CRYSTALLINE CYTOCHROME C

IV. CRYSTALLIZATION OF BEEF KIDNEY CYTOCHROME C

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kidney tissues contain an impure protein which prevents sufficient purification of cytochrome c for crystallization by any of the methods reported for muscle tissues. This impurity possessing a similar affinity to the resin to that of oxidized cytochrome c was found to be removed by chromatography of the cytochrome solution in its reduced form permitting easy crystallization of the pigment. On the basis of this finding, two methods, i.e., Methods A and B, were described for purification and crystallization of kidney cytochrome c. These two methods are the mo-difications of Method III (using direct adsorption) reported in the previous papers. In the latter method the yield of crystals was about 40mg. from 6kg. of kidneys (3kg. of cortex), while the former method gave a smaller yield because of modification of the pigment caused during the TCA precipitation. Crystals usually appeared as rosettes similar to those of beef heart cytochrome c and sometimes as hexagonal plates.