Cryo-atomic force microscopy of smooth muscle myosin.
The motor and regulatory domains of the head and the 14-nm pitch of the alpha-helical coiled-coil of the tail of extended (6S) smooth-muscle myosin molecules were imaged with cryo atomic force microscopy at 80-85 K, and the effects of thiophosphorylation of the regulatory light chain were examined. The tail was 4 nm shorter in thiophosphorylated than in nonphosphorylated myosin. The first major bend was invariant, at approximately 51 nm from the head-tail junction (H-T), coincident with low probability in the paircoil score. The second major bend was 100 nm from the H-T junction in nonphosphorylated and closer to a skip residue than the bend (at 95 nm) in thiophosphorylated molecules. The shorter tail and distance between the two major bends induced by thiophosphorylation are interpreted to result from melting of the coiled-coil. An additional bend not previously reported occurred, with a lower frequency, approximately 24 nm from the H-T. The range of separation between the two heads was greater in thiophosphorylated molecules. Occasional high-resolution images showed slight unwinding of the coiled-coil of the base of the heads. We suggest that phosphorylation of MLC20 can affect the structure of extended, 6S myosin.
Zhang
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Y
Department of Molecular Physiology, University of Virginia Health Sciences Center, Charlottesville 22906-0011, USA.
Shao
Z
Z
Somlyo
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AP
Somlyo
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AV
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D005615:Freezing; D005895:Gizzard, Avian; D018625:Microscopy, Atomic Force / Q000379:methods; D009130:Muscle, Smooth / Q000502:physiology*; D009218:Myosins / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure*; D010749:Phosphoprotein Phosphatases; D010766:Phosphorylation; D017433:Protein Structure, Secondary; D014422:Turkeys
C022571:adenosine 5'-O-(3-thiotriphosphate); D000255:Adenosine Triphosphate; D010749:Phosphoprotein Phosphatases; D009218:Myosins