We have compared the dimerization properties and coiled-coil stability of various recombinant fragments of scallop myosin around the head-rod junction. The heavy-chain peptide of the regulatory domain and its various extensions toward the alpha-helical rod region were expressed in Escherichia coli, purified, and reconstituted with the light chains. Rod fragments of the same length but without the light-chain binding domain were also expressed. Electron micrographs show that the regulatory domain complex containing 340 residues of the rod forms dimers with two knobs (two regulatory domains) at one end attached to an approximately 50-nm coiled coil. These parallel dimers are in equilibrium with monomers (Kd = 10.6 microM). By contrast, complexes with shorter rod extensions remain predominantly monomeric. Dimers are present, accounting for ca. 5% of the molecules containing a rod fragment of 87 residues and ca. 30% of those with a 180-residue peptide. These dimers appear to be antiparallel coiled coils, as judged by their length and the knobs observed at the two ends. The rod fragments alone do not dimerize and form a coiled-coil structure unless covalently linked by disulfide bridges. Our results suggest that the N-terminal end of the coiled-coil rod is stabilized by interactions with the regulatory domain, most likely with residues of the regulatory light chain. This labile nature of the coiled coil at the head-rod junction might be a structural prerequisite for regulation of scallop myosin by Ca2+-ions.