title: The active site of aromatase cytochrome P-450. Differential effects of cyanide provide evidence for proximity of heme-iron and carbon-19 in the enzyme-substrate complex.
abstract: 19-Norandrostenedione and androstenedione are shown to be metabolized by purified, reconstituted human placental aromatase cytochrome P-450. Kinetic evidence indicates that both steroids share a common catalytic site: 19-norandrostenedione is a competitive inhibitor of androstenedione aromatization, and the Ki value for its inhibition (120 nM) is similar to the Km value for its metabolism (132 nM). The two substrates differ, however, in their sensitivity to inhibition by the heme-iron ligand cyanide; 19-norandrostenedione is approximately 3-fold more sensitive to cyanide inhibition. Spectroscopic studies show that this differential inhibition by cyanide occurs because androstenedione competes with cyanide, whereas 19-norandrostenedione promotes cyanide binding to the heme-iron. It is proposed that these opposite effects on cyanide-iron coordination are due to the proximity of the heme-iron and C-19 of androstenedione in the enzyme-substrate complex, which results in steric exclusion of cyanide from the active site by the C-19 methyl group of androstenedione. Dioxygen is not excluded from binding to the heme-iron during catalysis, presumably because it bonds at an angle, in contrast to the linear bond of iron-cyanide complexes. A model for the active site of aromatase cytochrome P-450 is presented.
mesh_terms: D001141:Aromatase / Q000378:metabolism*; D001665:Binding Sites; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D005260:Female; D006418:Heme / Q000032:analysis; D006801:Humans; D007501:Iron / Q000032:analysis; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology*; D008958:Models, Molecular; D008968:Molecular Conformation; D010920:Placenta / Q000201:enzymology*; D011485:Protein Binding; D013053:Spectrophotometry
chemical_list: D006418:Heme; D003577:Cytochrome P-450 Enzyme System; D007501:Iron; D001141:Aromatase