{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,27]],"date-time":"2026-04-27T20:43:23Z","timestamp":1777322603201,"version":"3.51.4"},"reference-count":41,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2019,3,26]],"date-time":"2019-03-26T00:00:00Z","timestamp":1553558400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2019,3,26]],"date-time":"2019-03-26T00:00:00Z","timestamp":1553558400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Patients with Duchenne muscular dystrophy (DMD) lack the protein dystrophin, which is a critical molecular component of the dystrophin-glycoprotein complex (DGC). Dystrophin is hypothesized to function as a molecular shock absorber that mechanically stabilizes the sarcolemma of striated muscle through interaction with the cortical actin cytoskeleton via its N-terminal half and with the transmembrane protein \u03b2-dystroglycan via its C-terminal region. Utrophin is a fetal homologue of dystrophin that can subserve many dystrophin functions and is therefore under active investigation as a dystrophin replacement therapy for DMD. Here, we report the first mechanical characterization of utrophin using atomic force microscopy (AFM). Our data indicate that the mechanical properties of spectrin-like repeats in utrophin are more in line with the PEVK and Ig-like repeats of titin rather than those reported for repeats in spectrin or dystrophin. Moreover, we measured markedly different unfolding characteristics for spectrin repeats within the N-terminal actin-binding half of utrophin compared to those in the C-terminal dystroglycan-binding half, even though they exhibit identical thermal denaturation profiles. Our results demonstrate dramatic differences in the mechanical properties of structurally homologous utrophin constructs and suggest that utrophin may function as a stiff elastic element in series with titin at the myotendinous junction.<\/jats:p>","DOI":"10.1038\/s41598-019-41569-4","type":"journal-article","created":{"date-parts":[[2019,3,26]],"date-time":"2019-03-26T11:03:26Z","timestamp":1553598206000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Distinct mechanical properties in homologous spectrin-like repeats of utrophin"],"prefix":"10.1038","volume":"9","author":[{"given":"Sivaraman","family":"Rajaganapathy","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jackie L.","family":"McCourt","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sayan","family":"Ghosal","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angus","family":"Lindsay","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Preston M.","family":"McCourt","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dawn A.","family":"Lowe","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"James M.","family":"Ervasti","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Murti V.","family":"Salapaka","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2019,3,26]]},"reference":[{"key":"41569_CR1","doi-asserted-by":"publisher","first-page":"304","DOI":"10.1002\/ana.23528","volume":"71","author":"JR Mendell","year":"2012","unstructured":"Mendell, J. R. et al. Evidence-based path to newborn screening for Duchenne muscular dystrophy. Ann. Neurol. 71, 304\u201313 (2012).","journal-title":"Ann. Neurol."},{"key":"41569_CR2","doi-asserted-by":"publisher","first-page":"919","DOI":"10.1016\/0092-8674(87)90579-4","volume":"51","author":"EP Hoffman","year":"1987","unstructured":"Hoffman, E. P., Brown, R. H. & Kunkel, L. M. Dystrophin: The Protein Product of the Duchenne Muscular Dystrophy Locus. Cell 51, 919\u2013928 (1987).","journal-title":"Cell"},{"key":"41569_CR3","doi-asserted-by":"publisher","first-page":"13591","DOI":"10.1074\/jbc.R200021200","volume":"278","author":"JM Ervasti","year":"2003","unstructured":"Ervasti, J. M. Costameres: the Achilles\u2019 heel of Herculean muscle. J. Biol. Chem. 278, 13591\u20134 (2003).","journal-title":"J. Biol. Chem."