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High-Throughput Screening of Enzyme Inhibitors: Simultaneous Determination of Tight-Binding Inhibition Constants and Enzyme Concentration

✍ Scribed by Petr Kuzmič; Kyle C Elrod; Lynne M Cregar; Steve Sideris; Roopa Rai; James W Janc

Book ID
102563940
Publisher
Elsevier Science
Year
2000
Tongue
English
Weight
74 KB
Volume
286
Category
Article
ISSN
0003-2697

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✦ Synopsis

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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