Effect of ATP depletion on the isolated thick filament of limulus striated muscle.

With the dynamic light scattering method, we have shown that calcium ions increase the high-frequency internal motions of isolated thick filaments of Limulus striated muscle in the presence of ATP (Kubota, K., B. Chu, Shih-fang Fan, M.M. Dewey, P. Brink, and D. Colflesh. J. Mol. Biol. 1983. 166:329 and Fan, S.-f., M.M. Dewey, D. Colflesh, B. Gaylinn, R. Greguski, and B. Chu. Biophys. J. 1985. 47:809). If ATP is removed from the suspending medium, an increase of high-frequency internal motions also has been observed with characteristics different from those of filaments suspended in a medium containing ATP and calcium ions. These internal motions appear whether calcium ions are present or not and are suppressed by trifluoperazine (TFP). The motions differ from the calcium ion-induced motions in that (a) an energy supply is not required; (b) they are insensitive to heat treatment (42 degrees C, 10 min) and (c) they are also insensitive to phenylmethylsulfonyl fluoride which blocks the motions in the presence of ATP and calcium ions (Fan, Shih-fang, M.M. Dewey, D. Colflesh, B. Gaylinn, R. Greguski, and B. Chu. 1985. Biochim. Biophys. Acta. 827:101). Electron micrographs of negatively stained thick filaments in an ATP-free medium show that the majority of crossbridges extend out from the backbone of the filament and optical diffraction patterns from these filaments lack layer lines arising from the crossbridges. The flexibility of the thick filaments suspended in ATP-free media increases.