Figure 4. Compound 14 binds to YopH-NT and inhibits full length YopH catalytic activity.
A) Isothermal titration calorimetry data for the binding between YopH-NT and compound 14 (Kd = 310 nM, ΔH = −16.6 Kcal/mol, −TΔS = 7.78 Kcal/mol; n =0.99). B) Western blot analysis for the inhibition by compound 14 of the dephosphorylation of Lck by YopH. Recombinant Lck phosphorylation state on pY394 and total Lck level were detected using selected antibodies, in the absence and presence of compound 14. Full length GST-tagged YopH was pre-incubated with compound 14 for 10 min at room temperature and human recombinant Lck was added and incubated for 20 min.