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Calmodulin (CaM) is a small (148 residues), acidic, and highly conserved calcium binding protein found in all eukaryotic cells. In order to understand the role of calmodulin in deciphering the calcium signal, it is necessary to analyze the ion binding mechanism of the protein. The apoCaM (Ca2+-free CaM) is the background state and probably plays an important role in the cell. As is known, it can interact, although much more weakly, with some target proteins such as neuromodulin and brush border myosin I. It is of paramount importance to describe this state from a structural and thermodynamic point of view.