Oxygen-Evolving Photosystem II Preparation from Wild Type and Photosystem II Mutants of Synechocystis Sp. PCC 6803

Diana L. Kirilovsky, Alain G.P. Boussac, Frans J.E. van Mieghem, Jean Marc R.C. Ducruet, Pierre R. Sétif, A. William Rutherford, Jiujiang Yu, Wim F.J. Vermaas

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Abstract

We present here a simple and rapid method which allows relatively large quantities of oxygen-evolving photosystem II-(PS-II-) enriched particles to be obtained from wild-type and mutants of the cyanobacterium Synechocystis 6803. This method is based on that of Burnap et al. [Burnap, R., Koike, H., Sotiropoulou, G., Sherman, L.A., & Inoue, Y. (1989) Photosynth. Res. 22, 123-130] but is modified so that the whole preparation, from cells to PS-II particles, is achieved in 10 h and involves only one purification step. The purified preparation exhibits a 5-6-fold increase of O2-evolution activity on a chlorophyll basis over the thylakoids. The ratio of PS-I to PS-II is about 0.14:1 in the preparation. The secondary quinone electron acceptor, QB, is present in this preparation as demonstrated by thermoluminescence studies. These PS-II particles are well-suited to spectroscopic studies as demonstrated by the range of EPR signals arising from components of PS-II that are easily detectable. Among the EPR signals presented are those from a formal S3-state, attributed to an oxidized amino acid interacting magnetically with the Mn complex in Ca2+-deficient PS-II particles, and from S2 modified by the replacement of Ca2+ by Sr2+. Neither of these signals has been previously reported in cyanobacteria. Their detection under these conditions indicates a similar lesion caused by Ca2+ depletion in both plants and cyanobacteria. The protocol has also been applied to mutants which have site-specific changes in PS-II. Data are presented on mutants having changes on the electron donor (Y160F) and electron acceptor (G215W) side of the D2 polypeptide.

Original languageEnglish (US)
Pages (from-to)2099-2107
Number of pages9
JournalBiochemistry
Volume31
Issue number7
DOIs
StatePublished - Feb 1 1992
Externally publishedYes

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ASJC Scopus subject areas

  • Biochemistry

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Kirilovsky, D. L., Boussac, A. G. P., van Mieghem, F. J. E., Ducruet, J. M. R. C., Sétif, P. R., Rutherford, A. W., Yu, J., & Vermaas, W. F. J. (1992). Oxygen-Evolving Photosystem II Preparation from Wild Type and Photosystem II Mutants of Synechocystis Sp. PCC 6803. Biochemistry, 31(7), 2099-2107. https://doi.org/10.1021/bi00122a030