... solution.In order to determine the affinity for Ca21 and the cooperativity of Ca21-binding to p26olf, we next analyzed the stoichiometry of Ca21-binding to p26olf by flow dialysisin the Tris buffer. ... sequentially, and has a total of four Ca21-bindingsites (known as EF-hands). In this study, to elucidate the mechanism of Ca21-binding to each EF-hand (named EF-A,-B, -C and -D from the N-terminus ... suggested that the orderof Ca21-binding to p26olf is EF-B, EF-A, EF-C and EF-D.In these EF-hands, EF-B and -D are the p26olf-specificEF-hands and probably show high affinity for Ca21-binding,while...