Advertisement

Cellular and Molecular Bioengineering

, Volume 6, Issue 2, pp 183–198 | Cite as

Slowed Dynamics of Thin Filament Regulatory Units Reduces Ca2+-Sensitivity of Cardiac Biomechanical Function

  • Campion K. P. Loong
  • Aya K. Takeda
  • Myriam A. Badr
  • Jordan S. Rogers
  • P. Bryant ChaseEmail author
Article

Abstract

Actomyosin kinetics in both skinned skeletal muscle fibers at maximum Ca2+-activation and unregulated in vitro motility assays are modulated by solvent microviscosity in a manner consistent with a diffusion limited process. Viscosity might also influence cardiac thin filament Ca2+-regulatory protein dynamics. In vitro motility assays were conducted using thin filaments reconstituted with recombinant human cardiac troponin and tropomyosin; solvent microviscosity was varied by addition of sucrose or glucose. At saturating Ca2+, filament sliding speed (s) was inversely proportional to viscosity. Ca2+-sensitivity (pCa 50) of s decreased markedly with elevated viscosity (η/η 0 ≥ ~1.3). For comparison with unloaded motility assays, steady-state isometric force (F) and kinetics of isometric tension redevelopment (k TR) were measured in single, permeabilized porcine cardiomyocytes when viscosity surrounding the myofilaments was altered. Maximum Ca2+-activated F changed little for sucrose ≤0.3 M (η/η 0 ~ 1.4) or glucose ≤0.875 M (η/η 0 ~ 1.66), but decreased at higher concentrations. Sucrose (0.3 M) or glucose (0.875 M) decreased pCa 50 for F. k TR at saturating Ca2+ decreased steeply and monotonically with increased viscosity but there was little effect on k TR at sub-maximum Ca2+. Modeling indicates that increased solutes affect dynamics of cardiac muscle Ca2+-regulatory proteins to a much greater extent than actomyosin cross-bridge cycling.

Keywords

Skinned myocyte Myosin Actin Troponin Tropomyosin In vitro motility assay Kinetics of isometric tension redevelopment Microviscosity Monosaccharide glucose Disaccharide sucrose 

Notes

Acknowledgments

This work was supported by National Institute of Health grants HL63974 (PBC), a Florida State University Center for Materials Research and Technology (MARTECH) Pre-doctoral Fellowship (MAB), and American Heart Association Pre-doctoral Fellowships 0615164B (AKT) and 0815127E (CKPL). We thank Bradley’s Country Store, Tallahassee, FL, for generously supplying porcine hearts.

Conflict of interest

The authors have no conflicts of interest to declare.

