Tilting of the light-chain region of myosin during step length changes and active force generation in skeletal muscle

FORCE generation and relative sliding between the myosin and actin filaments in muscle are thought to be caused by tilting of the head region of the myosin crossbridges between the filaments1–3. Structural and spectroscopic experiments have demonstrated segmental flexibility of myosin in muscle4–6, but have not shown a direct linkage between tilting of the myosin heads and either force generation or filament sliding. Here we use fluorescence polarization to detect changes in the orientation of the light-chain region of the head, the part most likely to tilt5,7,8, and synchronized head movements by imposing rapid length steps9–11. We found that the light-chain region of the myosin head tilts both during the imposed filament sliding and during the subsequent quick force recovery that is thought to signal the elementary force-generating event.

[1]  G. Piazzesi,et al.  Elastic distortion of myosin heads and repriming of the working stroke in muscle , 1995, Nature.

[2]  K. Holmes,et al.  Induced Changes in Orientation of the Cross-Bridges of Glycerinated Insect Flight Muscle , 1965, Nature.

[3]  Y. Goldman,et al.  Suppression of muscle contraction by vanadate. Mechanical and ligand binding studies on glycerol-extracted rabbit fibers , 1985, The Journal of general physiology.

[4]  R. Mendelson,et al.  Polarization from a helix of fluorophores and its relation to that obtained from muscle. , 1975, Biophysical journal.

[5]  D. D. Thomas,et al.  Spectroscopic probes of muscle cross-bridge rotation. , 1987, Annual review of physiology.

[6]  J. Spudich,et al.  Single myosin molecule mechanics: piconewton forces and nanometre steps , 1994, Nature.

[7]  G. Piazzesi,et al.  The contractile response during steady lengthening of stimulated frog muscle fibres. , 1990, The Journal of physiology.

[8]  T. Yanagida,et al.  Single-molecule analysis of the actomyosin motor using nano-manipulation. , 1994, Biochemical and biophysical research communications.

[9]  M J Kushmerick,et al.  Effects of pH on contraction of rabbit fast and slow skeletal muscle fibers. , 1988, Biophysical journal.

[10]  G. Piazzesi,et al.  Rapid regeneration of the actin-myosin power stroke in contracting muscle , 1992, Nature.

[11]  T. Rowe,et al.  Chimeric myosin regulatory light chains identify the subdomain responsible for regulatory function. , 1992, The EMBO journal.

[12]  H. Huxley,et al.  Changes in the X-ray reflections from contracting muscle during rapid mechanical transients and their structural implications. , 1983, Journal of molecular biology.

[13]  E. Taylor,et al.  Mechanism of adenosine triphosphate hydrolysis by actomyosin. , 1971, Biochemistry.

[14]  H E Huxley,et al.  The Mechanism of Muscular Contraction , 1965, Scientific American.

[15]  I. Schlichting,et al.  Structure of the regulatory domain of scallop myosin at 2.8 Ä resolution , 1994, Nature.

[16]  R. Simmons,et al.  Control of sarcomere length in skinned muscle fibres of Rana temporaria during mechanical transients. , 1984, The Journal of physiology.

[17]  H. Higuchi,et al.  Sliding distance between actin and myosin filaments per ATP molecule hydrolysed in skinned muscle fibres , 1991, Nature.

[18]  A. M. Gordon,et al.  Force and stiffness in glycerinated rabbit psoas fibers. Effects of calcium and elevated phosphate , 1992, The Journal of general physiology.

[19]  D. D. Thomas,et al.  Transients in orientation of a fluorescent cross-bridge probe following photolysis of caged nucleotides in skeletal muscle fibres. , 1992, Journal of molecular biology.

[20]  R. Moss,et al.  Physiological effects accompanying the removal of myosin LC2 from skinned skeletal muscle fibers. , 1982, The Journal of biological chemistry.

[21]  R. Cooke,et al.  Orientation of spin labels attached to cross-bridges in contracting muscle fibres , 1982, Nature.

[22]  J. Corrie,et al.  Synthesis and characterisation of pure isomers of iodoacetamidotetramethylrhodamine , 1994 .

[23]  A. Huxley Muscular contraction. Review lecture , 1974 .

[24]  A. Huxley,et al.  Proposed Mechanism of Force Generation in Striated Muscle , 1971, Nature.

[25]  Y. Goldman,et al.  Measurement of sarcomere shortening in skinned fibers from frog muscle by white light diffraction. , 1987, Biophysical journal.

[26]  R A Milligan,et al.  Structure of the actin-myosin complex and its implications for muscle contraction. , 1993, Science.

[27]  R. Cooke,et al.  The mechanism of muscle contraction. , 1986, CRC critical reviews in biochemistry.

[28]  K. Trybus,et al.  Skeletal muscle myosin light chains are essential for physiological speeds of shortening , 1993, Nature.

[29]  G. Piazzesi,et al.  Myosin head movements are synchronous with the elementary force-generating process in muscle , 1992, Nature.