Calcium-dependent structural changes in scallop heavy meromyosin.

The mechanism of calcium regulation of scallop myosin is not understood, although it is known that both myosin heads are required. We have explored possible interactions between the heads of heavy meromyosin (HMM) in the presence and absence of calcium and nucleotides by sedimentation and electron microscope studies. The ATPase activity of the HMM preparation was activated over tenfold by calcium, indicating that the preparation contained mostly regulated molecules. In the presence of ADP or ATP analogs, calcium increased the asymmetry of the HMM molecule as judged by its slower sedimentation velocity compared with that in EGTA. In the absence of nucleotide the asymmetry was high even in EGTA. The shift in sedimentation occurred with a sharp midpoint at a calcium level of about 0.5 microM. Sedimentation of subfragment 1 was not dependent on calcium or on nucleotides. Modeling accounted for the observed sedimentation behavior by assuming that both HMM heads bent toward the tail in the absence of calcium, while in its presence the heads had random positions. The sedimentation pattern showed a single peak at all calcium concentrations, indicating equilibration between the two forms with a t(1/2) less than 70 seconds. Electron micrographs of crosslinked, rotary shadowed specimens indicated that 81 % of HMM molecules in the presence of nucleotide had both heads pointing back towards the tail in the absence of calcium, as compared with 41 % in its presence. This is consistent with the sedimentation data. We conclude that in the "off" state, scallop myosin heads interact with each other, forming a rigid structure with low ATPase activity. When molecules are switched "on" by binding of calcium, communication between the heads is lost, allowing them to flex randomly about the junction with the tail; this could facilitate their interaction with actin in contracting muscle.