References

  1. 1.
    Bai, F., A. Weis, A. K. Takeda, P. B. Chase, and M. Kawai. Enhanced active cross-bridges during diastole: molecular pathogenesis of tropomyosin’s HCM mutations. Biophys. J. 100:1014–1023, 2011.CrossRefGoogle Scholar
  2. 2.
    Brenner, B. Technique for stabilizing the striation pattern in maximally calcium-activated skinned rabbit psoas fibers. Biophys. J. 41:99–102, 1983.CrossRefGoogle Scholar
  3. 3.
    Brenner, B. Effect of Ca2+ on cross-bridge turnover kinetics in skinned single rabbit psoas fibers: implications for regulation of muscle contraction. Proc. Natl. Acad. Sci. U.S.A. 85:3265–3269, 1988.CrossRefGoogle Scholar
  4. 4.
    Brenner, B., and E. Eisenberg. Rate of force generation in muscle: correlation with actomyosin ATPase activity in solution. Proc. Natl. Acad. Sci. U.S.A. 83:3542–3546, 1986.CrossRefGoogle Scholar
  5. 5.
    Brunet, N. M., G. Mihajlović, K. Aledealat, F. Wang, P. Xiong, S. von Molnár, and P. B. Chase. Micromechanical thermal assays of Ca2+-regulated thin-filament function and modulation by hypertrophic cardiomyopathy mutants of human cardiac troponin. J. Biomed. Biotechnol. 2012:657523, 2012.CrossRefGoogle Scholar
  6. 6.
    Campbell, K. Rate constant of muscle force redevelopment reflects cooperative activation as well as cross-bridge kinetics. Biophys. J. 72:254–262, 1997.CrossRefGoogle Scholar
  7. 7.
    Cecchi, G., M. A. Bagni, P. J. Griffiths, C. C. Ashley, and Y. Maeda. Detection of radial crossbridge force by lattice spacing changes in intact single muscle fibers. Science 250:1409–1411, 1990.CrossRefGoogle Scholar
  8. 8.
    Cellmer, T., E. R. Henry, J. Hofrichter, and W. A. Eaton. Measuring internal friction of an ultrafast-folding protein. Proc. Natl. Acad. Sci. U.S.A. 105:18320–18325, 2008.CrossRefGoogle Scholar
  9. 9.
    Chase, P. B., Y. Chen, K. Kulin, and T. L. Daniel. Viscosity and solute dependence of F-actin translocation by rabbit skeletal heavy meromyosin. Am. J. Physiol. Cell Physiol. 278:C1088–C1098, 2000.Google Scholar
  10. 10.
    Chase, P. B., T. M. Denkinger, and M. J. Kushmerick. Effect of viscosity on mechanics of single, skinned fibers from rabbit psoas muscle. Biophys. J. 74:1428–1438, 1998.CrossRefGoogle Scholar
  11. 11.
    Chase, P. B., D. A. Martyn, and J. D. Hannon. Isometric force redevelopment of skinned muscle fibers from rabbit with and without Ca2+. Biophys. J. 67:1994–2001, 1994.CrossRefGoogle Scholar
  12. 12.
    Édes, I. F., D. Czuriga, G. Csányi, S. Chłopicki, F. A. Recchia, A. Borbély, Z. Galajda, I. Édes, J. van der Velden, G. J. M. Stienen, and Z. Papp. Rate of tension redevelopment is not modulated by sarcomere length in permeabilized human, murine, and porcine cardiomyocytes. Am. J. Physiol. Regul. Integr. Comp. Physiol. 293:R20–R29, 2007.CrossRefGoogle Scholar
  13. 13.
    Endo, M., T. Kitazawa, M. Iino, and Y. Kakuta. Effect of “viscosity” of the medium on mechanical properties of skinned skeletal muscle fibers. In: Cross-Bridge Mechanism in Muscle Contraction, edited by H. Sugi, and G. H. Pollack. Tokyo: University of Tokyo Press, 1979, pp. 365–374.Google Scholar
  14. 14.
    Fajer, P. Conformational switching in muscle. Adv. Exp. Med. Biol. 547:61–80, 2004.CrossRefGoogle Scholar
  15. 15.
    Gafurov, B., S. Fredricksen, A. Cai, B. Brenner, P. B. Chase, and J. M. Chalovich. The Δ14 mutant of troponin T enhances ATPase activity and alters the cooperative binding of S1-ADP to regulated actin. Biochemistry 43:15276–15285, 2004.CrossRefGoogle Scholar
  16. 16.
