The side-chain cleavage of cholesterol sulfate--I. The effect of adrenodoxin on the binding of cholesterol sulfate to cytochrome P-450scc.
Difference spectroscopy was used to measure the binding of cholesterol sulfate (CS) to cytochrome P-450scc. The uncomplexed cytochrome and the complex of the cytochrome with adrenodoxin (ADX) were both titrated with CS in order to test whether ADX increased the affinity of the cytochrome for the sterol sulfate. The addition of ADX to the cytochrome had different effects on the binding of the sterol sulfate depending on several factors including: (1) The method of preparation of the cytochrome P-450scc, (2) The concentration of cytochrome P-450scc, (3) The method by which CS was suspended in aqueous solution, and (4) Whether or not the solutions of cytochrome contained non-ionic detergents. The results of this study suggest that the method of isolation of cytochrome P-450scc, and non-ionic detergents, greatly modulate the apparent affinity of cytochrome P-450scc for CS. In the absence of detergents the addition of adrenodoxin to dilute solutions of cytochrome P-450scc appears to enhance only slightly (1- to 2-fold) the affinity of the cytochrome for the sterol sulfate.
D000302:Adrenal Cortex / Q000032:analysis; D000325:Adrenodoxin / Q000494:pharmacology*; D001665:Binding Sites; D002784:Cholesterol / Q000378:metabolism; D002788:Cholesterol Esters / Q000378:metabolism*; D019826:Cholic Acid; D002793:Cholic Acids / Q000494:pharmacology; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D003902:Detergents / Q000494:pharmacology; D008928:Mitochondria / Q000032:analysis; D011136:Polysorbates / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D012996:Solutions; D013053:Spectrophotometry
D002788:Cholesterol Esters; D002793:Cholic Acids; D003902:Detergents; D011136:Polysorbates; D012996:Solutions; D000325:Adrenodoxin; D003577:Cytochrome P-450 Enzyme System; D002784:Cholesterol; D019826:Cholic Acid; C007045:cholesteryl sulfate