TABLE 1.

Affinity of CRM1 and variants for YFP-PKI-NES determined using fluorescence anisotropy

Binding constants (K_d) were determined from three independent titrations using the data shown in supplemental Figure S1. The free energy change (ΔΔG) for each variant relative to wild-type CRM1 was calculated from the difference in binding constant.

CRM1 protein	Kd	ΔΔG
	μm	kcal/mol
Wild-type	15.6 ± 2.3
1–1062	1.7 ± 0.1	1.3 ± 0.2
VLV > AAA	0.20 ± 0.03	2.6 ± 0.2
1–1027	0.30 ± 0.05	2.3 ± 0.2
1–1062 + VLV > AAA	0.025 ± 0.002	3.8 ± 0.2