Fig. 1. Phosphorylation of T474-FAK suppresses FAK autophosphorylation. In vitro kinase assays were performed using FLAG-tagged WT or mutant proteins of which indicated threonine (T) residue were mutated to glutamate (E), (T→E), and [32P]-ATP. FLAG-WT FAK (FLAG-FAK) or six FLAG-tagged mutants (T219, T227, T284, T455, T474, or T979→E) were overexpressed in HEK293T cells and collected by immunoprecipitation, and 32P-labeled FAK was detected by autoradiography. Coomassie blue staining shows the amounts of proteins in each reaction mixture.
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