FIGURE 1.
Analyses of the effect of glycerol on the Kd of the cpSRP43-A3CT complex using ITC. A, binding isotherms measured by ITC at 20 °C for the titration of ITC Buffer into ITC Buffer. No significant heat change associated with the injections was observed. ITC Buffer: 20 mm Hepes/NaOH pH 7.5, 200 mm NaCl, 2 mm MgCl2, 1 mm EDTA, 5% (w/v) glycerol, 0.25 mm tris(2-carboxyethyl)phosphine). B, determination of binding affinities for the cpSRP43-A3CT interaction by ITC in the absence of glycerol. The titration was performed at 20 °C in ITC Buffer without glycerol. Analysis of the titration isotherms resulted in a Kd of 13.2 μm and a stoichiometry of 1:1. Data analysis was performed using the Origin 7.0 software.