Regulation of Xanthine Oxidase Activity by Substrates at Active Sites via Cooperative Interactions between Catalytic Subunits: Implication to Drug Pharmacokinetics
Abstract:Three xanthine oxidase substrates (i.e., xanthine, adenine, and 2-amino-4-hydroxypterin) show a “substrate inhibition” pattern (i.e., slower turnover rates at higher substrate concentrations), whereas another two substrates (i.e., xanthopterin and lumazine) show a “substrate activation” pattern (i.e., higher turnover rates at higher substrate concentrations). Binding of a 6-formylpterin at one of the two xanthine oxidase active sites slows down the turnover rate of xanthine at the adjacent active site from 17.0 s-1 to 10.5 s-1, and converts the V-[S] plot from “substrate inhibition” pattern to a classical Michaelis-Menten hyperbolic saturation pattern. In contrast, binding of xanthine at an active site accelerates the turnover rate of 6-formylpterin at the neighboring active site. The experimental results demonstrate that a substrate can regulate the activity of xanthine oxidase via binding at the active sites; or a xanthine oxidase catalytic subunit can simultaneously serve as a regulatory unit. Theoretical simulation based on the velocity equation derived from the extended Michaelis-Menten model shows that the substrate inhibition and the substrate activation behavior in the V-[S] plots could be obtained by introducing cooperative interactions between two catalytic subunits in homodimeric enzymes. The current work confirms that there exist very strong cooperative interactions between the two catalytic subunits of xanthine oxidase.
Keywords: 6-formylpterin; Cooperativity; Michaelis-Menten; binding affinity; buffer solution; catalytic activity; catalytic subunits; concentration; dissociation constant; dissociation rate constants; drug design; enzyme kinetics; enzymes; half life; hetero-substrates; homo-substrates; homo-tetrameric; hyperbolic saturation; hyperbolic saturation region; hyperbolically; isoxanthopterin; kinetic behavior; methylamine dehydrogenase; nitric oxide; nitric oxide formation; oxygen; pH; reactivity; regulation; relative magnitude; substrate activation; substrate inhibition; superoxide; transition point; uric acid; velocity; xanthine; xanthine oxidase
Document Type: Research Article
Publication date: January 1, 2011
- Current Medicinal Chemistry covers all the latest and outstanding developments in medicinal chemistry and rational drug design. Each issue contains a series of timely in-depth reviews written by leaders in the field covering a range of the current topics in medicinal chemistry. Current Medicinal Chemistry is an essential journal for every medicinal chemist who wishes to be kept informed and up-to-date with the latest and most important developments.