Figure 2. Binding and inhibition of EGFR by Mig6segment 1.
a, Titrations of the wild type EGFR kinase domain and the V924R and I682Q mutants to the 30-residue (residues 334–363) fluorescein-labeled Mig6 peptide. b, Titrations of the wild type EGFR kinase domain to the wild type and three mutant 30-residue fluorescein-labeled peptides. n.d. denotes “not determined” (KD values cannot be determined reliably). c, Inhibition of the activity of the EGFR kinase domain by peptides spanning Mig6segment 1 in the vesicle-based kinase assay. The 60-, 40- and 30-residue peptides contain the entire binding epitope of segment 1, while the 25-residue peptide lacks the N-terminal 3 residues. The mutations were introduced in the 30-residue peptide. See Supplemental Table 1 for the residue boundaries. Fl: fluorescein. d, Cell-based assay showing that Mig6 inhibits full-length EGFR autophosphorylation, whereas mutations in segment 1 abolishes the inhibition.