Abstract
Titin (also known as connectin) is a giant elastic protein located in the striated-muscle sarcomere where it spans from Z-line to M-line. A large part of the I-band region of the titin molecule is extensible and functions as a molecular spring that underlies passive muscle stiffness when sarcomeres are stretched. This spring has a complex composition. In cardiac titin it consists of three extensible elements: tandem Ig segments, the PEVK segment and the N2B unique sequence. Here we discuss our recent work focused on understanding the molecular basis of titin’s extensibility and in which force-extension curves were measured by using an atomic force microscope specialized for stretching single molecules. We will discuss results from recombinant proteins that represent the various elements of titin’s extensible region. The obtained single molecule mechanical characteristics of titin’s various spring elements explain well their measured extension in the cardiac sarcomere when stretched within their physiological length range. We also examined how titin’s contribution to passive muscle stiffness may be adjusted. We discuss evidence that suggests that calcium/S100 may adjust titin-based stiffness and that phosphorylation of cardiac titin’s N2B spring elements reduces titin-based passive stiffness in cardiac muscle. Finally, we show that the cardiac sarcomere of large mammals co-expresses titin isoforms and that differential splicing of titin’s spring elements is a long-term mechanism of adjustment, which plays a role in passive stiffness modulation during heart disease.
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References
C.C. Gregorio, H. Granzier, H. Sorimachi, and S. Labeit, Muscle assembly: a titanic achievement?, Curr Opin Cell Biol 11, 18–25. (1999).
L. Tskhovrebova, and J. Trinick, Role of titin in vertebrate striated muscle, Philos Trans R Soc Lond B Biol Sci 357, 199–206 (2002).
K. Wang, Cytoskeletal matrix in striated muscle: the role of titin, nebulin and intermediate filaments, Adv Exp Med Biol 170, 285–305 (1984).
K. Maruyama, Connectin/titin, giant elastic protein of muscle, FASEB J 11, 341–345. (1997).
W. A. Linke, Stretching molecular springs: elasticity of titin filaments in vertebrate striated muscle, Histol Histopathol 15, 799–811 (2000).
H. Granzier, and S. Labeit, Cardiac titin: an adjustable multi-function spring, Journal of Physiology (London) 541, 335–342 (2002).
K. Wang, Titin/connectin and nebulin: giant protein rulers of muscle structure and function, Adv Biophys 33, 123–134 (1996).
S. Improta, A. S. Politou, and A. Pastore, Immunoglobulin-like modules from titin I-band: extensible components of muscle elasticity, Structure 4, 323–337. (1996).
S. Labeit, and B. Kolmerer, Titins: giant proteins in charge of muscle ultrastructure and elasticity, Science 270, 293–296 (1995).
A. Freiburg, K. Trombitas, W. Hell, O. Cazorla, F. Fougerousse, T. Centner, B. Kolmerer, C. Witt, J. S. Beckmann, C. C. Gregorio, H. Granzier, and S. Labeit, Series of exon-skipping events in the elastic spring region of titin as the structural basis for myofibrillar elastic diversity, Circ Res 86, 1114–1121. (2000).
K. Trombitas, M Greaser, S. Labeit, J. P. Jin, M Kellermayer, M. Helmes, and H. Granzier, Titin extensibility in situ: entropic elasticity of permanently folded and permanently unfolded molecular segments, J Cell Biol 140, 853–859 (1998).
W. A Linke, and H. Granzier, A spring tale: new facts on titin elasticity, Biophys J 75, 2613–2614. (1998).
W. A Linke, M. R. Stockmeier, M. Ivemeyer, H. Hosser, and P. Mündel, Characterizing titin’s I-band 1g domain region as an entropic spring, J Cell Sci 111( Pt 11), 1567–1574 (1998).
W. A Linke, M. Ivemeyer, P. Mündel, M. R. Stockmeier, and B. Kolmerer, Nature of PEVK-titin elasticity in skeletal muscle, Proc Natl Acad Sci U S A 95, 8052–8057 (1998).
L. Tskhovrebova, and J. Trinick, Flexibility and extensibility in the titin molecule: analysis of electron microscope data, J Mol Biol 310, 755–771. (2001).
A. Minajeva, M. Kulke, J. M. Fernandez, and W. A. Linke, Unfolding of titin domains explains the viscoelastic behavior of skeletal myofibrils, Biophys J 80, 1442–1451. (2001).
