Skip to main content

High ionic strength depresses muscle contractility by decreasing both force per cross-bridge and the number of strongly attached cross-bridges

Abstract

An increase in ionic strength (IS) lowers Ca2+ activated tension in muscle fibres, however, its molecular mechanism is not well understood. In this study, we used single rabbit psoas fibres to perform sinusoidal analyses. During Ca2+ activation, the effects of ligands (ATP, Pi, and ADP) at IS ranging 150–300 mM were studied on three rate constants to characterize elementary steps of the cross-bridge cycle. The IS effects were studied because a change in IS modifies the inter- and intra-molecular interactions, hence they may shed light on the molecular mechanisms of force generation. Both the ATP binding affinity (K 1) and the ADP binding affinity (K 0) increased to 2–3x, and the Pi binding affinity (K 5) decreased to 1/2, when IS was raised from 150 to 300 mM. The effect on ATP/ADP can be explained by stereospecific and hydrophobic interaction, and the effect on Pi can be explained by the electrostatic interaction with myosin. The increase in IS increased cross-bridge detachment steps (k 2 and k −4), indicating that electrostatic repulsion promotes these steps. However, IS did not affect attachment steps (k −2 and k 4). Consequently, the equilibrium constant of the detachment step (K 2) increased by ~100 %, and the force generation step (K 4) decreased by ~30 %. These effects together diminished the number of force-generating cross-bridges by 11 %. Force/cross-bridge (T 56) decreased by 26 %, which correlates well with a decrease in the Debye length that limits the ionic atmosphere where ionic interactions take place. We conclude that the major effect of IS is a decrease in force/cross-bridge, but a decrease in the number of force generating cross-bridge also takes place. The stiffness during rigor induction did not change with IS, demonstrating that in-series compliance is not much affected by IS.

This is a preview of subscription content, access via your institution.

Fig. 1
Scheme 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11
Fig. 12
Fig. 13
Fig. 14

References

  1. Andrews MA, Maughan DW, Nosek TM, Godt RE (1991) Ion-specific and general ionic effects on contraction of skinned fast-twitch skeletal muscle from the rabbit. J Gen Physiol 98(6):1105–1125

    CAS  PubMed  Article  Google Scholar 

  2. Borejdo J, Szczesna-Cordary D, Muthu P, Calander N (2010) Familial hypertrophic cardiomyopathy can be characterized by a specific pattern of orientation fluctuations of actin molecules. Biochemistry 49(25):5269–5277

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  3. Candau R, Kawai M (2011) Correlation between cross-bridge kinetics obtained from Trp fluorescence of myofibril suspensions and mechanical studies of single muscle fibers in rabbit psoas. J Muscle Res Cell Motil 32(4–5):315–326

    CAS  PubMed  Article  Google Scholar 

  4. Colombini B, Nocella M, Bagni MA, Griffiths PJ, Cecchi G (2010) Is the cross-bridge stiffness proportional to tension during muscle fiber activation? Biophys J 98(11):2582–2590

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  5. Geeves MA, Fedorov R, Manstein DJ (2005) Molecular mechanism of actomyosin-based motility. Cell Mol Life Sci 62(13):1462–1477

    CAS  PubMed  Article  Google Scholar 

  6. Geeves MA, Holmes KC (1999) Structural mechanism of muscle contraction. Annu Rev Biochem 68:687–728

  7. Godt RE, Maughan DW (1988) On the composition of the cytosol of relaxed skeletal muscle of the frog. Am J Physiol 254(5 Pt 1):C591–C604

    CAS  PubMed  Google Scholar 

  8. Gordon AM, Godt RE, Donaldson SK, Harris CE (1973) Tension in skinned frog muscle fibers in solutions of varying ionic strength and neutral salt composition. J Gen Physiol 62(5):550–574

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  9. Gulati J, Podolsky RJ (1978) Contraction transients of skinned muscle fibers: effects of calcium and ionic strength. J Gen Physiol 72(5):701–715

