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. 2015 Mar 30;290(20):12879–12898. doi: 10.1074/jbc.M114.634493

FIGURE 4.

FIGURE 4.

UC-773587 and UC-857993 inhibit GEF catalytic activity. A, chemical structures of the two hit compounds, UC-773587 and UC-857993. UC-773587 and UC-857993 dose-dependently inhibit (B and D) 50 nm His6-SOS1cat (black circle)-mediated BODIPY-FL-GDP for GTP nucleotide exchange on 2 μm His6-H-Ras (residues 1–166) (gray circle) or (C and E) 50 nm His6-SOS1cat (black circle)-mediated BODIPY-TR-GTP loading on 2 μm His6-H-Ras (residues 1–166) (gray circle) in a 96-well format at the indicated compound concentrations. F, UC-773587 (3 μm) and UC-857993 (20 μm) additively inhibit 50 nm His6-SOS1cat (black circle)-mediated BODIPY-FL-GDP for GTP nucleotide exchange on 2 μm His6-H-Ras (residues 1–166) (gray circle) in a 96-well format. Data in B–F are expressed as percent change in fluorescence units normalized to the initial time point and were performed in triplicate. Data in B–E represent the mean of three independent experiments.