K. Trombitas, M. Greaser, S. Labeit, J. P. Jin, M. Kellermayer, M. Helmes, and H. Granzier, Titin extensibility in situ: entropie elasticity of permanently folded and permanently unfolded molecular segments, J Cell Biol 140, 853–859. (1998).
K. Trombitas, M. Greaser, G. French, and H. Granzier, PEVK extension of human soleus muscle titin revealed by immunplabeling with the anti-titin antibody 9D10, J Struct Biol 122, 188–196 (1998).
K. Ma, L. Kan, and K. Wang, Polyproline II helix is a key structural motif of the elastic PEVK segment of titin, Biochemistry 40, 3427–3438. (2001).
H. Li, A. F. Oberhäuser, S. D. Redick, M. Carrion-Vazquez, H. P. Erickson, and J. M. Fernandez, Multiple conformations of PEVK proteins detected by single-molecule techniques, Proc Natl Acad Sci U S A 98, 10682–10686.(2001).
J. F. Marko, and E. D. Siggia, Fluctuations and supercoiling of DNA, Science 265, 506–508 (1994).
C. Bustamante, J. F. Marko, E. D. Siggia, and S. Smith, Entropie elasticity of lambda-phage DNA, Science 265, 1599–1600 (1994).
M. Helmes, K. Trombitas, T. Centner, M. Kellermayer, S. Labeit, W. A. Linke, and H. Granzier, Mechanically driven contour-length adjustment in rat cardiac titin’s unique N2B sequence: titin is an adjustable spring, Circ Res 84, 1339–1352. (1999).
K. Trombitas, A. Freiburg, T. Centner, S. Labeit, and H. Granzier, Molecular dissection of N2B cardiac titin’s extensibility, Biophys J 77, 3189–3196. (1999).
K. Watanabe, P. Nair, D. Labeit, M. S. Kellermayer, M. Greaser, S. Labeit, and H. Granzier, Molecular mechanics of cardiac titin’s PEVK and N2B spring elements, J Biol Chem 277, 11549–11558 (2002).
M. Carrion-Vazquez, P. E. Marszalek, A. F. Oberhauser, and J. M. Fernandez, Atomic force microscopy captures length phenotypes in single proteins, Proc Natl Acad Sci U S A 96, 11288–11292. (1999).
M. Carrion-Vazquez, A. F. Oberhauser, S. B. Fowler, P. E. Marszalek, S. E. Broedel, J. Clarke, and J. M. Fernandez, Mechanical and chemical unfolding of a single protein: a comparison, Proc Natl Acad Sci U S A 96, 3694–3699. (1999).
M. Rief, M. Gautel, F. Oesterhelt, J. M. Fernandez, and H. E. Gaub, Reversible unfolding of individual titin immunoglobulin domains by AFM, Science 276, 1109–1112. (1997).
P. E. Marszalek, H. Lu, H. Li, M. Carrion-Vazquez, A. F. Oberhauser, K. Schulten, and J. M. Fernandez, Mechanical unfolding intermediates in titin modules, Nature 402, 100–103. (1999).
K. Watanabe, C. Muhle-Goll, M. S. Kellermayer, S. Labeit, and H. Granzier, Different molecular mechanics displayed by titin’s constitutively and differentially expressed tandem Ig segments, J Struct Biol 137, 248–258 (2002).
M. Carrion-Vazquez, A. F. Oberhauser, T. E. Fisher, P. E. Marszalek, H. Li, and J. M. Fernandez, Mechanical design of proteins studied by single-molecule force spectroscopy and protein engineering, Prog Biophys Mol Biol 74, 63–91 (2000).
R. Yamasaki, M. Bern, Y. Wu, K. Trombitas, M. McNabb, M. S. Kellermayer, C. Witt, D. Labeit, S. Labeit, M. Greaser, and H. Granzier, Titin-actin interaction in mouse myocardium: passive tension modulation and its regulation by calcium/S100Al, Biophys J 81, 2297–2313 (2001).
M. Kulke, S. Fujita-Becker, E. Rostkova, C. Neagoe, D. Labeit, D. J. Manstein, M. Gautel, and W. A. Linke, Interaction between PEVK-titin and actin filaments: origin of a viscous force component in cardiac myofibrils, Circ Res 89, 874–881 (2001).
W. Kabsch, H. G. Mannherz, D. Suck, E. F. Pai, and K. C. Holmes, Atomic structure of the actin:DNase I complex, Nature 347, 37–44. (1990).
