Advertisement

Expression of tropomyosin-κ induces dilated cardiomyopathy and depresses cardiac myofilament tension by mechanisms involving cross-bridge dependent activation and altered tropomyosin phosphorylation

  • Chehade N. Karam
  • Chad M. Warren
  • Sudarsan Rajan
  • Pieter P. de Tombe
  • David F. Wieczorek
  • R. John Solaro
Original Paper

Abstract

Tropomyosin-kappa (TPM1-κ) is a newly discovered tropomyosin (TM) isoform that is exclusively expressed in the human heart and generated by an alternative splicing of the α-TM gene. We reported that TPM1-κ expression was increased in the hearts of patients with chronic dilated cardiomyopathy (DCM). To increase our understanding of the significance of this shift in isoform population, we generated transgenic (TG) mice expressing TPM1-κ in the cardiac compartment where TPM1-κ replaces 90% of the native TM. We previously showed that there was a significant inhibition of the ability of strongly bound cross-bridges to induce activation of TG myofilaments (Rajan et al., Circulation 121:410–418, 2010). Here, we compared the force–Ca2+ relations in detergent extracted (skinned) fiber bundles isolated from non-transgenic (NTG) and TG-TPM1-κ hearts at two sarcomere lengths (SLs). Our data demonstrated a significant decrease in the Ca2+ sensitivity of the myofilaments from TG-TPM1-κ hearts with no change in the maximum developed tension, length-dependent activation, and the ratio of ATPase rate to tension. There was also no difference in the affinity and cooperativity of Ca2+-binding to troponin in thin filaments reconstituted with either TPM1-κ or α-TM. We also compared protein phosphorylation in NTG and TG-TPM1-κ myofilaments. There was a decrease in the total phosphorylation of TPM1-κ compared to α-TM, but no significant change in other major sarcomeric proteins. Our results identify a novel mode of myofilament desensitization to Ca2+ associated with a DCM linked switch in TM isoform population.

Keywords

Calcium sensitivity Tension cost Thin filament regulation ATPase Sarcomeric proteins 

Notes

Acknowledgments

The authors would like to thank both Dr. Tomoyoshi Kobayashi for his help with the TnC binding measurements and Bindiya G. Patel for her assistance with the 2D-DIGE experiments. This work was supported by American Heart Association-Midwest Pre-Doctoral Fellowship (CNK) and by NIH Grants PO1 HL062426 (RJS, PdeT), RO1 HL022231 (RJS), RO1 HL081680, and RO1 HL071952 (DFW).

