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Fluorescence Polarization Screening Assays for Small Molecule Allosteric Modulators of ABL Kinase Function

Fig 11

Compound 142 fails to activate downregulated HCK in vitro.

Top panels: Recombinant, downregulated wild type ABL core and near full-length HCK proteins were assayed in the presence of compound 142 (1 μM) or with DMSO alone as control (Con) using a kinetic kinase assay (see Materials and Methods). Data are plotted as pmol ADP produced per ng kinase as a function of time. Bottom: Amino acid sequence alignment of the ABL and HCK SH3 domains and SH2-kinase linkers. SH3 and linker residues predicted to contribute to compound 142 binding are highlighted in red (see Fig 10). The type of interaction is indicated as side chain (s), main chain (m), or aromatic (a).

Fig 11