    CAS  PubMed  Article  Google Scholar 

  10. Gulati J, Podolsky RJ (1981) Isotonic contraction of skinned muscle fibers on a slow time base: effects of ionic strength and calcium. J Gen Physiol 78(3):233–257

    CAS  PubMed  Article  Google Scholar 

  11. Hanson J, Huxley HE (1957) Quantitative studies on the structure of cross-striated myofibrils. II. Investigations by biochemical techniques. Biochim Biophys Acta 23(2):250–260

    CAS  PubMed  Article  Google Scholar 

  12. Heinl P, Kuhn HJ, Ruegg JC (1974) Tension responses to quick length changes of glycerinated skeletal muscle fibres from the frog and tortoise. J Physiol 237(2):243–258

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  13. Highsmith S, Polosukhina K, Eden D (2000) Myosin motor domain lever arm rotation is coupled to ATP hydrolysis. Biochemistry 39(40):12330–12335

    CAS  PubMed  Article  Google Scholar 

  14. Huxley AF (1974) Muscular contraction. J Physiol 243(1):1–43

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  15. Iwamoto H (2000) Influence of ionic strength on the actomyosin reaction steps in contracting skeletal muscle fibers. Biophys J 78(6):3138–3149

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  16. Kawai M (1982) Correlation between exponential processes and cross-bridge kinetics. Soc Gen Physiol Ser 37:109–130

    CAS  PubMed  Google Scholar 

  17. Kawai M (1986) The role of orthophosphate in crossbridge kinetics in chemically skinned rabbit psoas fibres as detected with sinusoidal and step length alterations. J Muscle Res Cell Motil 7(5):421–434

    CAS  PubMed  Article  Google Scholar 

  18. Kawai M, Brandt PW (1980) Sinusoidal analysis: a high resolution method for correlating biochemical reactions with physiological processes in activated skeletal muscles of rabbit, frog and crayfish. J Muscle Res Cell Motil 1(3):279–303

    CAS  PubMed  Article  Google Scholar 

  19. Kawai M, Halvorson HR (1989) Role of MgATP and MgADP in the cross-bridge kinetics in chemically skinned rabbit psoas fibers. Study of a fast exponential process (C). Biophys J 55(4):595–603

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  20. Kawai M, Halvorson HR (1991) Two step mechanism of phosphate release and the mechanism of force generation in chemically skinned fibers of rabbit psoas muscle. Biophys J 59(2):329–342

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  21. Kawai M, Zhao Y (1993) Cross-bridge scheme and force per cross-bridge state in skinned rabbit psoas muscle fibers. Biophys J 65(2):638–651

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  22. Kawai M, Wray JS, Guth K (1990) Effect of ionic strength on crossbridge kinetics as studied by sinusoidal analysis, ATP hydrolysis rate and X-ray diffraction techniques in chemically skinned rabbit psoas fibres. J Muscle Res Cell Motil 11(5):392–402

    CAS  PubMed  Article  Google Scholar 

  23. Kawai M, Kido T, Vogel M, Fink RH, Ishiwata S (2006) Temperature change does not affect force between regulated actin filaments and heavy meromyosin in single-molecule experiments. J Physiol 574(Pt 3):877–887

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  24. Kerrick WG, Kazmierczak K, Xu Y, Wang Y, Szczesna-Cordary D (2009) Malignant familial hypertrophic cardiomyopathy D166 V mutation in the ventricular myosin regulatory light chain causes profound effects in skinned and intact papillary muscle fibers from transgenic mice. FASEB J 23(3):855–865

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  25. Lobb RR, Stokes AM, Hill HA, Riordan JF (1975) Arginine as the C-1 phosphate binding site in rabbit muscle aldolase. FEBS Lett 54(1):70–72

    CAS  PubMed  Article  Google Scholar 

  26. Maughan DW, Godt RE (1980) A quantitative analysis of elastic, entropic, electrostatic, and osmotic forces within relaxed skinned muscle fibers. Biophys Struct Mech 7(1):17–40

