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Figure 1


Fig. 1. Analysis of GST-Grp1-PH binding by time-resolved FRET. (A) FRET saturation-binding isotherms. Protein (20 nM) was incubated with increasing concentrations of biotinylated PtdIns(3,4,5)P3. (B) IC50s of binding were measured by incubating FRET complexes containing 80 nM GST-Grp1-PH with increasing concentrations of the indicated unlabeled ligands. Data are presented as a mean of three independent experiments ± s.e.m. Values measured from A and B were used to calculate Ki values (see Materials and Methods) shown in C.