Structural investigation of PsbO from plant and cyanobacterial photosystem II |
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Authors: | Slowik Daria Rossmann Maxim Konarev Petr V Irrgang Klaus-D Saenger Wolfram |
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Affiliation: | 1 Max Volmer Laboratory of Biophysical Chemistry, Technische Universität Berlin, Strasse des 17, Juni 135, 10623 Berlin, Germany2 Institute of Chemistry and Biochemistry/Crystallography, Freie Universität Berlin, Takustr. 6, 14195 Berlin, Germany3 European Molecular Biology Laboratory, Hamburg Outstation Building 25A, Notkestr. 85, 22603 Hamburg, Germany4 Department of Life Sciences and Technology, Beuth Hochschule für Technik Berlin, Seestr. 64, 13347 Berlin, Germany |
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Abstract: | The manganese-stabilizing protein PsbO is associated with the luminal side of thylakoids close to the redox-active Mn4Ca cluster at the catalytically active site of photosystem II (PSII). PsbO is believed to increase the efficiency of oxygen evolution and to stabilize the Mn4Ca cluster against photoinhibition. Using small-angle X-ray scattering, we investigated the low-resolution structure of wild-type spinach PsbO and that of chimeric spinach PsbO fused with maltose-binding protein. Small-angle X-ray scattering data revealed that both proteins are monomeric in solution, and that plant and cyanobacterial PsbO have similar structures. We show a highly efficient expression system that allows recombinant production of the active wild type and the chimeric PsbO from spinach and cyanobacteria, with yields compatible with biophysical and structural studies. The binding of spinach PsbO fused with maltose-binding protein to PSII depleted of extrinsic subunits (PSII-ΔpsbO,P,Q) was confirmed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. The reconstituted PSII was shown to have similar oxygen evolution rates as obtained with wild-type spinach PsbO. |
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Keywords: | PSII, photosystem II MBP, maltose-binding protein MALDI-TOF-MS, matrix-assisted laser desorption ionization time-of-flight mass spectrometry SAXS, small-angle X-ray scattering wt, wild-type spinach MM, molecular mass TRX, thioredoxin Mes, 4-morpholineethanesulfonic acid |
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