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Isolation of CP43 and CP47 Photosystem II Proximal Antenna Complexes From Plants

  • Rafael Picorel
  • Miguel Alfonso
  • Michael Seibert
Part of the Methods In Molecular Biology™ book series (MIMB, volume 274)

Abstract

A single-column method to purify the CP43 and CP47 pigment-protein complexes of photosystem (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 β-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.

Key Words

Anion-exchange chromatography CP43 CP47 isolation photosystem II pigment-protein complexes 

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Copyright information

© Humana Press Inc., Totowa, NJ 2004

Authors and Affiliations

  • Rafael Picorel
    • 1
  • Miguel Alfonso
    • 1
  • Michael Seibert
    • 2
  1. 1.Estación Experimental de Aula DeiConsejo Superior de Investigaciones CientíficasApdo. ZaragozaSpain
  2. 2.Basic Sciences CenterNational renewable Energy LaboratoryGolden

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