Evidence from electron microscope studies on actin paracrystals concerning the origin of the cross-striation in the thin filaments of vertebrate skeletal muscle

Abstract

Abstract When purified F-actin is precipitated by Mg2+, it forms paracrystals consisting of regularly packed filaments that retain the double-helical structure characteristic of actin polymers. In negatively stained preparations the only axial periodicity observed is that of the actin polymers. In sections, the polymer structure is not resolved and the paracrystals do not appear cross-striated. The paracrystals formed from unpurified actin preparations likewise contain regularly packed filaments in which the F-actin structure is observed but, in addition, the assembly of filaments is crossed at fairly regular intervals (approximately 37 nm) by bands of unstained amorphous material. Sections of these paracrystals show a corresponding cross-striation (mean spacing 38 nm). Other proteins known to be present in unpurified preparations of actin were prepared separately and added in solution to purified F-actin; paracrystals were formed on Mg2+ precipitation. The cross-striation was reproduced when a mixture of tropomyosin and troponin had been added to the actin. The addition of purified tropomyosin alone resulted in paracrystals that lacked the cross-striation. Comparison of the cross-striation in the paracrystals with that in the thin filament assembly of the myofibril (as seen in sections and in negatively stained I-segments) supports the conclusion that filaments resembling the natural ones have been synthesized from actin, tropomyosin and the troponin complex. It follows that the cross-striation in the thin myo­filaments can be attributed to the location of part or all of the troponin complex at sites spaced at regular intervals along the filaments.