Table 2.

Comparison between measured overall K_D^obs and the calculated macroscopic K_D^kin from kinetic scheme

		KDkin, nM	KDobs, nM
WT	KDbind = 25 ± 4 μM	27 ± 9	26 ± 4
	Keqconf = (11 ± 3) × 10−4
F317L	KDbind = 25 ± 4 μM	42 ± 9	50 ± 10
	Keqconf = (19 ± 2) × 10−4
G250E	KDbind = 25 ± 2 μM	88 ± 26	78 ± 12
	Keqconf = (46 ± 11) × 10−4
Y253F	KDbind = 21 ± 3 μM	53 ± 12	54 ± 4
	Keqconf = (26 ± 5) × 10−4

Comparison of the overall K_D calculated from kinetic data (K_D^kin) with the experimentally measured macroscopic K_D (K_D^obs). Kinetic data are used to calculate individual equilibrium constants for binding (K_D,^bind) and the induced-fit step (K_eq^conf). The overall, kinetic K_D^kin is calculated according to $K_{\mathrm{D}}^{\text{kin}}=K_{\mathrm{D}}^{\text{bind}}\ast K_{\text{eq}}^{\text{conf}}/\left(1+K_{\text{eq}}^{\text{conf}}\right)$ and can be compared to the macroscopic, experimentally determined value, K_D^obs (Fig. 2A) to validate the binding scheme. WT, wild type.