    Gillis, T. E., D. A. Martyn, A. J. Rivera, and M. Regnier. Investigation of thin filament near-neighbour regulatory unit interactions during force development in skinned cardiac and skeletal muscle. J. Physiol. 580:561–576, 2007.CrossRefGoogle Scholar
  17. 17.
    Gordon, A. M., Y. Chen, B. Liang, M. LaMadrid, Z. Luo, and P. B. Chase. Skeletal muscle regulatory proteins enhance F-actin in vitro motility. Adv. Exp. Med. Biol. 453:187–197, 1998.CrossRefGoogle Scholar
  18. 18.
    Gordon, A. M., E. Homsher, and M. Regnier. Regulation of contraction in striated muscle. Physiol. Rev. 80:853–924, 2000.Google Scholar
  19. 19.
    Gordon, A. M., M. LaMadrid, Y. Chen, Z. Luo, and P. B. Chase. Calcium regulation of skeletal muscle thin filament motility in vitro. Biophys. J. 72:1295–1307, 1997.CrossRefGoogle Scholar
  20. 20.
    Hancock, W. O., L. L. Huntsman, and A. M. Gordon. Models of calcium activation account for differences between skeletal and cardiac force redevelopment kinetics. J. Muscle Res. Cell Motil. 18:671–681, 1997.CrossRefGoogle Scholar
  21. 21.
    Hannon, J. D., P. B. Chase, D. A. Martyn, L. L. Huntsman, M. J. Kushmerick, and A. M. Gordon. Calcium-independent activation of skeletal muscle fibers by a modified form of cardiac troponin C. Biophys. J. 64:1632–1637, 1993.CrossRefGoogle Scholar
  22. 22.
    Hinken, A. C., F. S. Korte, and K. S. McDonald. Porcine cardiac myocyte power output is increased after chronic exercise training. J. Appl. Physiol. 101:40–46, 2006.CrossRefGoogle Scholar
  23. 23.
    Homsher, E., F. Wang, and J. R. Sellers. Factors affecting movement of F-actin filaments propelled by skeletal muscle heavy meromyosin. Am. J. Physiol. 262:C714–C723, 1992.Google Scholar
  24. 24.
    Köhler, J., Y. Chen, B. Brenner, A. M. Gordon, T. Kraft, D. A. Martyn, M. Regnier, A. J. Rivera, C.-K. Wang, and P. B. Chase. Familial hypertrophic cardiomyopathy mutations in troponin I (K183Δ, G203S, K206Q) enhance filament sliding. Physiol. Genomics 14:117–128, 2003.Google Scholar
  25. 25.
    Kron, S. J., Y. Y. Toyoshima, T. Q. P. Uyeda, and J. A. Spudich. Assays for actin sliding movement over myosin-coated surfaces. Methods Enzymol. 196:399–416, 1991.CrossRefGoogle Scholar
  26. 26.
    Kumar, R., and A. K. Bhuyan. Viscosity scaling for the glassy phase of protein folding. J. Phys. Chem. B 112:12549–12554, 2008.CrossRefGoogle Scholar
  27. 27.
    Lamb, G. D., D. G. Stephenson, and G. J. Stienen. Effects of osmolality and ionic strength on the mechanism of Ca2+ release in skinned skeletal muscle fibres of the toad. J. Physiol. 464:629–648, 1993.Google Scholar
  28. 28.
    Landesberg, A., and S. Sideman. Coupling calcium binding to troponin C and cross-bridge cycling in skinned cardiac cells. Am. J. Physiol. 266:H1260–H1271, 1994.Google Scholar
  29. 29.
    Li, H.-C., K. Hideg, and P. G. Fajer. The mobility of troponin C and troponin I in muscle. J. Mol. Recognit. 10:194–201, 1997.CrossRefGoogle Scholar
  30. 30.
    Liang, B., Y. Chen, C.-K. Wang, Z. Luo, M. Regnier, A. M. Gordon, and P. B. Chase. Ca2+ regulation of rabbit skeletal muscle thin filament sliding: role of cross-bridge number. Biophys. J. 85:1775–1786, 2003.CrossRefGoogle Scholar
  31. 31.
    Loong, C. K. P., M. A. Badr, and P. B. Chase. Tropomyosin flexural rigidity and single Ca2+ regulatory unit dynamics: implications for cooperative regulation of cardiac muscle contraction and cardiomyocyte hypertrophy. Front. Physiol. 3:80, 2012.CrossRefGoogle Scholar
  32. 32.
    Loong, C. K. P., H.-X. Zhou, and P. B. Chase. Familial hypertrophic cardiomyopathy related E180G mutation increases flexibility of human cardiac α-tropomyosin. FEBS Lett. 586:3503–3507, 2012.CrossRefGoogle Scholar