G. Fulgenzi, L. Graciotti, A. L Granata, A. Corsi, P. Fucini, A. A. Noegel, H. M. Kent, and M. Stewart, Location of the binding site of the mannose-specific lectin comitin on F-actin, J Mol Biol 284, 1255–1263. (1998).
E. Friederich, K. Vancompemolle, C. Huet, M. Goethals, J. Finidori, J. Vandekerckhove, and D. Louvard, An actin-binding site containing a conserved motif of charged amino acid residues is essential for the morphogenic effect of villin, Cell 70, 81–92. (1992).
M. Pfuhl, S. J. Winder, and A Pastore, Nebulin, a helical actin binding protein, Embo J 13, 1782–1789. (1994).
R. Yamasaki, Y. Wu, M. McNabb, M. Greaser, S. Labeit, and H. Granzier, Protein kinase A phosphorylates titin’s cardiac-specific N2B domain and reduces passive tension in rat cardiac myocytes, Circ Res 90, 1181–1188 (2002).
K. T. Strang, N. K. Sweitzer, M. L. Greaser, and R. L. Moss, Beta-adrenergic receptor stimulation increases unloaded shortening velocity of skinned single ventricular myocytes from rats, Circ Res 74, 542–549. (1994).
R. J. Solaro, A. J. Moir, and S. V. Perry, Phosphorylation of troponin I and the inotropic effect of adrenaline in the perfused rabbit heart, Nature 262, 615–617 (1976).
H. C. Hartzell, and D. B. Glass, Phosphorylation of purified cardiac muscle C-protein by purified cAMP-dependent and endogenous Ca2+-calmodulin-dependent protein kinases, J Biol Chem 259, 15587–15596 (1984).
K. T. Strang, and R. L. Moss, Alpha 1-adrenergic receptor stimulation decreases maximum shortening velocity of skinned single ventricular myocytes from rats, Circ Res 77, 114–120 (1995).
M. L. Bang, T. Centner, F. Fornoff, A J. Geach, M. Gotthardt, M. McNabb, C. C. Witt, D. Labeit, C. C. Gregorio, H. Granzier, and S. Labeit, The complete gene sequence of titin, expression of an unusual approximately 700-kDa titin isoform, and its interaction with obscurin identify a novel Z-line to I-band linking system, Circ Res 89, 1065–1072 (2001).
M. Greaser, Identification of new repeating motifs in titin, Proteins 43, 145–149. (2001).
O. Cazorla, A. Freiburg, M. Helmes, T. Centner, M. McNabb, Y. Wu, K. Trombitas, S. Labeit, and H. Granzier, Differential expression of cardiac titin isoforms and modulation of cellular stiffness, Circ Res 86, 59–67. (2000).
R. Horowits, and R. J. Podolsky, The positional stability of thick filaments in activated skeletal muscle depends on sarcomere length: evidence for the role of titin filaments, J Cell Biol 105, 2217–2223 (1987).
M. Helmes, K. Trombitas, and H. Granzier, Titin develops restoring force in rat cardiac myocytes, Circ Res 79, 619–626 (1996).
H. Granzier, M. Kellermayer, M. Helmes, and K. Trombitas, Titin elasticity and mechanism of passive force development in rat cardiac myocytes probed by thin-filament extraction, Biophys J 73, 2043–2053. (1997).
H. Granzier, M. Helmes, O. Cazorla, M. McNabb, D. Labeit, Y. Wu, R. Yamasaki, A. Redkar, M. Kellermayer, S. Labeit, and K. Trombitas, Mechanical properties of titin isoforms, Adv Exp Med Biol 481, 283–300 (2000).
W. A Linke, M. Ivemeyer, S. Labeit, H. Hinssen, J. C. Ruegg, and M. Gautel, Actin-titin interaction in cardiac myofibrils: probing a physiological role, Biophys J 73, 905–919. (1997).
K. Trombitas, Y. Wu, D. Labeit, S. Labeit, and H. Granzier, Cardiac titin isoforms are coexpressed in the half-sarcomere and extend independently, Am J Physiol Heart Circ Physiol 281, H1793–1799 (2001).
S. Labeit, and B. Kolmerer, Titins: giant proteins in charge of muscle ultrastructure and elasticity, Science 270, 293–296. (1995).