},{"key":"41569_CR4","doi-asserted-by":"publisher","first-page":"1209","DOI":"10.1083\/jcb.150.5.1209","volume":"150","author":"IN Rybakova","year":"2000","unstructured":"Rybakova, I. N., Patel, J. R. & Ervasti, J. M. The Dystrophin Complex Forms a Mechanically Strong Link between the Sarcolemma and Costameric Actin. J. Cell Biol. 150, 1209\u20131214 (2000).","journal-title":"J. Cell Biol."},{"key":"41569_CR5","doi-asserted-by":"publisher","first-page":"108","DOI":"10.1016\/j.bbadis.2006.05.010","volume":"1772","author":"JM Ervasti","year":"2007","unstructured":"Ervasti, J. M. Dystrophin, its interactions with other proteins, and implications for muscular dystrophy. Biochim. Biophys. Acta 1772, 108\u201317 (2007).","journal-title":"Biochim. Biophys. Acta"},{"key":"41569_CR6","doi-asserted-by":"publisher","first-page":"1441","DOI":"10.1038\/4033","volume":"4","author":"J Tinsley","year":"1998","unstructured":"Tinsley, J. et al. Expression of full-length utrophin prevents muscular dystrophy in mdx mice. Nat. Med. 4, 1441\u20134 (1998).","journal-title":"Nat. Med."},{"key":"41569_CR7","doi-asserted-by":"publisher","first-page":"1458","DOI":"10.1113\/EP085308","volume":"100","author":"S Guiraud","year":"2015","unstructured":"Guiraud, S., Chen, H., Burns, D. T. & Davies, K. E. Advances in genetic therapeutic strategies for Duchenne muscular dystrophy. Exp. Physiol. 100, 1458\u20131467 (2015).","journal-title":"Exp. Physiol."},{"key":"41569_CR8","doi-asserted-by":"publisher","first-page":"585","DOI":"10.1016\/0736-5748(95)00039-J","volume":"13","author":"C Rigoletto","year":"1995","unstructured":"Rigoletto, C. et al. Utrophin expression during human fetal development. Int. J. Dev. Neurosci. 13, 585\u2013593 (1995).","journal-title":"Int. J. Dev. Neurosci."},{"key":"41569_CR9","doi-asserted-by":"publisher","first-page":"499","DOI":"10.1016\/0896-6273(91)90301-F","volume":"7","author":"K Ohlendieck","year":"1991","unstructured":"Ohlendieck, K. et al. Dystrophin-related protein is localized to neuromuscular junctions of adult skeletal muscle. Neuron 7, 499\u2013508 (1991).","journal-title":"Neuron"},{"key":"41569_CR10","doi-asserted-by":"publisher","first-page":"795","DOI":"10.1016\/j.jmb.2005.07.064","volume":"352","author":"N Bhasin","year":"2005","unstructured":"Bhasin, N. et al. Molecular extensibility of mini-dystrophins and a dystrophin rod construct. J. Mol. Biol. 352, 795\u2013806 (2005).","journal-title":"J. Mol. Biol."},{"key":"41569_CR11","doi-asserted-by":"publisher","first-page":"796","DOI":"10.1002\/cm.20489","volume":"67","author":"CC Krieger","year":"2010","unstructured":"Krieger, C. C. et al. Exon-skipped dystrophins for treatment of Duchenne muscular dystrophy: mass spectrometry mapping of most exons and cooperative domain designs based on single molecule mechanics. Cytoskelet. 67, 796\u2013807 (2010).","journal-title":"Cytoskelet."