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Myocardial function can be modulated through sequestration of myosin motors into an auto-inhibited \u201csuper-relaxed\u201d state (SRX), which may be further stabilized by a structural state known as the \u201cinteracting heads motif\u201d (IHM). Here, we sought to determine whether hypo-contractility of DCM myocardium results from reduced function of individual myosin molecules or from decreased myosin availability to interact with actin due to increased IHM\/SRX stabilization. We used an established DCM myosin mutation, E525K, and characterized the biochemical and mechanical activity of wild-type and mutant human \u03b2-cardiac myosin constructs that differed in the length of their coiled-coil tail, which dictates their ability to form the IHM\/SRX state. We found that short-tailed myosin constructs exhibited low IHM\/SRX content, elevated actin-activated ATPase activity, and fast velocities in unloaded motility assays. Conversely, longer-tailed constructs exhibited higher IHM\/SRX content and reduced actomyosin ATPase and velocity. Our modeling suggests that reduced velocities may be attributed to IHM\/SRX-dependent sequestration of myosin heads. Interestingly, longer-tailed E525K mutants showed no apparent impact on velocity or actomyosin ATPase at low ionic strength but stabilized IHM\/SRX state at higher ionic strength. Therefore, the hypo-contractility observed in DCM may be attributable to reduced myosin head availability caused by enhanced IHM\/SRX stability in E525K mutants.<\/jats:p>","DOI":"10.1085\/jgp.202313522","type":"journal-article","created":{"date-parts":[[2024,4,18]],"date-time":"2024-04-18T02:20:53Z","timestamp":1713406853000},"update-policy":"https:\/\/doi.org\/10.1085\/jgp.crossmarkpolicy","source":"Crossref","is-referenced-by-count":16,"title":["Tail length and E525K dilated cardiomyopathy mutant alter human \u03b2-cardiac myosin super-relaxed state"],"prefix":"10.1085","volume":"156","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2618-1335","authenticated-orcid":true,"given":"Sebastian","family":"Duno-Miranda","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0155zta11","id-type":"ROR","asserted-by":"publisher"}],"name":"Cardiovascular Research Institute, University of Vermont 1 Department of Molecular Physiology and Biophysics, , Burlington, VT,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3032-6314","authenticated-orcid":true,"given":"Shane R.","family":"Nelson","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0155zta11","id-type":"ROR","asserted-by":"publisher"}],"name":"Cardiovascular Research Institute, University of Vermont 1 Department of Molecular Physiology and Biophysics, , Burlington, VT,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3891-0694","authenticated-orcid":true,"given":"David V.","family":"Rasicci","sequence":"additional","affiliation":[{"name":"Penn State College of Medicine 2 Department of Cellular and Molecular Physiology, , Hershey, PA,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5794-6927","authenticated-orcid":true,"given":"Skylar M.L.","family":"Bodt","sequence":"additional","affiliation":[{"name":"Penn State College of Medicine 2 Department of Cellular and Molecular Physiology, , Hershey, PA,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2135-5831","authenticated-orcid":true,"suffix":"Jr","given":"Joseph A.","family":"Cirilo","sequence":"additional","affiliation":[{"name":"Penn State College of Medicine 2 Department of Cellular and Molecular Physiology, , Hershey, PA,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8990-8968","authenticated-orcid":true,"given":"Duha","family":"Vang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"University of Minnesota 3 Department of Genetics, Cell Biology, and Development, , Minneapolis, MN,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9541-6994","authenticated-orcid":true,"given":"Sivaraj","family":"Sivaramakrishnan","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017zqws13","id-type":"ROR","asserted-by":"publisher"}],"name":"University of Minnesota 3 Department of Genetics, Cell Biology, and Development, , Minneapolis, MN,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3987-9019","authenticated-orcid":true,"given":"Christopher M.","family":"Yengo","sequence":"additional","affiliation":[{"name":"Penn State College of Medicine 2 Department of Cellular and Molecular Physiology, , Hershey, PA,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0071-8798","authenticated-orcid":true,"given":"David M.","family":"Warshaw","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0155zta11","id-type":"ROR","asserted-by":"publisher"}],"name":"Cardiovascular Research Institute, University of Vermont 1 Department of Molecular Physiology and Biophysics, , Burlington, VT,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2024,5,6]]},"reference":[{"key":"2025050520021383800_bib1","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.24634","article-title":"Effects of myosin variants on interacting-heads motif explain distinct hypertrophic and dilated cardiomyopathy phenotypes","volume":"6","author":"Alamo","year":"2017","journal-title":"Elife"},{"key":"2025050520021383800_bib2","doi-asserted-by":"publisher","first-page":"E8143","DOI":"10.1073\/pnas.1809540115","article-title":"Deciphering the super relaxed state of human \u03b2-cardiac myosin and the mode of action of mavacamten from myosin molecules to muscle fibers","volume":"115","author":"Anderson","year":"2018","journal-title":"Proc. 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