S. Hein, and J. Schaper, Pathogenesis of dilated cardiomyopathy and heart failure: insights from cell morphology and biology, Curr Opin Cardiol 11, 293–301. (1996).
S. Hein, D. Scholz, N. Fujitani, H. Rennollet, T. Brand, A. Friedl, and J. Schaper, Altered expression of titin and contractile proteins in failing human myocardium, J Mol Cell Cardiol 26, 1291–1306. (1994).
I. Morano, K. Hadicke, S. Grom, A. Koch, R. H. Schwinger, M. Bohm, S. Bartel, E. Erdmann, and E. G. Krause, Titin, myosin light chains and C-protein in the developing and failing human heart, J Mol Cell Cardiol 26, 361–368. (1994).
B. L. Siu, H. Niimura, J. A. Osbome, D. Fatkin, C. MacRae, S. Solomon, D. W. Benson, J. G. Seidman, and C. E. Seidman, Familial dilated cardiomyopathy locus maps to chromosome 2q31, Circulation 99, 1022–1026. (1999).
M. Itoh-Satoh, T. Hayashi, H. Nishi, Y. Koga, T. Arimura, T. Koyanagi, M. Takahashi, S. Hohda, K. Ueda, T. Nouchi, M. Hiroe, F. Marumo, T. Imaizumi, M Yasunami, and A. Kimura, Titin Mutations as the Molecular Basis for Dilated Cardiomyopathy, Biochem Biophys Res Commun 291, 385–393 (2002).
B. Gerull, M. Gramlich, J. Atherton, M. McNabb, K. Trombitas, S. Sasse-Klaassen, J. G. Seidman, C. Seidman, H. Granzier, S. Labeit, M. Frenneaux, and L Thierfelder, Mutations of TTN, encoding the giant muscle filament titin, cause familial dilated cardiomyopathy, Nat Genet 30, 201–204. (2002).
X. Xu, S. E. Meiler, T. P. Zhong, M. Mohideen, D. A Crossley, W. W. Burggren, and M. C Fishman, Cardiomyopathy in zebrafish due to mutation in an alternatively spliced exon of titin, Nat Genet 30, 205–209 (2002).
P. Hackman, A. Vihola, H. Haravuori, S. Marchand, J. Sarparanta, J. De Seze, S. Labeit, C. Witt, L Peltonen, I. Richard, and B. Udd, Tibial Muscular Dystrophy Is a Titinopathy Caused by Mutations in TTN, the Gene Encoding the Giant Skeletal-Muscle Protein Titin, Am J Hum Genet 71, 492–500 (2002).
T. Centner, J. Yano, E. Kimura, A. S. McElhinny, K. Pelin, C. C. Witt, M. L. Bang, K. Trombitas, H. Granzier, C. C. Gregorio, H. Sorimachi, and S. Labeit, Identification of muscle specific ring finger proteins as potential regulators of the titin kinase domain, J Mol Biol 306, 717–726 (2001).
M. Gotthardt, R. E. Hammer, N. Hubner, J. Monti, C. C. Witt, M. McNabb, J. A. Richardson, H. Granzier, S. Labeit, and J. Herz, Conditional expression of mutant M-line titins results in cardiomyopathy with altered sarcomere structure, J Biol Chem (2002).
Y. Wu, S. P. Bell, K. Trombitas, C. C. Witt, S. Labeit, M. M. Le Winter, and H. Granzier, Changes in titin isoform expression in pacing-induced cardiac failure give rise to increased passive muscle stiffness, Circulation 106, 1384–1389 (2002).
Y. Wu, S. Bell, M. M. Le Winter, S. Labeit, and H. Granzier, Titin: an endosarcomeric protein that modulates myocardial stiffness., Journal of Cardiac Failure 8, S276–S286 (2002).
C. Neagoe, M. Kulke, F. del Monte, J. K. Gwathmey, P. P. de Tombe, R. J. Hajjar, and W. A. Linke, Titin isoform switch in ischémie human heart disease, Circulation 106, 1333–1341 (2002).
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Granzier, H. et al. (2003). Adaptations in Titin’s Spring Elements in Normal and Cardiomyopathic Hearts. In: Sugi, H. (eds) Molecular and Cellular Aspects of Muscle Contraction. Advances in Experimental Medicine and Biology, vol 538. Springer, Boston, MA. https://doi.org/10.1007/978-1-4419-9029-7_46
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DOI: https://doi.org/10.1007/978-1-4419-9029-7_46
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