{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T17:28:19Z","timestamp":1771954099703,"version":"3.50.1"},"reference-count":66,"publisher":"Wiley","issue":"23","license":[{"start":{"date-parts":[[2021,11,9]],"date-time":"2021-11-09T00:00:00Z","timestamp":1636416000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["physoc.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Journal of Physiology"],"published-print":{"date-parts":[[2021,12]]},"abstract":"<jats:sec><jats:title>Abstract<\/jats:title><jats:p>Catecholaminergic polymorphic ventricular tachycardia type 1 (CPVT1) is a lethal genetic disease causing arrhythmias and sudden cardiac death in children and young adults and is linked to mutations in the cardiac ryanodine receptor (RyR2). The effects of CPVT1 mutations on RyR2 ion\u2010channel function are often investigated using purified recombinant RyR2 channels homozygous for the mutation. However, CPVT1 patients are heterozygous for the disease, so this approach does not reveal the true changes to RyR2 function across the entire RyR2 population of channels in the heart. We therefore investigated the native cardiac RyR2 single\u2010channel abnormalities in mice heterozygous for the CPVT1 mutation, V2475F(+\/\u2010)\u2010RyR2, and applied molecular modelling techniques to investigate the possible structural changes that could initiate any altered function. We observed that increased sensitivity of cardiac V2475F(+\/\u2010)\u2010RyR2 channels to both activating and inactivating levels of cytosolic Ca<jats:sup>2+<\/jats:sup>, plus attenuation of Mg<jats:sup>2+<\/jats:sup> inhibition, were the most marked changes. Severity of abnormality was not uniform across all channels, giving rise to multiple sub\u2010populations with differing functional characteristics. For example, 46% of V2475F(+\/\u2010)\u2010RyR2 channels exhibited reduced Mg<jats:sup>2+<\/jats:sup> inhibition and 23% were actually activated by Mg<jats:sup>2+<\/jats:sup>. Using homology modelling, we discovered that V2475 is situated at a hinge between two regions of the RyR2 helical domain 1 (HD1). Our model proposes that detrimental functional changes to RyR2 arise because mutation at this critical site reduces the angle between these regions. Our results demonstrate the necessity of characterising the total heterozygous population of CPVT1\u2010mutated channels in order to understand CPVT1 phenotypes in patients.\n<jats:boxed-text content-type=\"graphic\" position=\"anchor\"><jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" mimetype=\"image\/png\" position=\"anchor\" specific-use=\"enlarged-web-image\" xlink:href=\"graphic\/tjp14871-gra-0001-m.png\"><jats:alt-text>image<\/jats:alt-text><\/jats:graphic><\/jats:boxed-text><\/jats:p><\/jats:sec><jats:sec><jats:title>Key points<\/jats:title><jats:p><jats:list list-type=\"bullet\">\n<jats:list-item>\n<jats:p>RyR2 mutations can cause type\u20101 catecholaminergic polymorphic ventricular tachycardia (CPVT1), a lethal, autosomal\u2010dominant arrhythmic disease. However, the changes in RyR2 ion\u2010channel function that result from the many different patient mutations are rarely investigated in detail and often only recombinant RyR2, homozygous for the mutation, is studied. As CPVT1 is a heterozygous disease and the tetrameric RyR2 channels expressed in the heart will contain varying numbers of mutated monomers, we have investigated the range of RyR2 single\u2010channel abnormalities found in the hearts of mice heterozygous for the CPVT1 mutation, V2475F(+\/\u2010)\u2010RyR2.<\/jats:p>\n<\/jats:list-item>\n<jats:list-item>\n<jats:p>Specific alterations to ligand regulation of V2475F(+\/\u2010)\u2010RyR2 were observed. Multiple sub\u2010populations of channels exhibited varying degrees of abnormality. In particular, an increased sensitivity to activating and inactivating cytosolic [Ca<jats:sup>2+<\/jats:sup>], and reduced sensitivity to Mg<jats:sup>2+<\/jats:sup> inhibition were evident.<\/jats:p>\n<\/jats:list-item>\n<jats:list-item>\n<jats:p>Our results provide mechanistic insight into the changes to RyR2 gating that destabilise sarcoplasmic reticulum Ca<jats:sup>2+<\/jats:sup>\u2010release causing life\u2010threatening arrhythmias in V2475F(+\/\u2010)\u2010CPVT1 patients.<\/jats:p>\n<\/jats:list-item>\n<\/jats:list><\/jats:p><\/jats:sec>","DOI":"10.1113\/jp281707","type":"journal-article","created":{"date-parts":[[2021,10,22]],"date-time":"2021-10-22T07:35:00Z","timestamp":1634888100000},"page":"5179-5201","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["The V2475F CPVT1 mutation yields distinct RyR2 channel populations that differ in their responses to cytosolic Ca<sup>2+<\/sup> and 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