| Assay Method Information | |
| | Measurement of Inhibition Constants |
| Description: | Inhibition constants (Ki) were calculated by determining the kcat and Km (DDCoA) values at several fixed inhibitor concentrations. The inhibition data were analyzed using the standard equation for uncompetitive inhibition. For compounds with Ki values in the low nanomolar range, initial velocities were determined at a fixed substrate concentration and the data were fit to the equation: vi/v0 = (Km + S)/(Km + S[1 + I/ Ki]). Where vi and v0 are the initial velocities in the presence and absence of inhibitor. The substrate concentration was fixed at 15 uM, and the inhibitor concentration was varied from 3.5 to 1200 nM. Data fitting was performed using Grafit 4.0 (Erithacus Software Ltd.). |
| Affinity data for this assay | |
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