},{"key":"41569_CR12","doi-asserted-by":"publisher","first-page":"12140","DOI":"10.1021\/acsnano.8b05721","volume":"12","author":"S Le","year":"2018","unstructured":"Le, S. et al. Dystrophin As a Molecular Shock Absorber. ACS Nano 12, 12140\u201312148 (2018).","journal-title":"ACS Nano"},{"key":"41569_CR13","doi-asserted-by":"publisher","first-page":"1109","DOI":"10.1126\/science.276.5315.1109","volume":"276","author":"M Rief","year":"1997","unstructured":"Rief, M., Gautel, M., Oesterhelt, F., Fernandez, J. M. & Gaub, H. E. Reversible unfolding of individual titin immunoglobulin domains by AFM. Science 276, 1109\u20131112 (1997).","journal-title":"Science"},{"key":"41569_CR14","doi-asserted-by":"publisher","first-page":"3694","DOI":"10.1073\/pnas.96.7.3694","volume":"96","author":"M Carrion-Vazquez","year":"1999","unstructured":"Carrion-Vazquez, M. et al. Mechanical and chemical unfolding of a single protein: a comparison. Proc. Natl. Acad. Sci. USA 96, 3694\u20133699 (1999).","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"41569_CR15","doi-asserted-by":"publisher","first-page":"553","DOI":"10.1006\/jmbi.1998.2466","volume":"286","author":"M Rief","year":"1999","unstructured":"Rief, M., Pascual, J., Saraste, M. & Gaub, H. E. Single molecule force spectroscopy of spectrin repeats: low unfolding forces in helix bundles. J. Mol. Biol. 286, 553\u2013561 (1999).","journal-title":"J. Mol. Biol."},{"key":"41569_CR16","doi-asserted-by":"publisher","first-page":"3286","DOI":"10.1016\/S0006-3495(03)74747-X","volume":"85","author":"R Law","year":"2003","unstructured":"Law, R. et al. Pathway shifts and thermal softening in temperature-coupled forced unfolding of spectrin domains. Biophys. J. 85, 3286\u20133293 (2003).","journal-title":"Biophys. J."},{"key":"41569_CR17","doi-asserted-by":"publisher","first-page":"533","DOI":"10.1016\/S0006-3495(03)74872-3","volume":"84","author":"R Law","year":"2003","unstructured":"Law, R. et al. Cooperativity in forced unfolding of tandem spectrin repeats. Biophys. J. 84, 533\u2013544 (2003).","journal-title":"Biophys. J."},{"key":"41569_CR18","doi-asserted-by":"publisher","first-page":"468","DOI":"10.1073\/pnas.98.2.468","volume":"98","author":"AF Oberhauser","year":"2001","unstructured":"Oberhauser, A. F., Hansma, P. K., Carrion-Vazquez, M. & Fernandez, J. M. Stepwise unfolding of titin under force-clamp atomic force microscopy. Proc. Natl. Acad. Sci. 98, 468\u2013472 (2001).","journal-title":"Proc. Natl. Acad. Sci."},{"key":"41569_CR19","unstructured":"Press, W. H., Teukolsky, S. A., Vetterling, W. T. & Flannery, B. P. In The Art of Scientific Computing 647\u2013649 (Cambridge University Press, 2002)."},{"key":"41569_CR20","doi-asserted-by":"publisher","first-page":"2955","DOI":"10.1093\/hmg\/ddr199","volume":"20","author":"DM Henderson","year":"2011","unstructured":"Henderson, D. M., Belanto, J. J., Li, B., Heun-Johnson, H. & Ervasti, J. M. Internal deletion compromises the stability of dystrophin. Hum. Mol. Genet. 20, 2955\u201363 (2011).","journal-title":"Hum. Mol. Genet."},{"key":"41569_CR21","doi-asserted-by":"publisher","first-page":"1545","DOI":"10.2353\/ajpath.2009.090052","volume":"175","author":"JV Welser","year":"2009","unstructured":"Welser, J. V. et al. Myotendinous junction defects and reduced force transmission in mice that lack alpha7 integrin and utrophin. Am. J. Pathol. 175, 1545\u20131554 (2009).","journal-title":"Am. J. Pathol."},{"key":"41569_CR22","doi-asserted-by":"publisher","first-page":"883","DOI":"10.1083\/jcb.136.4.883","volume":"136","author":"AE Deconinck","year":"1997","unstructured":"Deconinck, A. E. et al. Postsynaptic abnormalities at the neuromuscular junctions of utrophin-deficient mice. J. Cell Biol. 136, 883\u201394 (1997).","journal-title":"J. Cell Biol."},{"key":"41569_CR23","doi-asserted-by":"publisher","first-page":"871","DOI":"10.1083\/jcb.136.4.871","volume":"136","author":"RM Grady","year":"1997","unstructured":"Grady, R. M., Merlie, J. P. & Sanes, J. R. Subtle neuromuscular defects in utrophin-deficient mice. J. Cell Biol. 136, 871\u201382 (1997).","journal-title":"J. Cell Biol."