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Quantitative analysis of NES-CRM1 affinity in vitro using the RanGAP hydrolysis assay. Inhibitory effect of LMB (1 μM; ▵), PKF050-638 (330 μM; ▴), PKF050-637 (330 μM; ×), and no drug (♦) on CRM1-NS2 (A) or CRM1-PKI (B) binding as measured from CRM1-dependent protection of Rna1p-stimulated GTP hydrolysis on Ran, as a function of increasing concentrations of CRM1. In all series, NS2 or PKI peptides are present at 40 and 200 μM, respectively.
