Thermodynamic Analysis of Calmodulin-CDZ Interaction

ITC experiments were performed using a VP-ITC calorimeter (Malvern Panalytical, Orsay, France). The ITC experiments were performed in buffer F at 25°C (unless otherwise stated). For a typical titration, the solution of analyte (8 μM of holo-CaM) was loaded in the reaction cell. The titrant (200 μM of CDZ) was loaded into the syringe before injection of 5 to 15 μl of the titrant into the reaction cell at intervals of 600 s. Heats of dilutions were measured by injecting the titrant into the buffer and were subtracted from the heat of reaction. The titration profiles were analyzed using the Origin7 software (OriginLab, Northampton, MA, USA) to determine the thermodynamic parameters, as previously described [35 (link)].