},{"key":"41569_CR24","doi-asserted-by":"publisher","first-page":"C1300","DOI":"10.1152\/ajpcell.2001.281.4.C1300","volume":"281","author":"AO Gramolini","year":"2001","unstructured":"Gramolini, A. O., B\u00e9langer, G., Thompson, J. M., Chakkalakal, J. V. & Jasmin, B. J. Increased expression of utrophin in a slow vs. a fast muscle involves posttranscriptional events. Am. J. Physiol. Cell Physiol. 281, C1300\u2013C1309 (2001).","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"41569_CR25","doi-asserted-by":"publisher","first-page":"13914","DOI":"10.1074\/jbc.M809743200","volume":"284","author":"Y Zhu","year":"2009","unstructured":"Zhu, Y., Bogomolovas, J., Labeit, S. & Granzier, H. Single molecule force spectroscopy of the cardiac titin N2B element: effects of the molecular chaperone alphaB-crystallin with disease-causing mutations. J. Biol. Chem. 284, 13914\u201313923 (2009).","journal-title":"J. Biol. Chem."},{"key":"41569_CR26","doi-asserted-by":"publisher","first-page":"105","DOI":"10.1016\/S0005-2728(03)00029-X","volume":"1604","author":"MSZ Kellermayer","year":"2003","unstructured":"Kellermayer, M. S. Z., Bustamante, C. & Granzier, H. L. Mechanics and structure of titin oligomers explored with atomic force microscopy. Biochim. Biophys. Acta 1604, 105\u2013114 (2003).","journal-title":"Biochim. Biophys. Acta"},{"key":"41569_CR27","doi-asserted-by":"publisher","first-page":"124","DOI":"10.1016\/S0014-5793(00)01704-X","volume":"476","author":"PF Lenne","year":"2000","unstructured":"Lenne, P. F., Raae, A. J., Altmann, S. M., Saraste, M. & Horber, J. K. States and transitions during forced unfolding of a single spectrin repeat. FEBS Lett. 476, 124\u2013128 (2000).","journal-title":"FEBS Lett."},{"key":"41569_CR28","doi-asserted-by":"publisher","first-page":"1085","DOI":"10.1016\/S0969-2126(02)00808-0","volume":"10","author":"SM Altmann","year":"2002","unstructured":"Altmann, S. M. et al. Pathways and intermediates in forced unfolding of spectrin repeats. Structure 10, 1085\u201396 (2002).","journal-title":"Structure"},{"key":"41569_CR29","doi-asserted-by":"publisher","first-page":"817","DOI":"10.1016\/j.jmb.2004.11.021","volume":"345","author":"L Li","year":"2005","unstructured":"Li, L., Huang, H. H.-L., Badilla, C. L. & Fernandez, J. M. Mechanical Unfolding Intermediates Observed by Single-molecule Force Spectroscopy in a Fibronectin Type III Module. J. Mol. Biol. 345, 817\u2013826 (2005).","journal-title":"J. Mol. Biol."},{"key":"41569_CR30","doi-asserted-by":"publisher","first-page":"392","DOI":"10.1016\/j.jsb.2014.03.011","volume":"186","author":"A Nicolas","year":"2014","unstructured":"Nicolas, A., Delalande, O., Hubert, J.-F. & Le Rumeur, E. The spectrin family of proteins: a unique coiled-coil fold for various molecular surface properties. J. Struct. Biol. 186, 392\u2013401 (2014).","journal-title":"J. Struct. Biol."},{"key":"41569_CR31","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1016\/0014-5793(95)00398-S","volume":"369","author":"SJ Winder","year":"1995","unstructured":"Winder, S. J., Gibson, T. J. & Kendrick-Jones, J. Dystrophin and utrophin: the missing links! FEBS Lett. 369, 27\u201333 (1995).","journal-title":"FEBS Lett."},{"key":"41569_CR32","doi-asserted-by":"publisher","first-page":"10745","DOI":"10.1021\/acsnano.6b03314","volume":"10","author":"AC Chang","year":"2016","unstructured":"Chang, A. C. et al. Single Molecule Force Measurements in Living Cells Reveal a Minimally Tensioned Integrin State. ACS Nano 10, 10745\u201310752 (2016).","journal-title":"ACS Nano"},{"key":"41569_CR33","doi-asserted-by":"publisher","first-page":"1254211","DOI":"10.1126\/science.1254211","volume":"346","author":"CD Buckley","year":"2014","unstructured":"Buckley, C. D. et al. Cell adhesion. The minimal cadherin-catenin complex binds to actin filaments under force. Science 346, 1254211 (2014).","journal-title":"Science"},{"key":"41569_CR34","first-page":"53","volume":"2","author":"B Charvet","year":"2012","unstructured":"Charvet, B., Ruggiero, F. & Le Guellec, D. The development of the myotendinous junction. A review. Muscles. Ligaments Tendons J. 2, 53\u201363 (2012).","journal-title":"Muscles. Ligaments Tendons J."