Organization of the oxygen-evolution enzyme complex studied by butanol/water phase partitioning of spinach photosystem II particles.

The oxygen-evolving photosystem II particles prepared from spinach chloroplasts with brief sonication and Triton X-100 treatment were subjected to butanol/water phase partitioning. Three peripheral proteins of photosystem II having relative molecular masses of 33,000, 24,000, and 18,000 daltons and a part of the manganese atoms associated with photosystem II were partitioned into the aqueous phase, depending on the concentration of salt which was included in the suspension of the photosystem II particles. Quantitative analysis of the phase partitioning of the photosystem II particles under the various ionic conditions at pH 6.5 suggested the following: (a) two of the four atoms of manganese associated with photosystem II are located at a relatively hydrophilic environment and easily extracted from the membrane; (b) one of these "hydrophilic manganese atoms" is structurally in close proximity to the protein of the relative molecular mass of 33,000 daltons and stabilized by the protein specifically; (c) the protein of the relative molecular mass of 24,000 daltons as well as that of 33,000 daltons is involved in the stabilization of the other "hydrophilic manganese" in the membrane; (d) each of the three proteins has an independent binding site on the membrane and organizes a specific catalytic domain where oxidation of water is carried out efficiently in collaboration with the reaction center of photosystem II.