Characterization of a calmodulin antiserum by its reactions with fragments of the calmodulin molecule

A high affinity antibody, specific to the calcium-free form of calmodulin, which had previously been developed using N-acetyl-muramyl-L-alanyl-D-isoglutamine-calmodulin conjugate as an immunogen, was tested for cross-reactivity with tryptic fragments of calmodulin (CaM1-77, CaM1-90, CaM78-149, and CaM106-149) as well as with synthetic peptides corresponding to the 1st, 2nd, and 3rd calcium binding loop of calmodulin. The results showed that the antigenic determinant involves a special conformation of amino acid residues 90-106 in the 3rd calcium-binding domain.