Abstract
The temporal relationship between myosin phosphorylation, contractile force and ATPase activity was studied in skinned preparations from the guinea-pig Taenia coli. When free Calcium concentration ([Ca2+]) was increased from pCa (−log[Ca2+]) 9 to pCa 4.5 at low calmodulin concentration (0.05 μM), ATPase activity and myosin light-chain phosphorylation rose quickly, while the increase in force and stiffness was delayed. The time-course of tension increase was faster at higher calmodulin concentrations (5 μM), although the maximal level of phosphorylation was unchanged. Lowering the calcium concentration from pCa 4.5 to pCa 9 at the plateau of contraction caused a rapid decrease in ATPase activity and in myosin phosphorylation, while force and stiffness decayed more slowly. The force decay could be accelerated by inorganic phosphate. These results suggest that, during contraction, force may be produced actively by phosphorylated and ATP-splitting cross-bridges, but may be maintained by dephosphorylated cross-bridges which cycle slowly. However, force could also be modulated by calmodulin and inorganic phosphate in a manner not involving an alteration in the extent of myosin phosphorylation.
Similar content being viewed by others
References
Arheden H, Arner A, Hellstrand P (1988) Cross-bridge behaviour in skinned smooth muscle of the guinea pig taenia coli at altered ionic strength. J Physiol (Lond) 403: 539–558
Arner A (1982) Mechanical characteristics of chemically skinned guinea pig Taenia coli. Pflügers Arch 395: 277–284
Arner A, Goody RS, Rapp G, Rüegg JC (1987) Relaxation of chemically skinned guinea pig taenia coli smooth muscle from rigor by photolytic release of adenosine-5′-triphosphate. J Muscle Res Cell Motil 8: 377–385
Brenner B (1988) Effect of Ca2+ on cross-bridge turnover kinetics in single rabbit psoas fibres: implications for regulation of muscle contraction. Proc Natl Acad Sci USA 85: 3265–3269
Chacko S, Conti MA, Adelstein RS (1977) Effect of phosphorylation of smooth muscle myosin on actin activation and Ca2+ regulation. Proc Natl Acad Sci USA 74: 129–133
Chatterjee M, Murphy RA (1983) Calcium-dependent stress maintenance without myosin phosphorylation in skinned smooth muscle. Science 221: 464–466
Dillon PF, Aksoy MO, Driska SP, Murphy RA (1981) Myosin phosphorylation and the cross-bridge cycle in arterial smooth muscle. Science 211: 495–497
Driska SP, Aksoy MO, Murphy RA (1981) Myosin light chain phosphorylation associated with contraction in arterial smooth muscle. Am J Physiol 240: C222-C233
Fabiato A (1981) Myoplasmic free calcium concentration reached during twitch of an intact isolated cardiac cell and during calcium-induced release of calcium from the sarcoplasmatic reticulum of a skinned cardiac cell from the adult rat or rabbit ventricle. J Gen Physiol 78: 457–497
Fabiato A, Fabiato A (1979) Calculator programs for computing the composition of the solutions containing multiple metals and ligands used for experiments in skinned muscle cells. J Physiol (Paris) 75: 463–505
Ford LE, Huxley AF, Simmons RM (1977) Tension responses to sudden length change in stimulated frog muscle fibres near slack length. J Physiol (Lond) 269: 441–515
Gagelmann M, Güth K (1987) Effect of inorganic phosphate on the Ca2+ sensitivity in skinned taenia coli smooth muscle fibres. Comparison of tension, ATPase activity, and phosphorylation of the regulatory myosin light chains. Biophys J 51: 457–463
Güth K, Junge J (1982) Low Ca2+ impedes cross-bridge detachment in chemically skinned Taenia coli. Nature 300: 775–776
Güth K, Mrwa U (1985) A slow calcium-dependent process precedes force development, phosphorylation and ATPase activity in skinned taenia coli. J Muscle Res Cell Motil 6:116
Güth K, Wojciechowski R (1986) Perfusion cuvette for simultaneous measurement of mechanical, optical and energetic parameters of skinned fibres. Pflügers Arch 407: 552–557
Güth K, Kuhn HJ, Drexler B, Berberich W, Rüegg JC (1979) Stiffness and tension during and after sudden length changes of glycerinated single insect fibrillar muscle fibres. Biophys Struct Mech 5: 255–276
Hai C-M, Murphy RA (1988) Cross-bridge phosphorylation and regulation of latch state in smooth muscle. Am J Physiol 254: C99-C106
Hartshorne DJ, Siemankowsky RF (1981) Regulation of smooth muscle actomyosin. Annu Rev Physiol 43: 519–530
Hellstrand P, Arner A (1985) Myosin light chain phosphorylation and the cross-bridge cycle at low substrate concentration in chemically skinned guinea pig taenia coli. Pflügers Arch 405: 323–328
Huxley AF (1957) Muscle structure and theories of muscle contraction. Prog Biophys Biophys Chem 7: 255–318
Kühn H, Tewes A, Gagelmann M, Güth K, Arner A, Rüegg JC (1985) Temporal relationship of force, ATPase activity and myosin light chain phosphorylation during contraction and relaxation in skinned guinea pig taenia coli. Pflügers Arch 505 [Suppl 2]: R61
Marston SB, Smith CWJ (1985) The thin filaments of smooth muscles. J Muscle Res Cell Motil 6: 669–708
Ngai PK, Walsh MP (1984) Inhibition of smooth muscle actin-activated myosin Mg2+-ATPase activity by caldesmon. J Biol Chem 259: 13656–13659
Ritchie JM (1954) The effect of nitrate on the active state of muscle. J Physiol (Lond) 126: 155–168
Sobue K, Muramoto Y, Fujita M, Kakiuchi S (1981) Purification of a calmodulin-binding protein from chicken gizzard that interacts with F-actin. Proc Natl Acad Sci USA 78: 5652–5655
Sparrow MP, Mrwa U, Hofmann F, Rüegg JC (1981) Calmodulin is essential for smooth muscle contraction. FEBS Lett 125: 141–145
Wagner J, Rüegg JC (1986) Skinned smooth muscle: calcium-calmodulin activation independent of myosin phosphorylation. Pflügers Arch 407: 569–571
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kühn, H., Tewes, A., Gagelmann, M. et al. Temporal relationship between force, ATPase activity, and myosin phosphorylation during a contraction/relaxation cycle in a skinned smooth muscle. Pflügers Arch. 416, 512–518 (1990). https://doi.org/10.1007/BF00382683
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00382683