Abstract
The cellular function of the giant protein titin in striated muscle is a major focus of scientific attention. Particularly, its role in passive mechanics has been extensively investigated. In strong contrast, the structural details of this filament are very poorly understood. To date, only a handful of atomic models from single domain components have become available and data on poly-constructs are limited to scarce SAXS analyses. In this study, we examine the molecular parameters of poly-Ig tandems from I-band titin relevant to muscle elasticity. We revisit conservation patterns in domain and linker sequences of I-band modules and interpret these in the light of available atomic structures of Ig domains from muscle proteins. The emphasis is placed on features expected to affect inter-domain arrangements. We examine the overall conformation of a 6Ig fragment, I65–I70, from the skeletal I-band of soleus titin using SAXS and electron microscopy approaches. The possible effect of highly conserved glutamate groups at the linkers as well as the ionic strength of the medium on the overall molecular parameters of this sample is investigated. Our findings indicate that poly-Ig tandems from I-band titin tend to adopt extended arrangements with low or moderate intrinsic flexibility, independently of the specific features of linkers or component Ig domains across constitutively- and differentially-expressed tandems. Linkers do not appear to operate as free hinges so that lateral association of Ig domains must occur infrequently in samples in solution, even that inter-domain sequences of 4–5 residues length would well accommodate such geometry. It can be expected that this principle is generally applicable to all Ig-tandems from I-band titin.
Similar content being viewed by others
References
Atkinson RA, Joseph C, Kelly G, Muskett FW, Frenkiel TA, Nietlispach D, Pastore A, (2001). Ca2+-independent binding of an EF-hand domain to a novel motif in the alpha-actinin-titin complexNat Struct Biol 8: 853–857
Bang ML, Centner T, Fornoff F, Geach AJ, Gotthardt M, McNabb M, Witt CC, Labeit D, Gregorio CC, Granzier H, Labeit S, (2001). 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 systemCirc Res 89: 1065–1072
Bullard B, Linke WA, Leonard K, (2002). Varieties of elastic protein in invertebrate musclesJ Muscle Res Cell Motil 23: 435–447
Di Cola E, Waigh T, Trinick J, Tskhovrebova L, Houmeida A, Pyckhout-Hintzen W, Dewhurst, (2005). Persistence Length of Titin from Rabbit Skeletal Muscles Measured with Scattering and Microrheology TechniquesBiophys J. 88: 4095–4106
Feigin LA, Svergun DI, (1987). Structure Analysis by Small-Angle X-ray and Neutron Scattering, Plenum Press, New York
Flory PJ, (1969). Statistical Mechanics of Chain Molecules, Interscience publishers, New York
Fong S, Hamill SJ, Proctor M, Freund SM, Benian GM, Chothia C, Bycroft M, Clarke J, (1996). Structure and stability of an immunoglobulin superfamily domain from twitchin, a muscle protein of the nematode Caenorhabditis elegansJ Mol Biol 264: 624–639
Gao M, Lu H, Schulten K, (2001). Simulated refolding of stretched titin immunoglobulin domainsBiophys J 81: 2268–2277
Gao M, Wilmanns M, Schulten K, (2002). Steered molecular dynamics studies of titin I1 domain unfoldingBiophys J 83: 3435–3445
Gautel M, (1996). The super-repeats of titin/connectin and their interactions: glimpses at sarcomeric assemblyAdv Biophys 33: 27–37
Gautel M, Goulding D, (1996). A molecular map of titin/connectin elasticity reveals two different mechanisms acting in seriesFEBS Lett 385: 11–14
Granzier HL, Labeit S, (2004). The giant protein titin: a major player in myocardial mechanics, signaling, and diseaseCirc Res 94: 284–295
Higuchi H, Nakauchi Y, Maruyama K, Fujime S, (1993). Characterization of beta-connectin (titin 2) from striated muscle by dynamic light scatteringBiophys J. 65: 1906–1915
Holden HM, Ito M, Hartshorne DJ, Rayment I, (1992). X-ray structure determination of telokin, the C-terminal domain of myosin light chain kinase, at 2.8 A resolutionJ Mol Biol 227: 840–851
Improta S, Krueger JK, Gautel M, Atkinson RA, Lefevre JF, Moulton S, Trewhella J, Pastore A, (1998). The assembly of immunoglobulin-like modules in titin: implications for muscle elasticityJ Mol Biol 284: 761–777
Improta S, Politou AS, Pastore A, (1996). Immunoglobulin-like modules from titin I-band: extensible components of muscle elasticityStructure 4: 323–337
Kellermayer MS, Bustamante C, Granzier HL, (2003). Mechanics and structure of titin oligomers explored with atomic force microscopyBiochim Biophys Acta 1604: 105–114
Kellermayer MS, Smith SB, Granzier HL, Bustamante C, (1997). Folding-unfolding transitions in single titin molecules characterized with laser tweezersScience 276: 1112–1116
Kenny PA, Liston EM, Higgins DG, (1999). Molecular evolution of immunoglobulin and fibronectin domains in titin and related muscle proteinsGene 17: 11–23
Kobe B, Heierhorst J, Feil SC, Parker MW, Benian GM, Weiss KR, Kemp BE, (1996). Giant protein kinases: domain interactions and structural basis of autoregulationEMBO J 15: 6810–6821
Konarev PV, Volkov VV, Sokolova AV, Koch MHJ, Svergun DI, (2003). PRIMUS – a Windows-PC based system for small-angle scattering data analysisJ Appl Crystallogr 36: 1277–1282
