FIGURE 9.

SPR analysis of Nef·Hck32 interaction. SPR analyses were conducted with recombinant purified wild-type Hck32 (top) and the Hck32 SH3 E93A mutant (bottom). Recombinant purified Nef was covalently attached to the SPR chip surface, and Hck32 proteins were injected over the range of concentrations shown. The kinetics and affinities of binding were determined by fitting the sensorgram data (black traces) to a 1:1 binding model (red curves) using the Biacore T100 evaluation (2.0.4) software. The resulting binding constants are summarized below the graphs.