Purification and properties of cytochrome f from Brassica komatsuna leaves

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Cytochrome f was extracted from leaves of Brassica komatsuna (Brassica Rapa L. var. pervidis Bailey) in an aqueous solution using methyl ethyl ketone and was purified by the following steps: (i) acetone precipitation, (ii) ammonium sulfate fractionation (0.33-0.7 saturation), (iii) DEAE-cellulose column chromatography, and (iv) Sephadex G-100 column chromatography. Characteristic spectroscopic properties and the midpoint potential of the cyto-chrome were essentially identical with those of the cytochrome f from parsley reported by Bendall et al. Molecular weight of the cytochrome determined by gel filtration was close to 32,000 and it contained one haem per molecule of protein. The ferro-cytochrome was oxidized by potato polyphenol oxidase in the presence of chlorogenic acid. Under light-aerobic conditions, the ferro-cytochrome was rapidly oxidized by the chlorophyll-protein CP743 from Chenopodium album in the presence of menadione. Under light-anaerobic conditions, the oxidized cytochrome was reduced at a consider-able rate.
日本植物生理学会の論文