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. 2001 Jan;80(1):371–378. doi: 10.1016/S0006-3495(01)76021-3

Activating efficiency of Ca2+ and cross-bridges as measured by phosphate analog release.

M Yamaguchi 1, S Takemori 1
PMCID: PMC1301240  PMID: 11159409

Abstract

To assess the activating efficiency of Ca2+ and cross-bridges, the release rates of phosphate analogs from skinned fibers were estimated from the recovery of contractility and that of stiffness. Estimations were performed based on the assumptions that contractility was indicative of the population of analog-free myosin heads and that stiffness reflected the population of formed cross-bridges. Aluminofluoride (AlFx) and orthovanadate (Vi) were used as phosphate analogs with mechanically skinned fibers from rabbit psoas muscle. The use of the analogs enabled the functional assessment of activation level in the total absence of ATP. Fibers loaded with the analogs gradually recovered contractility and stiffness in normal plain rigor solution. The addition of Ca2+ to the plain rigor solution significantly accelerated their recovery, whereas ADP had no appreciable effect. ATP plus Ca2+(contracting condition) accelerated the recovery by several tens of times. These results indicate that the cross-bridges formed during contraction have prominent activating efficiency, which is indispensable to attain full activation. A comparison between the activating efficiency evaluated from stiffness and that from contractility suggested that Ca2+ is more potent in accelerating the binding of actin to analog-bound myosin heads whereas cross-bridges mainly accelerate the subsequent analog-releasing step.