References

  1. Davis JP, Norman C, Kobayashi T, Solaro RJ, Swartz DR, Tikunova SB (2007) Effects of thin and thick filament proteins on calcium binding and exchange with cardiac troponin C. Biophys J 92(9):3195–3206CrossRefPubMedGoogle Scholar
  2. de Tombe PP, Stienen GJ (1995) Protein kinase A does not alter economy of force maintenance in skinned rat cardiac trabeculae. Circ Res 76(5):734–741PubMedGoogle Scholar
  3. de Tombe PP, Stienen GJ (2007) Impact of temperature on cross-bridge cycling kinetics in rat myocardium. J Physiol 584(Pt 2):591–600CrossRefPubMedGoogle Scholar
  4. Denz CR, Narshi A, Zajdel RW, Dube DK (2004) Expression of a novel cardiac-specific tropomyosin isoform in humans. Biochem Biophys Res Commun 320:1291–1297CrossRefPubMedGoogle Scholar
  5. Fabiato A (1988) Computer programs for calculating total from specified free or free from specified total ionic concentrations in aqueous solutions containing multiple metals and ligands. Methods Enzymol 157:378–417CrossRefPubMedGoogle Scholar
  6. Heeley DH, Watson MH, Mak AS, Dubord P, Smillie LB (1989) Effect of phosphorylation on the interaction and functional properties of rabbit striated muscle αα-tropomyosin. J Biol Chem 264(5):2424–2430PubMedGoogle Scholar
  7. Jagatheesan G, Rajan S, Petrashevskaya N, Schwartz A, Boivin G, Arteaga GM, Solaro RJ, Liggett SB, Wieczorek DF (2007) Rescue of tropomyosin-induced familial hypertrophic cardiomyopathy mice by transgenesis. Am J Physiol Heart Circ Physiol 293(2):H949–H958CrossRefPubMedGoogle Scholar
  8. Kobayashi T, Solaro RJ (2006) Increased Ca2+ affinity of cardiac thin filaments reconstituted with cardiomyopathy-related mutant cardiac troponin I. J Biol Chem 281(19):13471–13477CrossRefPubMedGoogle Scholar
  9. Kobayashi T, Zhao X, Wade R, Collins JH (1999) Ca-dependent interaction of the inhibitory region of troponin I with acidic residues in the N-terminal domain of troponin C. Biochim Biophys Acta 1430:214–221CrossRefPubMedGoogle Scholar
  10. Lakdawala NK, Dellefave L, Redwood CS, Sparks E, Cirino AL, Depalma S, Colan SD, Funke B, Zimmerman RS, Robinson P, Watkins H, Seidman CE, Seidman JG, McNally EM, Ho C (2010) Familial dilated cardiomyopathy caused by an alpha-tropomyosin mutation. J Am Coll Cardiol 55:320–329CrossRefPubMedGoogle Scholar
  11. Lehrer SS, Golistina NL, Geeves MA (1997) Actin-tropomyosin activation of myosin subfragment 1 ATPase and thin filament cooperativity. The role of tropomyosin flexibility and end-to-end interactions. Biochemistry 36(44):13449–13454CrossRefPubMedGoogle Scholar
  12. Millar NC, Homsher E (1990) The effect of phosphate and calcium on force generation in glycerinated rabbit skeletal muscle fibers. A steady-state and transient kinetic study. J Biol Chem 265:20234–20240PubMedGoogle Scholar
  13. Monteiro PB, Lataro RC, Ferro JA, de Reinach FC (1994) Functional alpha-tropomyosin produced in Escherichia coli A dipeptide extension can substitute the amino-terminal acetyl group. J Biol Chem 269(14):10461–10466PubMedGoogle Scholar
  14. Muthuchamy M, Grupp IL, Grupp G, O’Toole BA, Kier AB, Boivin GP, Neumann J, Wieczorek DF (1995) Molecular and physiological effects of overexpressing striated muscle beta-tropomyosin in the adult murine heart. J Biol Chem 270(51):30593–30603CrossRefPubMedGoogle Scholar
  15. Olson TM, Kishimoto NY, Whitby FG, Michels VV (2001) Mutations that alter the surface charge of alpha-tropomyosin are associated with dilated cardiomyopathy. J Mol Cell Cardiol 33:723–732CrossRefPubMedGoogle Scholar
  16. Patton C, Thompson S, Epel D (2004) Some precautions in using chelators to buffer metals in biological solutions. Cell Calcium 35:427–431CrossRefPubMedGoogle Scholar
  17. Pieples K, Arteaga G, Solaro RJ, Grupp I, Lorenz JN, Boivin GP, Jagatheesan G, Labitzke E, de Tombe PP, Konhilas JP, Irving TC, Wieczorek DF (2002) Tropomyosin 3 expression leads to hypercontractility and attenuates myofilament length-dependent Ca2+ activation. Am J Physiol Heart Circ Physiol 283:H1344–H1353PubMedGoogle Scholar
