title: Direct electrochemistry of human, bovine and porcine cytochrome P450c17.
abstract: The direct electrochemistry of human, bovine and porcine cytochrome P450c17 (CYP17) has been examined on an edge-oriented pyrolytic graphite electrode. The recombinant protein was immobilized on an electrode modified with a surfactant to simulate the environment of a biological membrane, and hence physiological electron-transfer conditions. The P450 enzymes all retained 'electron-transfer' activity while immobilized at the electrode surface as assessed by the presence of catalytic signals under aerobic conditions. The redox potentials for porcine P450c17 were more positive (anodic) than both the human and bovine forms, perhaps reflecting the differences in substrate specificity for these species. In addition, these enzymes were all influenced by pH, consistent with a single proton associated with the single electron-transfer event. Ionic strength of the buffer medium also shifted the redox potentials towards positive, suggesting that electrostatic forces contribute to the protein environment required for the electron-transfer process. The effect of substrate on the redox potential for each P450c17 was measured in the presence of pregnenolone, progesterone, 17alpha-hydroxypregnenolone and 17alpha-hydroxyprogesterone. However, no influence on the redox parameters was observed.
authors.affiliation: School of Chemistry, Monash University, Victoria, 3800, Australia.
authors.forename: D L
authors.initials: DL
authors.lastname: Johnson
authors.forename: A J
authors.initials: AJ
authors.lastname: Conley
authors.forename: L L
authors.initials: LL
authors.lastname: Martin
mesh_terms: D000818:Animals; D002417:Cattle; D004563:Electrochemistry; D004566:Electrodes; D004800:Enzymes, Immobilized / Q000235:genetics / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D013254:Steroid 17-alpha-Hydroxylase / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D013552:Swine
chemical_list: D004800:Enzymes, Immobilized; D013254:Steroid 17-alpha-Hydroxylase