{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,5,2]],"date-time":"2025-05-02T04:15:23Z","timestamp":1746159323389,"version":"3.40.4"},"reference-count":36,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2014,2,5]],"date-time":"2014-02-05T00:00:00Z","timestamp":1391558400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001700","name":"Ministry of Education, Culture, Sport, Science and Technology of Japan","doi-asserted-by":"crossref","award":["23659112"],"award-info":[{"award-number":["23659112"]}],"id":[{"id":"10.13039\/501100001700","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pacing Clinical Electrophis"],"published-print":{"date-parts":[[2014,7]]},"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>A KCNE1 polymorphism, D85N, causes long QT syndrome (LQTS) with a decrease in the slowly activating delayed\u2010rectifier K<jats:sup>+<\/jats:sup>channel current (I<jats:sub>Ks<\/jats:sub>). We examined impacts of D85N polymorphism on KCNE1 protein stability and functions, and tested the ability of various drugs to modify them.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>KCNE1\u2010D85N or the wild\u2010type protein was coexpressed in COS7 cells with KCNQ1 to form K<jats:sup>+<\/jats:sup>channels. Expression, degradation, and intracellular localization of KCNE1 proteins, as well as the currents conferred by KCNQ1\/KCNE1 complexes, were determined using immunoblots, immunofluorescence, and patch\u2010clamp techniques.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The protein level of KCNE1\u2010D85N was lower than that of the wild\u2010type, in spite of the comparable levels of their mRNA. KCNE1\u2010D85N was highly ubiquitinated and rapidly degraded as compared to the wild\u2010type; a proteasome inhibitor, MG132, inhibited its degradation and increased its steady\u2010state level. Both KCNE1\u2010D85N and the wild\u2010type proteins were co\u2010immunoprecipitated with KCNQ1. Immunofluorescent signals of KCNE1\u2010D85N accumulated in the endoplasmic reticulum and Golgi apparatus, with reduced levels on the cell membrane. Patch\u2010clamp experiments demonstrated that the membrane current corresponding to I<jats:sub>Ks<\/jats:sub>was much smaller in cells expressing KCNE1\u2010D85N than in those expressing the wild\u2010type. Verapamil (0.5\u201310 \u03bcM) increased the protein level of KCNE1\u2010D85N, decreased its ubiquitination, slowed its degradation, and enhanced KCNQ1\/KCNE1\u2010D85N channel currents. Pretreatment with amiodarone abolished these effects of verapamil.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>KCNE1\u2010D85N is less stable than the wild\u2010type protein, and is rapidly degraded through the ubiquitin\u2010proteasome system. Verapamil may be of a therapeutic value in LQTS patients via preventing degradation of KCNE1\u2010D85N.<\/jats:p><\/jats:sec>","DOI":"10.1111\/pace.12360","type":"journal-article","created":{"date-parts":[[2014,2,5]],"date-time":"2014-02-05T21:27:17Z","timestamp":1391635637000},"page":"853-863","source":"Crossref","is-referenced-by-count":5,"title":["Instability of KCNE1\u2010D85N that Causes Long QT Syndrome: Stabilization by Verapamil"],"prefix":"10.1111","volume":"37","author":[{"given":"SHINJI","family":"SAKATA","sequence":"first","affiliation":[{"name":"Department of Pediatrics Faculty of Medicine Tottori University Yonago Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"YASUTAKA","family":"KURATA","sequence":"additional","affiliation":[{"name":"Department of Physiology Kanazawa Medical University Uchinada\u2010machi Kahoku\u2010gun Ishikawa 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