    CAS  PubMed  Article  Google Scholar 

  27. Mettikolla P, Calander N, Luchowski R, Gryczynski I, Gryczynski Z, Zhao J, Szczesna-Cordary D, Borejdo J (2011) Cross-bridge kinetics in myofibrils containing familial hypertrophic cardiomyopathy R58Q mutation in the regulatory light chain of myosin. J Theor Biol 284(1):71–81

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  28. Moore WJ (1962) Physical Chemistry, 3rd edn. Prentice Hall, Englewood Cliffs

    Google Scholar 

  29. Seow CY, Ford LE (1993) High ionic strength and low pH detain activated skinned rabbit skeletal muscle crossbridges in a low force state. J Gen Physiol 101(4):487–511

    CAS  PubMed  Article  Google Scholar 

  30. Sugi H, Abe T, Kobayashi T, Chaen S, Ohnuki Y, Saeki Y, Sugiura S (2013) Enhancement of force generated by individual myosin heads in skinned rabbit psoas muscle fibers at low ionic strength. PLoS One 8(5):e63658

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  31. Thames MD, Teichholz LE, Podolsky RJ (1974) Ionic strength and the contraction kinetics of skinned muscle fibers. J Gen Physiol 63(4):509–530

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  32. Wang L, Kawai M (2013) A re-interpretation of the rate of tension redevelopment (k(TR)) in active muscle. J Muscle Res Cell Motil 34(5–6):407–415

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  33. Wang Y, Xu Y, Kerrick WG, Wang Y, Guzman G, Diaz-Perez Z, Szczesna-Cordary D (2006) Prolonged Ca2+ and force transients in myosin RLC transgenic mouse fibers expressing malignant and benign FHC mutations. J Mol Biol 361(2):286–299

    CAS  PubMed  Article  Google Scholar 

  34. Wang L, Muthu P, Szczesna-Cordary D, Kawai M (2013) Diversity and similarity of motor function and cross-bridge kinetics in papillary muscles of transgenic mice carrying myosin regulatory light chain mutations D166 V and R58Q. J Mol Cell Cardiol 62:153–163

    CAS  PubMed  Article  Google Scholar 

  35. Wang L, Ji X, Sadayappan S, Kawai M (2014) Phosphorylation of cMyBP-C affects contractile mechanisms in a site-specific manner. Biophys J 106(5):1112–1122

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  36. Yan Q, Sun Y, Lin J (1996) A quantitative study on the effect of breathing exercises in improving respiratory muscle contration. Zhonghua nei ke za zhi 35(4):235–238

    CAS  PubMed  Google Scholar 

  37. Yount RG, Lawson D, Rayment I (1995) Is myosin a “back door” enzyme? Biophys J 68(4 Suppl):44S–47S; discussion 47S–49S

  38. Zhao Y, Kawai M (1994) Kinetic and thermodynamic studies of the cross-bridge cycle in rabbit psoas muscle fibers. Biophys J 67(4):1655–1668

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  39. Zhao Y, Kawai M (1996) Inotropic agent EMD-53998 weakens nucleotide and phosphate binding to cross bridges in porcine myocardium. Am J Physiol 271(4 Pt 2):H1394–H1406

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the National Institutes of Health HL070041 (MK), and The American Heart Association 13GRNT16810043 (MK). This study was carried out during Dr. Anzel Bahadir’s visit to The University of Iowa with a scholarship (82444403-299-1926) funded by Higher Educational Council of Turkey. The content is solely the responsibility of the authors and does not necessarily reflect the official views of the funding organizations.

Conflict of interest

The authors have no conflicts of interests.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Masataka Kawai.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Wang, L., Bahadir, A. & Kawai, M. High ionic strength depresses muscle contractility by decreasing both force per cross-bridge and the number of strongly attached cross-bridges. J Muscle Res Cell Motil 36, 227–241 (2015). https://doi.org/10.1007/s10974-015-9412-6

Download citation

Keywords

  • Kinetics
  • Elementary steps
  • Debye length
  • Ionic atmosphere
  • Sinusoidal analysis
  • Rabbit psoas fibres