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  • Title: High-throughput screening of enzyme inhibitors: simultaneous determination of tight-binding inhibition constants and enzyme concentration.
    Author: Kuzmic P, Elrod KC, Cregar LM, Sideris S, Rai R, Janc JW.
    Journal: Anal Biochem; 2000 Nov 01; 286(1):45-50. PubMed ID: 11038272.
    Abstract:
    Active site titration by a reversible tight-binding inhibitor normally depends on prior knowledge of the inhibition constant. Conversely, the determination of tight-binding inhibition constants normally requires prior knowledge of the active enzyme concentration. Often, neither of these quantities is known with sufficient accuracy. This paper describes experimental conditions under which both the enzyme active site concentration and the tight-binding inhibition constant can be determined simultaneously from a single dose-response curve. Representative experimental data are shown for the inhibition of human kallikrein.
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