The aggregation characteristics of column-purified rabbit skeletal myosin in the presence and absence of C-protein at pH 7.0.

The aggregation properties of column-purified rabbit skeletal myosin at pH 7.0 were investigated as functions of ionic strength, protein concentration, and time. Filaments prepared by dialysis exhibited the same average length and population distribution at 0.10 and 0.15 M KCl at protein concentrations greater than 0.10 mg/ml; similar results were obtained at .0.20 M KCl, although average filament length was approximately 0.5 micrometer shorter. Once formed, these length distributions remained virtually unchanged over an 8-d period. At and below 0.10 mg/ml, average filament length decreased as a function of protein concentration; filaments prepared from an initial concentration of 0.02 mg/ml were half the length of those prepared at 0.2 mg/ml. Filaments prepared by dilution exhibited a sharp increase in average length as the time-course increased up to 40 s, then altered only slightly over a further period of 4 min. Addition of C-protein in a molar ratio of 1-3.3 myosin molecules affected most of these results. Average filament length was affected neither by ionic strength nor by initial protein concentration down to 0.04 mg/ml or over an 8-d period. Filaments formed by dilution in the presence of C-protein exhibited a constant average length and hypersharp length distribution over variable time courses up to 7 min. It is possible that C-protein acts to stabilize the antiparallel intermediate during filamentogenesis, and may also affect subunit addition to this nucleus.