Slip of rabbit striated muscle in rigor or AMPPNP

Summary

Single glycerol-extracted rabbit psoas muscle fibres have been slowly extended either in rigor or in the unhydrolysable ATP analogue AMPPNP, and their sarcomere length, sarcomere structure and tension measured. The length of regularly arrayed sarcomeres, measured by optical diffraction, increased continuously as the muscle was stretched; the maximum sarcomere extension seen was approximately 6%. In the electron microscope sarcomeres from extended muscle fixed in rigor or AMPPNP remained regular in their internal structure, without rupture or obvious lengthening around the Z line. During steady extension at 0.024% per min the tension in the muscle fibre rose until it reached a limiting value [T _m ] when the sarcomeres had stretched by 0.8–1.6% and then remained constant with continued extension, while the sarcomeres continued to stretch. Provided that a novel form of preparation of the glycerol-extracted fibres was employed,T _m in rigor was a large fraction of the tension expected from an active isometric muscle fibre. In the presence of AMPPNPT _m was reduced by a factor of 2 to 3. Step extension by 0.08% at 5-min intervals gave the same pattern of mechanical response with similar values ofT _m . The isometric tension decay in the interval between the steps was very rapid at first and slowed continuously until the next step. The average speed of tension fall between 30 and 300 s after stretch was measured at each step and plotted relative to the tension in the muscle. The relationship approached linearity, although with a significant upward curvature at high tension. The proportionality constant of the rate of tension fall to tension was 4.5×10⁻⁴ s⁻¹ in rigor and 9×10⁻⁴ s⁻¹ in AMPPNP. These values are less than the apparent dissociation rate constants for acto-subfragment-1 or acto-heavy meromyosin under similar conditions (Marston, 1982). These results indicate that interfilament slip does occur in rabbit skeletal muscle both in rigor and in AMPPNP, but that it is much slower that that predicted from the behaviour of the isolated proteins, as if the myosin heads interacted so as to obstruct their detachment.