  18. Rajan S, Ahmed RPH, Jagatheesan G, Petrashevskaya N, Boivin GP, Urboniene D, Arteaga GM, Wolska BM, Solaro RJ, Liggett SB, Wieczorek DF (2007) Dilated cardiomyopathy mutant tropomyosin mice develop cardiac dysfunction with significantly decreased fractional shortening and myofilament calcium sensitivity. Circ Res 101(2):205–214. Erratum in: Circ Res. 2007 Sep 14;101(6):e80, 2007Google Scholar
  19. Rajan S, Jagatheesan G, Karam CN, Alves ML, Bodi I, Schwartz A, Bulcao CF, D’Souza KM, Akhter SA, Boivin GP, Dube DK, Petrashevskaya N, Herr AB, Hullin R, Liggett SB, Wolska BM, Solaro RJ, Wieczorek DF (2010) Molecular and functional characterization of a novel cardiac-specific human tropomyosin isoform. Circulation 121:410–418CrossRefPubMedGoogle Scholar
  20. Rao VS, Marongelli EN, Guilford WH (2009) Phosphorylation of tropomyosin extends cooperative binding of myosin beyond a single regulatory unit. Cell Motil Cytosckelet 66(1):10–23CrossRefGoogle Scholar
  21. Robinson P, Griffiths PJ, Watkins H, Redwood CS (2007) Dilated and hyperthrophic cardiomyopathy mutations in troponin and α-tropomyosin have opposing effects on the calcium affinity of cardiac thin filaments. Circ Res 101:1266–1273CrossRefPubMedGoogle Scholar
  22. Solaro RJ (2009) CK-1827452, a sarcomere-directed cardiac myosin activator for acute and chronic heart disease. IDrugs 12:243–251PubMedGoogle Scholar
  23. Solaro RJ, Lee JA, Kentich JC, Allen DG (1988) Effects of acidosis on ventricular muscle from adult and neonatal rats. Circ Res 63(4):779–787PubMedGoogle Scholar
  24. Sumandea MP, Pyle WG, Kobayashi T, de Tombe PP, Solaro RJ (2003) Identification of a functionally critical protein kinase C phosphorylation residue of cardiac troponin T. J Biol Chem 278:35135–35144CrossRefPubMedGoogle Scholar
  25. Sun YB, Irving M (2010) The molecular basis of the steep force-calcium relation in heart muscle. J Mol Cell Cardiol 48:859–865CrossRefPubMedGoogle Scholar
  26. Trybus KM, Taylor EW (1980) Kinetic studies of the cooperative binding of subfragment 1 to regulated actin. Proc Natl Acad Sci USA 77:7209–7213CrossRefPubMedGoogle Scholar
  27. Vahebi S, Ota A, Li M, Warren CM, de Tombe PP, Wang Y, Solaro RJ (2007) p38-MAPK induced dephosphorylation of alpha-tropomyosin is associated with depression of myocardial sarcomeric tension and ATPase activity. Circ Res 100(3):408–415CrossRefPubMedGoogle Scholar
  28. VanBuren P, Palmer BM (2010) Cooperative activation of the cardiac myofilament: the pivotal role of tropomyosin. Circulation 121:351–353CrossRefPubMedGoogle Scholar
  29. Wannenburg T, Janssen PM, de Tombe PP (1997) The Frank-Starling mechanism is not mediated by changes in rate of cross-bridge detachment. Am J Physiol 273(5 Pt 2):H2428–H2435PubMedGoogle Scholar
  30. Warren CM, Arteaga GM, Rajan S, Ahmed RPH, Wieczorek DF, Solaro RJ (2008) Use of 2-D DIGE analysis reveals altered phosphorylation in a tropomyosin mutant (Glu54Lys) linked to dilated cardiomyopathy. Proteomics 8:100–106CrossRefPubMedGoogle Scholar
  31. Wolska BM, Keller RS, Evans CC, Palmiter KA, Phillips RM, Muthuchamy M, Oehlenschlager J, Wieczorek DF, de Tombe PP, Solaro RJ (1999) Correlation between myofilaments response to Ca2+ and altered dynamics of contraction and relaxation in transgenic cardiac cells than express beta-tropomyosin. Circ Res 84:745–751PubMedGoogle Scholar
  32. Yuan C, Sheng Q, Tang H, Li Y, Zeng R, Solaro RJ (2008) Quantitative comparison of sarcomeric phosphoproteomes of neonatal and adult rat hearts. Am J Physiol Heart Circ Physiol 295:H647–H656CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Chehade N. Karam
    • 1
  • Chad M. Warren
    • 1
  • Sudarsan Rajan
    • 2
  • Pieter P. de Tombe
    • 3
  • David F. Wieczorek
    • 2
  • R. John Solaro
    • 1
  1. 1.Department of Physiology and Biophysics, Center for Cardiovascular Research, College of MedicineUniversity of Illinois at ChicagoChicagoUSA
  2. 2.Department of Molecular Genetics, Biochemistry and Microbiology, College of MedicineUniversity of CincinnatiCincinnatiUSA
  3. 3.Department of Cell and Molecular Physiology, Stritch School of MedicineLoyola University ChicagoChicagoUSA

Personalised recommendations