Myosin heads contact with thin filaments in compressed relaxed skinned fibres of frog skeletal muscle.
When skinned skeletal muscle fibres with rest sarcomere length (L = 2.5 microns) are compressed by the addition of various concentrations ([PVP]) of polyvinylpyrrolidine, the relation between the 1,0 spacing (d) of thick filament lattice and [PVP] has been known to break at d of around 35 nm, resulting in a steeper slope of the relationship at d greater than 35 nm. To clarify the cause of this, X-ray diffraction and crosslinking experiments were carried out. The d versus [PVP] relationship of stretched fibres (L = 3.5 microns) breaks at a d of around 29 nm. The difference between these characteristic d values, 35-29 = 6 nm, is close to the diameter of thin filaments (8 nm). The crosslinking efficiency of formaldehyde between myosin heads and thin filament surface, measured by radial stiffness increase, was found to begin to markedly increase when the relaxed fibre with rest L was compressed to a d of nearly 35 nm. In addition to these results, the d versus [PVP] relationship obtained in rigor and in high [Mg2+] (30 mM) relaxing solutions, and the crosslinking efficiency seen in high [Mg2+] solutions supported our previous hypothesis that in normal relaxing solution (containing 1 mM Mg2+) the probability of myosin heads coming into contact with the thin filament surface abruptly increases at d near 35 nm in fibres with rest L.
Umazume
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Y
Department of Physiology, Jikei University School of Medicine, Tokyo, Japan.
Higuchi
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H
Takemori
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S
D000255:Adenosine Triphosphate / Q000502:physiology; D000818:Animals; D003432:Cross-Linking Reagents; D008274:Magnesium / Q000502:physiology; D008954:Models, Biological; D009126:Muscle Relaxation / Q000502:physiology*; D009132:Muscles / Q000502:physiology* / Q000648:ultrastructure*; D009218:Myosins / Q000502:physiology* / Q000648:ultrastructure; D011312:Pressure; D011892:Rana catesbeiana / Q000502:physiology; D012518:Sarcomeres / Q000502:physiology; D013314:Stress, Mechanical; D013329:Structure-Activity Relationship; D014961:X-Ray Diffraction
D003432:Cross-Linking Reagents; D000255:Adenosine Triphosphate; D009218:Myosins; D008274:Magnesium