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. 2008 May 30;283(22):15491–15501. doi: 10.1074/jbc.M800002200

FIGURE 2.

FIGURE 2.

The MbSrc1 SH2 and SH3 domains are functional. A, immobilized GST-SH2, GST-SH3, or GST were incubated with Src-transfected SYF cell lysate for 1 h at 4 °C. Competitor peptides were added to the incubations as indicated. The GST proteins were collected by centrifugation, washed with lysis buffer, and eluted with SDS-PAGE sample buffer. Bound proteins were visualized by anti-Tyr(P) Western blotting. The left two lanes show 25 μg of Src-transfected and untransfected SYF lysate, respectively, loaded directly onto the gel. B, binding of the MbSrc1 SH2 domain to pYEEI peptide measured by ITC. A representative titration is shown using 60 μm SH2 domain and 5-μl injections of 1.8 mm pYEEI peptide. The dissociation constant determined from this titration was 140 nm.