Figure 2.

Tyrosine phosphorylated PKL binds to Nck and Src. (A) Cultured SYF vector control and Src replete MEFs were lysed followed by anti-Nck or anti-phosphotyrosine 4G10 IP and anti-PKL or 4G10 immunoblotting (IB) revealing PKL is tyrosine phosphorylated and coimmunoprecipitates with Nck. (B) PKL binds to the Nck and Src SH2 domains in an adhesion-dependent manner. SYF or SYF + Src cells were maintained in suspension or replated on fibronectin for 20 min followed by anti-Nck immunoprecipitation (IP), incubation with GST, GST-NckSH2, or GST-SrcSH2 followed by PKL IB. (C) PKL can also bind constitutively to the SH3 domains of Nck and Src.