Studies on the Peroxidase Effect of Cytochrome c. II. Purification of Beef Heart Cytochrome c by Gel Filtration.

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Abstract
A simple and well reproducible procedure for the large scale purification of cytochrome c is described. Final purification was obtained by two consecutive gel filtrations on Sephadex G-75. Cytochrome c was obtained in the native, monomeric form, as indicated by the low percentage of autoxidizable hemoprotein (< 0.5%) and the homogenity in gel filtration. A golden-colored protein impurity, which is always observed during purification of beef heart cytochrome c, was easily and more completely removed by gel filtration than by chromatog-raphy on cation-exchange resins. This fact may explain why the ratios A550 red/A280 ox,A550 red/A550 ox, and A550 red/A535 are higher than those earlier obtained. Some physicochemical properties of the purified preparation are given.