Figure 5

Surface plasmon resonance (SPR) analysis of the Nef-SH3 domain interaction. SPR analyses were performed with the recombinant purified Hck SH3 domains indicated. The SH3 proteins were covalently attached to the biosensor chip, and purified Nef proteins (SF2 and Consensus variants) were serially injected over a range of concentrations. The kinetics and affinities of binding were determined by fitting the sensorgram data (black curves) to a 1:1 Langmuir binding model (red traces) using the BIAevaluation 4.1 software suite. Nef concentrations corresponding to each response curve, along with the calculated binding constants, are presented in Table 1.