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pH-dependent structural change of reduced spinach plastocyanin studied by perturbed angular correlation of γ-rays and dynamic light scattering

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Abstract

In this study the pH-dependent structural changes of reduced spinach plastocyanin were investigated using perturbed angular correlation (PAC) of γ-rays and dynamic light scattering (DLS). PAC data of Ag-substituted plastocyanin indicated that the coordinating ligands are two histidine residues (His37, His87) and a cysteine residue (Cys84) in a planar configuration, whereas the methionine (Met92) found perpendicular to this plane is not a coordinating ligand at neutral pH. Two slightly different conformations with differences in the Cys–metal ion–His angles could be observed with PAC spectroscopy. At pH 5.3 a third coordination geometry appears which can be explained as the absence of the His87 residue and the coordination of Met92 as a ligand. With DLS the aggregation of reduced plastocyanin could be observed below pH 5.3, indicating that not only the metal binding site but also the aggregation properties of the protein change upon pH reduction. Both the structural changes at the metal binding site and the aggregation are shown to be reversible. These results support the hypothesis that the pH of the thylakoid lumen has to remain moderate during steady-state photosynthesis and indicate that low pH induced aggregation of plastocyanin might serve as a regulatory switch for photosynthesis.

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Abbreviations

DLS:

Dynamic light scattering

EFG:

Electric field gradient

Hepes:

N-(2-Hydroxyethyl)piperazine-N′-ethanesulfonic acid

NQI:

Nuclear quadrupole interaction

PAC:

Perturbed angular correlation

Pc:

Plastocyanin

Tris:

Tris(hydroxymethyl)aminomethane

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Acknowledgements

We thank Henrik Vibe Scheller for useful discussions and Marianne Lund Jensen for technical assistance. The project was supported by the Danish Technical Research Council (9901473) and the EU Research Training Network “Transient” (HPRN-CT-1999-00095).

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Authors and Affiliations

  1. Department of Natural Sciences, Royal Veterinary and Agricultural University, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark

    Klára Nárcisz Sas, Lars Hemmingsen & Lars Holm Øgendal

  2. Department of Plant Biology, Royal Veterinary and Agricultural University, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark

    Anna Haldrup

  3. Nano Science Center, University of Copenhagen, Universitetsparken 5, 2100, Købehavn Ø, Denmark

    Eva Danielsen

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  1. Klára Nárcisz Sas
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  2. Anna Haldrup
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  4. Eva Danielsen
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  5. Lars Holm Øgendal
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Correspondence to Klára Nárcisz Sas.

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Sas, K.N., Haldrup, A., Hemmingsen, L. et al. pH-dependent structural change of reduced spinach plastocyanin studied by perturbed angular correlation of γ-rays and dynamic light scattering. J Biol Inorg Chem 11, 409–418 (2006). https://doi.org/10.1007/s00775-006-0085-x

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  • DOI: https://doi.org/10.1007/s00775-006-0085-x

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