Purification and properties of cytochrome c

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Abstract
120 ml. of a soln. of cytochrome c (0.34% Fe) is treated with (NH4)2SO4 until a small quantity remains undissolved. 20 ml. ammonia sp. gr. 0.880 is added to pH 10.0 and let stand overnight at 5[degree]. The precipitate is removed and the filtrate freed of (NH4)2SO4 by dialysis in a cellophane tube for 3 hrs. against tap water and finally against 0.5% KC1. The resulting soln. (250 ml.) is reduced to 50 ml. by suspending in a current of air. It is further dialyzed against KCl and the resulting cytochrome contains 0.43% Fe. Homogeneity of this fraction was established by electrophoresis in a Tiselius apparatus at pH 7.3 and at pH 10.7; by diffusion at pH 7.3; and by chromotog-raphy. Cytochrome prepn. containing 0.34 and 0.43% Fe showed the same catalytic activity/atom of Fe when treated with a kidney prepn. as a source of cytochrome oxidase and succinic dehydrogenase.
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