FIGURE 3.

Role of Tyr⁵¹⁰ in binding of Dbl to Grb2. COS7 cells were co-transfected with indicated constructs, and association was evaluated as described under “Materials and Methods.” A, association of Grb2 with Dbl or Dbl (Y510F). B, association of Grb2 with Dbl is dependent on tyrosine phosphorylation. Experiments were done as described in A, in the presence or absence of the tyrosine phosphate inhibitor sodium orthovanadate (1 mm). Top panels, presence of Grb2 and Dbl in anti-Dbl immunoprecipitates. Bottom panels, protein expression in cell lysates. Data are representative of three independent experiments. IB, immunoblot.