Journal Article

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

Mari Shibata, Yasuhiro Kashino, Kazuhiko Satoh and Hiroyuki Koike

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 42, issue 7, pages 733-741
Published in print July 2001 | ISSN: 0032-0781
Published online July 2001 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pce092
Isolation and Characterization of Oxygen-Evolving Thylakoid
Membranes and Photosystem II Particles from a Glaucocystophyte,
Cyanophora
paradoxa

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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-β-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.

Keywords: Key words: Cyanophora paradoxa — Glaucocystophyte — Oxygen evolution — Photosystem II — PsbO — Thylakoid membranes.; Abbreviations: DCIP, 2,6-dichlorophenolindophenol; DPC, 1,5-diphenylcarbazide; DM, n-dodecyl-β-d-maltoside; EGTA, O,O′-bis(2-aminoethyl) ethyleneglycol-N,N,N′,N′-tetraacetic acid; MV, methyl viologen; OG, n-octyl-β-d-glucoside; p-BQ, p-benzoquinone; SPC, 0.5 M sucrose, 0.5 M phosphate and 0.3 M citrate (pH 6.9).

Journal Article.  6188 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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