The experiments were performed at 37°C on a MicroCal iTC200 titration calorimeter (Malvern Instruments). The reaction cell containing 200 μl of 50 μM proteins was titrated with 19 injections [0.5 μl (first) and 2 μl (all subsequent injections) of 500 μM peptides]. The binding isotherm was fit with Origin 7.0 software package (OriginLab, Northampton, MA) that uses a single set of independent sites to determine the thermodynamic binding constants and stoichiometry.

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