  33. 33.
    Margossian, S. S., and S. Lowey. Preparation of myosin and its subfragments from rabbit skeletal muscle. Methods Enzymol. 85:55–71, 1982.CrossRefGoogle Scholar
  34. 34.
    Martyn, D. A., B. B. Adhikari, M. Regnier, J. Gu, S. Xu, and L. C. Yu. Response of equatorial x-ray reflections and stiffness to altered sarcomere length and myofilament lattice spacing in relaxed skinned cardiac muscle. Biophys. J. 86:1002–1011, 2004.CrossRefGoogle Scholar
  35. 35.
    Mathur, M. C., P. B. Chase, and J. M. Chalovich. Several cardiomyopathy causing mutations on tropomyosin either destabilize the active state of actomyosin or alter the binding properties of tropomyosin. Biochem. Biophys. Res. Commun. 406:74–78, 2011.CrossRefGoogle Scholar
  36. 36.
    Maytum, R., S. S. Lehrer, and M. A. Geeves. Cooperativity and switching within the three-state model of muscle regulation. Biochemistry 38:1102–1110, 1999.CrossRefGoogle Scholar
  37. 37.
    McDonald, K. S., and R. L. Moss. Osmotic compression of single cardiac myocytes eliminates the reduction in Ca2+ sensitivity of tension at short sarcomere length. Circ. Res. 77:199–205, 1995.CrossRefGoogle Scholar
  38. 38.
    McDonald, K. S., M. R. Wolff, and R. L. Moss. Force–velocity and power–load curves in rat skinned cardiac myocytes. J. Physiol. 511:519–531, 1998.CrossRefGoogle Scholar
  39. 39.
    McKie, J. E., and J. F. Brandts. High precision capillary viscometry. Methods Enzymol. 26:257–288, 1972.CrossRefGoogle Scholar
  40. 40.
    Metzger, J. M., and R. L. Moss. Calcium-sensitive cross-bridge transitions in mammalian fast and slow skeletal muscle fibers. Science 247:1088–1090, 1990.CrossRefGoogle Scholar
  41. 41.
    Miller-Jaster, K. N., C. E. P. Aronin, and W. H. Guilford. A quantitative comparison of blocking agents in the in vitro motility assay. Cell. Mol. Bioeng. 5:44–51, 2012.CrossRefGoogle Scholar
  42. 42.
    Moreno-Gonzalez, A., T. E. Gilles, A. J. Rivera, P. B. Chase, D. A. Martyn, and M. Regnier. Thin filament regulation of force redevelopment kinetics in rabbit skeletal muscle fibres. J. Physiol. 579:313–326, 2007.CrossRefGoogle Scholar
  43. 43.
    Pardee, J. D., and J. A. Spudich. Purification of muscle actin. Methods Enzymol. 85:164–181, 1982.CrossRefGoogle Scholar
  44. 44.
    Parmacek, M. S., and R. J. Solaro. Biology of the troponin complex in cardiac myocytes. Prog. Cardiovasc. Dis. 47:159–176, 2004.CrossRefGoogle Scholar
  45. 45.
    Regnier, M., D. A. Martyn, and P. B. Chase. Calmidazolium alters Ca2+ regulation of tension redevelopment rate in skinned skeletal muscle. Biophys. J. 71:2786–2794, 1996.CrossRefGoogle Scholar
  46. 46.
    Regnier, M., D. A. Martyn, and P. B. Chase. Calcium regulation of tension redevelopment kinetics with 2-deoxy-ATP or low [ATP] in rabbit skeletal muscle. Biophys. J. 74:2005–2015, 1998.CrossRefGoogle Scholar
  47. 47.
    Regnier, M., A. J. Rivera, M. A. Bates, C.-K. Wang, P. B. Chase, and A. M. Gordon. Thin filament near-neighbor regulatory unit interactions affect rabbit skeletal muscle steady state force–Ca2+ relations. J. Physiol. 540:485–497, 2002.CrossRefGoogle Scholar
  48. 48.
    Regnier, M., A. J. Rivera, P. B. Chase, L. B. Smillie, and M. M. Sorenson. Regulation of skeletal muscle tension redevelopment by troponin C constructs with different Ca2+ affinities. Biophys. J. 76:2664–2672, 1999.CrossRefGoogle Scholar
  49. 49.
    Rice, J. J., G. Stolovitzky, Y. Tu, and P. P. de Tombe. Ising model of cardiac thin filament activation with nearest-neighbor cooperative interactions. Biophys. J. 84:897–909, 2003.CrossRefGoogle Scholar
