chlorophyll-a-binding protein CP47. Removal of the 33-kDa protein was necessary to bind a lysine modifier, N-hydroxysuccinimide-activated biotin to the...

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“…Mutants containing alterations in other PS II components that cannot bind the PsbO protein normally exhibit similar characteristics (Ref. 21; for an in-depth review, see Ref. 22).…”

Section: Resultsmentioning

confidence: 99%

The psbo1 Mutant of Arabidopsis Cannot Efficiently Use Calcium in Support of Oxygen Evolution by Photosystem II

Bricker

Frankel

2008

Journal of Biological Chemistry

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The Arabidopsis thaliana mutant psbo1 contains a point mutation in the psbO-1 gene (At5g66570) leading to the loss of expression of the PsbO-1 protein and overexpression of the PsbO-2 protein (Murakami, R., Ifuku, K., Takabayashi, A., Shikanai, T., Endo, T., and Sato, F. (2002) FEBS Lett. 523, 138 -142). Previous characterization of fluorescence induction and decay kinetics by our laboratory documented defects on both the oxidizing and reducing sides of Photosystem II. Additionally, anomalous flash oxygen yield patterns indicated that the mutant contains a defective oxygen-evolving complex that appears to exhibit anomalously long-lived S 2 and S 3 oxidation states (Liu, H., Frankel, L. K., and Bricker, T. M. (2007) Biochemistry 46, 7607-7613). In this study, we have documented that the S 2 and S 3 states in psbo1 thylakoids decay very slowly. The total flash oxygen yield of the psbo1 mutant was also significantly reduced, as was its stability. Incubation of psbo1 thylakoids at high NaCl concentrations did not increase the rate of S 2 and S 3 state decay. The oxygen-evolving complexes of the mutant did, however, exhibit somewhat enhanced stability following this treatment. Incubation with CaCl 2 had a significantly more dramatic effect. Under this condition, both the S 2 and S 3 states of the mutant decayed at nearly the same rate as the wild type, and the total oxygen yield and its stability following CaCl 2 treatment were indistinguishable from that of the wild type. These results strongly suggest that the principal defect in the psbo1 mutant is an inability to effectively utilize the calcium associated with Photosystem II. We hypothesize that the PsbO-2 protein cannot effectively sequester calcium at the oxygen-evolving site. Photosystem II (PS II)2 functions as a light-driven, waterplastoquinone oxidoreductase. In higher plants and cyanobacteria at least six intrinsic proteins appear to be required for O 2 evolution (1-3). These are CP47, CP43, D1, D2, and the ␣ and ␤ subunits of cytochrome b 559 . Deletion of these subunits uniformly results in the loss of PS II function and assembly (4). Additionally, in higher plants, three extrinsic proteins, with apparent molecular masses of 33 kDa (PsbO), 24 kDa (PsbP), and 16 kDa (PsbQ), are also required for maximal rates of O 2 evolution at physiological inorganic cofactor concentrations. Of these three proteins, the PsbO protein appears to play a central role in the stabilization of the manganese cluster, is essential for efficient and stable O 2 evolution, and is required, along with PsbP, for photoautotrophic growth and PS II assembly in higher plants propagated under normal growth conditions (5, 6).In Arabidopsis thaliana, two genes that encode PsbO (psbO-1, At5g66570 and psbO-2, At3g50820) are normally expressed, yielding two different PsbO proteins (PsbO-1 and PsbO-2, respectively). A highly fluorescent mutant, psbo1, was recently identified in which a stop codon has been introduced in the psbO-1 gene by ethane methylsulfonate mutagenesis at amino acid residue ...

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Site-Directed Mutagenesis of the CP 47 Protein of Photosystem II: Alteration of Conserved Charged Residues in the Domain ³⁶⁴E−⁴⁴⁴R

Cited by 47 publications

References 36 publications

The extrinsic proteins of Photosystem II

The extrinsic proteins of Photosystem II

Intermolecular and Intramolecular Interactions of the 33‐kDa Protein in Photosystem II

The psbo1 Mutant of Arabidopsis Cannot Efficiently Use Calcium in Support of Oxygen Evolution by Photosystem II

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Site-Directed Mutagenesis of the CP 47 Protein of Photosystem II: Alteration of Conserved Charged Residues in the Domain 364E−444R

Cited by 47 publications

References 36 publications

The extrinsic proteins of Photosystem II

The extrinsic proteins of Photosystem II

Intermolecular and Intramolecular Interactions of the 33‐kDa Protein in Photosystem II

The psbo1 Mutant of Arabidopsis Cannot Efficiently Use Calcium in Support of Oxygen Evolution by Photosystem II

Contact Info

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Site-Directed Mutagenesis of the CP 47 Protein of Photosystem II: Alteration of Conserved Charged Residues in the Domain ³⁶⁴E−⁴⁴⁴R