Affinity-purified melittin antibody recognizes the calmodulin-binding domain on calmodulin target proteins.

Melittin is a 26-amino acid amphipathic peptide which binds to calmodulin in a calcium-dependent manner. The utility of melittin as a peptide replica of the calmodulin-binding region of calmodulin acceptor proteins (CaMBPs) was investigated. Antibody against melittin was raised and purified by antigen affinity chromatography. Interaction of the antibody with CaMBPs was initially suggested by the ability of anti-melittin-Sepharose, but not nonimmune IgG-Sepharose, to bind calmodulin-dependent cyclic AMP phosphodiesterase. Direct interaction of melittin antibody with the calmodulin-binding domain of acceptor proteins was demonstrated by quantitative inhibition of calmodulin binding to the purified CaMBPs, myosin light chain kinase, and eel electric organ CaMBP55. These results indicate that melittin antibody identifies regions of structural similarity between calmodulin acceptor proteins, and this region includes a common calmodulin-binding domain.