Impact of enzyme concentration and residence time on apparent activity recovery in jump dilution analysis.

摘要: Jump dilution analysis is commonly used to evaluate the reversibility of inhibition and quantify residence time inhibitor-enzyme complex. During hit lead characterization, one sometimes observes apparently linear progress curves after jump that display activity recoveries are intermediate between those expected for fully reversible irreversible inhibition. Computer simulations indicate seemingly can result when dealing with tight-binding inhibitors if substoichiometric concentrations inhibitor preincubated enzyme. In this situation, recovered comparable instantaneously inhibitors. addition, demonstrate values recovery may be observed compounds modestly slow dissociation rates (i.e., times >0min but ≤20min) attending curvature data not accounted for. The observation can, thus, serve as an indication either modest or a contaminating inactivator within sample, in case prompting greater scrutiny test compound.