Labeit S, Kolmerer B, (1995). Titins: giant proteins in charge of muscle ultrastructure and elasticityScience 270: 293–296
Li H, Carrion-Vazquez M, Oberhauser AF, Marszalek PE, Fernandez JM, (2000). Point mutations alter the mechanical stability of immunoglobulin modulesNat Struct Biol 7: 1117–1120
Li H, Fernandez JM, (2003). Mechanical design of the first proximal Ig domain of human cardiac titin revealed by single molecule force spectroscopy. J Mol Biol 334: 75–86
Li H, Linke WA, Oberhauser AF, Carrion-Vazquez M, Kerkvliet JG, Lu H, Marszalek PE, Fernandez JM, (2002). Reverse engineering of the giant muscle protein titinNature 418: 998–1002
Linke WA, Ivemeyer M, Olivieri N, Kolmerer B, Ruegg JC, Labeit S, (1996). Towards a molecular understanding of the elasticity of titinJ Mol Biol 261: 62–71
Lu H, Isralewitz B, Krammer A, Vogel V, Schulten K, (1998). Unfolding of titin immunoglobulin domains by steered molecular dynamics simulationBiophys J 75: 662–671
Ma K, Kan L, Wang K, (2001). Polyproline II helix is a key structural motif of the elastic PEVK segment of titinBiochemistry 40: 3427–3438
Ma K, Wang K, (2003). Malleable conformation of the elastic PEVK segment of titin: non-co-operative interconversion of polyproline II helix, beta-turn and unordered structuresBiochem J 374: 687–695
Maruyama K, Matsubara S, Natori R, Nonomura Y, Kimura S, Ohashi K, murakami F, Handa S, Eguchi G, (1977). Connectin, an elastic protein of muscle: characterization and functionJ Biochem 82: 317–337
Mayans O, van der Ven PF, Wilm M, Mues A, Young P, Furst DO, Wilmanns M, Gautel M, (1998). Structural basis for activation of the titin kinase domain during myofibrillogenesisNature 395: 863–869
Mayans O, Wuerges J, Canela S, Gautel M, Wilmanns M, (2001). Structural evidence for a possible role of reversible disulphide bridge formation in the elasticity of the muscle protein titinStructure 9: 331–340
Miller MK, Granzier H, Ehler E, Gregorio CC, (2004). The sensitive giant: the role of titin-based stretch sensing complexes in the heartTrends Cell Biol 14: 119–126
Muhle-Goll CM, Pastore A, Nilges M, (1998). The three-dimensional structure of a type I module from titin: a prototype of intracellular fibronectin type III domainsStructure 6: 1291–1302
Pfuhl M, Pastore A, (1995). Tertiary structure of an immunoglobulin-like domain from the giant muscle protein titin: a new member of the I setStructure 3: 391–401
Politou AS, Gautel M, Improta S, Vangelista L, Pastore A, (1996). The elastic I-band region of titin is assembled in a “modular” fashion by weakly interacting Ig-like domainsJ Mol Biol 255: 604–616
Rief M, Gautel M, Oesterhelt F, Fernandez JM, Gaub HE, (1997). Reversible unfolding of individual titin immunoglobulin domains by AFMScience 276: 1109–1112
Rivetti C, Guthold M, Bustamante C, (1996). Scanning force microscopy of DNA deposited onto mica: equilibration versus kinetic trapping studied by statistical polymer chain analysisJ Mol Biol. 264: 919–932
Scott KA, Steward A, Fowler SB, Clarke J, (2002). Titin; a multidomain protein that behaves as the sum of its partsJ Mol Biol 315: 819–829
Svergun DI, (1992). Determination of the regularization parameter in indirect transform methods using perceptual criteriaJ Appl Crystallogr 25: 495–503
Svergun DI, (1993). A direct indirect method of small-angle scattering data treatmentJ Appl Crystallogr 26: 258–267
Trombitas K, Greaser M, Labeit S, Jin JP, Kellermayer M, Helmes M, Granzier H, (1998). Titin extensibility in situ: entropic elasticity of permanently folded and permanently unfolded molecular segmentsJ Cell Biol 140: 853–859
Tskhovrebova L, Trinick J, (2001). Flexibility and extensibility in the titin molecule: analysis of electron microscope dataJ Mol Biol 310: 755–771
Tskhovrebova L, Trinick J, (2003). Titin: properties and family relationshipsNat Rev Mol Cell Biol 4: 679–689
Tskhovrebova L, Trinick J, Sleep JA, Simmons RM, (1997). Elasticity and unfolding of single molecules of the giant muscle protein titinNature 387: 308–312
Watanabe K, Muhle-Goll C, Kellermayer MS, Labeit S, Granzier H, (2002). Different molecular mechanics displayed by titin’s constitutively and differentially expressed tandem Ig segmentsJ Struct Biol 137: 248–258
Witt CC, Olivieri N, Centner T, Kolmerer B, Millevoi S, Morell J, Labeit D, Labeit S, Jockusch H, Pastore A, (1998). A survey of the primary structure and the interspecies conservation of I-band titin’s elastic elements in vertebratesJ Struct Biol 122: 206–215
Zou P, Gautel M, Geerlof A, Wilmanns M, Koch MH, Svergun DI, (2003). Solution scattering suggests cross-linking function of telethonin in the complex with titinJ Biol Chem 278: 2636–2644
Acknowledgements
Our gratitude goes to Prof. Ueli Aebi for support with the electron microscopy technique. We would like to acknowledge financial support to D.L. by D.F.G. (La1969/1-1) and to L.K. by the Swiss Society for Research on Muscular Diseases (grant awarded to U. Aebi and S. Strelkov).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Marino, M., Svergun, D.I., Kreplak, L. et al. Poly-Ig tandems from I-band titin share extended domain arrangements irrespective of the distinct features of their modular constituents. J Muscle Res Cell Motil 26, 355–365 (2005). https://doi.org/10.1007/s10974-005-9017-6
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10974-005-9017-6