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Plant and Cell Physiology, 2001, Vol. 42, No. 7 733-741
© 2001 Oxford University Press

Isolation and Characterization of Oxygen-Evolving Thylakoid Membranes and Photosystem II Particles from a Glaucocystophyte, Cyanophora paradoxa

Mari Shibata1, Yasuhiro Kashino, Kazuhiko Satoh and Hiroyuki Koike2

Department of Life Science, Faculty of Science, Himeji Institute of Technology, Harima Science Garden City, Hyogo, 678-1297 Japan

Conditions for preparing oxygen-evolving thylakoid membranes and PSII complexes, and those for observing the PSII activity were investigated in a glaucocystophyte, Cyanophora paradoxa. The active thylakoid membranes were isolated either with a medium containing glycerol or with that containing high concentrations of sucrose, phosphate, and citrate. Active PSII particles were solubilized by octyl-ß-D-glucoside from thylakoid membranes and were separated by sucrose density gradient centrifugation. The thylakoid membranes and PSII particles showed an oxygen-evolving activity only in high-ionic-strength media. The extrinsic 33 kDa protein (PsbO) and the cytochrome c550 (PsbV) were found to be present in the PSII particles as in cyanobacteria or red algae, but no 12 kDa protein (PsbU) was detected. The PsbO protein was classified as a land-plant type by its N-terminal amino acid sequence.

1 Present address: Graduate School of Bioagricultural Science, Nagoya University, Nagoya, 464-8601 Japan.

2 Corresponding author: E-mail, hkoike@sci.himeji-tech.ac.jp; Fax, +81-791-58-0185.


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