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Isolation of CP43 and CP47 photosystem II proximal antenna complexes from plants

AuthorsPicorel Castaño, Rafael ; Alfonso Lozano, Miguel ; Seibert, Michael
KeywordsChlorophyll binding protein
Photosystem II
Photosystem II Protein Complex
Photosynthetic Reaction Center Complex Proteins
Plant Proteins
Light-Harvesting Protein Complexes
Anion-exchange chromatography
Issue Date2004
PublisherHumana Press
CitationMethods in Molecular Biology (Clifton, N.J.) 274: 129-135 (2004)
AbstractA single-column method to purify the CP43 and CP47 pigment-protein complexes of photo-system (PS)II from higher plants is presented. To validate the isolation procedure, three different species were used (Spinacea oleracea, Beta vulgaris, and Glycine max), and the procedure worked similarly with all three. Oxygen-evolving core complex obtained from highly enriched PSII membrane fragments were used as the starting material. The core complex is treated with the chaotropic agent LiClO4 and the nonionic detergent n-dodecyl beta-D-maltoside. After dialysis against buffer with no detergent or chaotropic agent, the solubilized material is separated by weak anion-exchange chromatography using a TSK-GEL Toyopearl DEAE 650s column. CP43 complex does not bind to the column and elutes with the first pigmented fractions. When the eluate becomes colorless, the column is subjected to a 0-175 mM LiClO4 linear gradient. The main pigment elution band corresponds to CP47 complex. The last pigmented elution band contains both reaction center-CP47 and reaction center complexes.
Publisher version (URL)http://dx.doi.org/10.1385/1-59259-799-8:129
Identifiersdoi: 10.1385/1-59259-799-8:129
issn: 1064-3745
Appears in Collections:(EEAD) Artículos
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