  50. 50.
    Schiaffino, S., and C. Reggiani. Molecular diversity of myofibrillar proteins: gene regulation and functional significance. Physiol. Rev. 76:371–423, 1996.Google Scholar
  51. 51.
    Schoffstall, B., N. M. Brunet, F. Wang, S. Williams, A. T. Barnes, V. F. Miller, L. A. Compton, L. A. McFadden, D. W. Taylor, R. Dhanarajan, M. Seavy, and P. B. Chase. Ca2+-sensitivity of regulated cardiac thin filament sliding does not depend on myosin isoform. J. Physiol. 577:935–944, 2006.CrossRefGoogle Scholar
  52. 52.
    Schoffstall, B., A. Clark, and P. B. Chase. Positive inotropic effects of low dATP/ATP ratios on mechanics and kinetics of porcine cardiac muscle. Biophys. J. 91:2216–2226, 2006.CrossRefGoogle Scholar
  53. 53.
    Schoffstall, B., A. Kataoka, A. Clark, and P. B. Chase. Effects of rapamycin on cardiac and skeletal muscle contraction and crossbridge cycling. J. Pharmacol. Exp. Ther. 312:12–18, 2005.CrossRefGoogle Scholar
  54. 54.
    Schoffstall, B., V. A. LaBarbera, N. M. Brunet, B. J. Gavino, L. Herring, S. Heshmati, B. H. Kraft, V. Inchausti, N. L. Meyer, D. Moonoo, A. K. Takeda, and P. B. Chase. Interaction between troponin and myosin enhances contractile activity of myosin in cardiac muscle. DNA Cell Biol. 30:653–659, 2011.CrossRefGoogle Scholar
  55. 55.
    Sweeney, H. L., and J. T. Stull. Alteration of cross-bridge kinetics by myosin light chain phosphorylation in rabbit skeletal muscle: implications for regulation of actin-myosin interaction. Proc. Natl. Acad. Sci. U.S.A. 87:414–418, 1990.CrossRefGoogle Scholar
  56. 56.
    Szczesna, D., and P. G. Fajer. The tropomyosin domain is flexible and disordered in reconstituted thin filaments. Biochemistry 34:3614–3620, 1995.CrossRefGoogle Scholar
  57. 57.
    Tikunova, S. B., and J. P. Davis. Designing calcium-sensitizing mutations in the regulatory domain of cardiac troponin C. J. Biol. Chem. 279:35341–35352, 2004.CrossRefGoogle Scholar
  58. 58.
    Wang, F., N. M. Brunet, J. R. Grubich, E. Bienkiewicz, T. M. Asbury, L. A. Compton, G. Mihajlović, V. F. Miller, and P. B. Chase. Facilitated cross-bridge interactions with thin filaments by familial hypertrophic cardiomyopathy mutations in α-tropomyosin. J. Biomed. Biotechnol. 2011:435271, 2011.Google Scholar
  59. 59.
    Wang, Y., and F. Fuchs. Interfilament spacing, Ca2+ sensitivity, and Ca2+ binding in skinned bovine cardiac muscle. J. Muscle Res. Cell Motil. 22:251–257, 2001.zbMATHCrossRefGoogle Scholar
  60. 60.
    Washio, T., J. I. Okada, S. Sugiura, and T. Hisada. Approximation for cooperative interactions of a spatially-detailed cardiac sarcomere model. Cell. Mol. Bioeng. 5:113–126, 2012.CrossRefGoogle Scholar
  61. 61.
    Weast, R. C. (ed.). CRC Handbook of Chemistry and Physics. Boca Raton, FL: CRC Press, 1982.Google Scholar

Copyright information

© Biomedical Engineering Society 2013

Authors and Affiliations

  • Campion K. P. Loong
    • 1
    • 2
    • 4
  • Aya K. Takeda
    • 1
    • 3
    • 5
  • Myriam A. Badr
    • 1
    • 3
  • Jordan S. Rogers
    • 1
    • 6
  • P. Bryant Chase
    • 1
    Email author
  1. 1.Department of Biological ScienceThe Florida State UniversityTallahasseeUSA
  2. 2.Department of PhysicsThe Florida State UniversityTallahasseeUSA
  3. 3.Program in Molecular BiophysicsThe Florida State UniversityTallahasseeUSA
  4. 4.MathWorks, Inc.NatickUSA
  5. 5.Genaris, Inc.YokohamaJapan
  6. 6.Shands HospitalUniversity of Florida College of MedicineGainesvilleUSA

Personalised recommendations