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Selected References

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  1. Bremel R. D., Weber A. Cooperation within actin filament in vertebrate skeletal muscle. Nat New Biol. 1972 Jul 26;238(82):97–101. doi: 10.1038/newbio238097a0. [DOI] [PubMed] [Google Scholar]
  2. Chase P. B., Martyn D. A., Hannon J. D. Activation dependence and kinetics of force and stiffness inhibition by aluminiofluoride, a slowly dissociating analogue of inorganic phosphate, in chemically skinned fibres from rabbit psoas muscle. J Muscle Res Cell Motil. 1994 Apr;15(2):119–129. doi: 10.1007/BF00130423. [DOI] [PubMed] [Google Scholar]
  3. Chase P. B., Martyn D. A., Kushmerick M. J., Gordon A. M. Effects of inorganic phosphate analogues on stiffness and unloaded shortening of skinned muscle fibres from rabbit. J Physiol. 1993 Jan;460:231–246. doi: 10.1113/jphysiol.1993.sp019469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dantzig J. A., Goldman Y. E. Suppression of muscle contraction by vanadate. Mechanical and ligand binding studies on glycerol-extracted rabbit fibers. J Gen Physiol. 1985 Sep;86(3):305–327. doi: 10.1085/jgp.86.3.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dantzig J. A., Hibberd M. G., Trentham D. R., Goldman Y. E. Cross-bridge kinetics in the presence of MgADP investigated by photolysis of caged ATP in rabbit psoas muscle fibres. J Physiol. 1991 Jan;432:639–680. doi: 10.1113/jphysiol.1991.sp018405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fukuda N., Fujita H., Fujita T., Ishiwata S. Regulatory roles of MgADP and calcium in tension development of skinned cardiac muscle. J Muscle Res Cell Motil. 1998 Nov;19(8):909–921. doi: 10.1023/a:1005437517287. [DOI] [PubMed] [Google Scholar]
  7. Güth K., Potter J. D. Effect of rigor and cycling cross-bridges on the structure of troponin C and on the Ca2+ affinity of the Ca2+-specific regulatory sites in skinned rabbit psoas fibers. J Biol Chem. 1987 Oct 5;262(28):13627–13635. [PubMed] [Google Scholar]
  8. Horiuti K., Kagawa K. Effects of ADP and low ATP on the Ca(2+)-sensitive transient contraction upon photolysis of caged ATP in rat muscle fibres: a study on the Bremel-Weber type cooperation. J Muscle Res Cell Motil. 1998 Nov;19(8):923–930. doi: 10.1023/a:1005408300024. [DOI] [PubMed] [Google Scholar]
  9. Horiuti K., Yagi N., Takemori S. Mechanical study of rat soleus muscle using caged ATP and X-ray diffraction: high ADP affinity of slow cross-bridges. J Physiol. 1997 Jul 15;502(Pt 2):433–447. doi: 10.1111/j.1469-7793.1997.433bk.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lehrer S. S., Geeves M. A. The muscle thin filament as a classical cooperative/allosteric regulatory system. J Mol Biol. 1998 Apr 17;277(5):1081–1089. doi: 10.1006/jmbi.1998.1654. [DOI] [PubMed] [Google Scholar]
  11. Marston S. B., Rodger C. D., Tregear R. T. Changes in muscle crossbridges when beta, gamma-imido-ATP binds to myosin. J Mol Biol. 1976 Jun 14;104(1):263–276. doi: 10.1016/0022-2836(76)90012-7. [DOI] [PubMed] [Google Scholar]
  12. Maruta S., Henry G. D., Sykes B. D., Ikebe M. Formation of the stable myosin-ADP-aluminum fluoride and myosin-ADP-beryllium fluoride complexes and their analysis using 19F NMR. J Biol Chem. 1993 Apr 5;268(10):7093–7100. [PubMed] [Google Scholar]
  13. Schoenberg M., Eisenberg E. ADP binding to myosin cross-bridges and its effect on the cross-bridge detachment rate constants. J Gen Physiol. 1987 Jun;89(6):905–920. doi: 10.1085/jgp.89.6.905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Shimizu H., Fujita T., Ishiwata S. Regulation of tension development by MgADP and Pi without Ca2+. Role in spontaneous tension oscillation of skeletal muscle. Biophys J. 1992 May;61(5):1087–1098. doi: 10.1016/S0006-3495(92)81918-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Swartz D. R., Moss R. L., Greaser M. L. Calcium alone does not fully activate the thin filament for S1 binding to rigor myofibrils. Biophys J. 1996 Oct;71(4):1891–1904. doi: 10.1016/S0006-3495(96)79388-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Takemori S., Yamaguchi M., Umazume Y. Physiological significance of viscoelastic structures in myoplasm. Adv Biophys. 1996;33:151–157. [PubMed] [Google Scholar]
  17. Takemori S., Yamaguchi M., Yagi N. An X-ray diffraction study on a single frog skinned muscle fiber in the presence of vanadate. J Biochem. 1995 Mar;117(3):603–608. doi: 10.1093/oxfordjournals.jbchem.a124751. [DOI] [PubMed] [Google Scholar]
  18. Takemori S., Yamaguchi M., Yagi N. Effects of adenosine diphosphate on the structure of myosin cross-bridges: an X-ray diffraction study on a single skinned frog muscle fibre. J Muscle Res Cell Motil. 1995 Dec;16(6):571–577. doi: 10.1007/BF00130238. [DOI] [PubMed] [Google Scholar]
  19. Thirlwell H., Corrie J. E., Reid G. P., Trentham D. R., Ferenczi M. A. Kinetics of relaxation from rigor of permeabilized fast-twitch skeletal fibers from the rabbit using a novel caged ATP and apyrase. Biophys J. 1994 Dec;67(6):2436–2447. doi: 10.1016/S0006-3495(94)80730-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Vibert P., Craig R., Lehman W. Steric-model for activation of muscle thin filaments. J Mol Biol. 1997 Feb 14;266(1):8–14. doi: 10.1006/jmbi.1996.0800. [DOI] [PubMed] [Google Scholar]
  21. Yamaguchi M. Modulating factors of calcium-free contraction at low [MgATP]: a physiological study on the steady states of skinned fibres of frog skeletal muscle. J Muscle Res Cell Motil. 1998 Nov;19(8):949–960. doi: 10.1023/a:1005405002095. [DOI] [PubMed] [Google Scholar]

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