},{"key":"41569_CR35","doi-asserted-by":"publisher","first-page":"194","DOI":"10.1006\/jsbi.2002.4468","volume":"137","author":"WA Linke","year":"2002","unstructured":"Linke, W. A. et al. PEVK domain of titin: an entropic spring with actin-binding properties. J. Struct. Biol. 137, 194\u2013205 (2002).","journal-title":"J. Struct. Biol."},{"key":"41569_CR36","doi-asserted-by":"publisher","first-page":"7101","DOI":"10.1073\/pnas.88.16.7101","volume":"88","author":"K Wang","year":"1991","unstructured":"Wang, K., McCarter, R., Wright, J., Beverly, J. & Ramirez-Mitchell, R. Regulation of skeletal muscle stiffness and elasticity by titin isoforms: a test of the segmental extension model of resting tension. Proc. Natl. Acad. Sci. USA 88, 7101\u20135 (1991).","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"41569_CR37","doi-asserted-by":"publisher","first-page":"930","DOI":"10.1103\/PhysRevLett.56.930","volume":"56","author":"Q Binnig","year":"1986","unstructured":"Binnig, Q. & Gerber. Atomic force microscope. Phys. Rev. Lett. 56, 930\u2013933 (1986).","journal-title":"Phys. Rev. Lett."},{"key":"41569_CR38","doi-asserted-by":"publisher","first-page":"3967","DOI":"10.1063\/1.1150021","volume":"70","author":"JE Sader","year":"1999","unstructured":"Sader, J. E., Chon, J. W. M. & Mulvaney, P. Calibration of rectangular atomic force microscope cantilevers. Rev. Sci. Instrum. 70, 3967 (1999).","journal-title":"Rev. Sci. Instrum."},{"key":"41569_CR39","doi-asserted-by":"publisher","first-page":"1236","DOI":"10.1139\/y86-209","volume":"64","author":"RB Stein","year":"1986","unstructured":"Stein, R. B. & Gordon, T. Nonlinear stiffness\u2013force relationships in whole mammalian skeletal muscles. Can. J. Physiol. Pharmacol. 64, 1236\u201344 (1986).","journal-title":"Can. J. Physiol. Pharmacol."},{"key":"41569_CR40","doi-asserted-by":"publisher","first-page":"819","DOI":"10.1002\/mus.880110804","volume":"11","author":"T Gordon","year":"1988","unstructured":"Gordon, T. & Stein, R. B. Comparison of force and stiffness in normal and dystrophic mouse muscles. Muscle Nerve 11, 819\u2013827 (1988).","journal-title":"Muscle Nerve"},{"key":"41569_CR41","doi-asserted-by":"publisher","unstructured":"Cohen, J. Statistical Power Analysis for the Behavioral Sciences, https:\/\/doi.org\/10.4324\/9780203771587 (Routledge, 1988).","DOI":"10.4324\/9780203771587"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-41569-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-41569-4","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-41569-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,17]],"date-time":"2022-12-17T08:09:54Z","timestamp":1671264594000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-41569-4"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,3,26]]},"references-count":41,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2019,12]]}},"alternative-id":["41569"],"URL":"https:\/\/doi.org\/10.1038\/s41598-019-41569-4","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,3,26]]},"assertion":[{"value":"21 September 2017","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"12 March 2019","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"26 March 2019","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"5210"}}