Bovine adrenal steroid hydroxylase system III. Reconstitution of adrenal iron-sulfur protein

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1. 1. The apo adrenal ferrodoxin has been prepared by treatment of native adrenal ferrodoxin with trichloroacetic acid. 2. 2. The recombination reaction of the apoprotein with Fe2− and sulfide in the presence of 2-mercaptoethanol was greatly enhanced by the presence of a high concentration of urea, up to 75% recovery. 3. 3. Crystals of the reconstituted protein were prepared followed by chromatography on DEAE-cellulose. 4. 4. The reconstituted adrenal ferredoxin was homogeneous on ultracentrifugation and on disc electrophoresis and was indistinguishable from the native material in terms of its catalytic functions, absorption spectrum, molecular weight and the iron content. 5. 5. A method has been described for measuring the labile sulfide in adrenal ferredoxin. The sulfide content was the same in both preparations.

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