Heteroactivator effects on the coupling and spin state equilibrium of CYP2C9.
The cytochromes P450 are capable of oxidizing a variety of xenobiotics. Binding of a small molecule heteroactivator to a P450 can alter the coupling of substrate oxidation during P450 catalysis, but the degree to which coupling or shunting via one of the three catalytic cycle branch points is linked to the heteroactivator-modified position of bound substrate is unknown. Using reconstituted CYP2C9, stoichiometric measurements were gathered with three substrates and two classes of heteroactivators to further understand the mechanisms involved in heteroactivation. Heteroactivation of P450 metabolism appeared to involve, but not require, changes in coupling and that increased uncoupling to a specific byproduct like H(2)O(2) does not necessarily correlate to the degree of coupling. In addition, spectroscopy demonstrated that every heteroactivator tested influenced the spin equilibrium of the heme iron even in the presence of saturating substrate suggesting that both substrate proximity and the ability to desolvate the heme can be involved in heteroactivation.
University of Minnesota, Department of Experimental and Clinical Pharmacology, USA.
Charles W
CW
Locuson
Peter M
PM
Gannett
Timothy S
TS
Tracy
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000737:chemistry*; D001189:Aryl Hydrocarbon Hydroxylases / Q000032:analysis / Q000737:chemistry*; D001665:Binding Sites; D003198:Computer Simulation; D065729:Cytochrome P-450 CYP2C9; D004789:Enzyme Activation; D006861:Hydrogen Peroxide / Q000737:chemistry*; D008956:Models, Chemical; D008958:Models, Molecular; D011485:Protein Binding; D013113:Spin Labels; D013379:Substrate Specificity
D000894:Anti-Inflammatory Agents, Non-Steroidal; D013113:Spin Labels; D006861:Hydrogen Peroxide; C450260:CYP2C9 protein, human; D065729:Cytochrome P-450 CYP2C9; D001189:Aryl Hydrocarbon Hydroxylases