Spontaneous Ca2+ release from intracellular stores is important for various physiological and pathological processes. In cardiac muscle cells, spontaneous store overload–induced Ca2+ release (SOICR) can result in Ca2+ waves, a major cause of ventricular tachyarrhythmias (VTs) and sudden death. The molecular mechanism underlying SOICR has been a mystery for decades. Here we show that a point mutation, E4872A, in the helix bundle crossing region (the proposed gate) of the cardiac ryanodine receptor (RyR2) completely abolishes luminal, but not cytosolic, Ca2+ activation of RyR2. The introduction of metal-binding histidines at this site converts RyR2 into a luminal Ni2+-gated channel. Mouse hearts harboring a heterozygous RyR2 mutation at this site (E4872Q) are resistant to SOICR and are completely protected against Ca2+-triggered VTs. These data show that the RyR2 gate directly senses luminal (store) Ca2+, explaining the regulation of RyR2 by luminal Ca2+, the initiation of Ca2+ waves and Ca2+-triggered arrhythmias. This newly identified store-sensing gate structure is conserved in all RyR and inositol 1,4,5-trisphosphate receptor isoforms.
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This work was supported by research grants from the Canadian Institutes of Health Research (CIHR) to H.J.D., D.P.T. and S.R.W.C., the US National Institutes of Health to J.C., L.-S.S., M.F. and S.R.W.C., the Heart and Stroke Foundation of Alberta to A.M.G., H.J.D. and S.R.W.C. and the Canada Foundation for Innovation (S.R.W.C.) and by the Heart and Stroke Foundation/Libin Cardiovascular Institute Professorship in Cardiovascular Research (S.R.W.C.). We are grateful for generous donations from the King family, the Howarth family and the Libin Cardiovascular Institute of Alberta. We are also grateful to W.C. Claycomb from the Louisiana State University Health Sciences Center for kindly providing the HL-1 cardiac cell line and to J. Lytton from the University of Calgary for the gift of the β-actin–specific antibody. W.C., X.Z. and X.T. are recipients of the Alberta Innovates-Health Solutions (AIHS) Studentship Award; P.P.J. is a recipient of AIHS and Heart and Stroke Foundation Fellowship Awards; M.L.O. is the recipient of a CIHR Fellowship Award; and H.J.D., D.P.T. and S.R.W.C. are recipients of AIHS Scientist Awards.
The authors declare no competing financial interests.
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Chen, W., Wang, R., Chen, B. et al. The ryanodine receptor store-sensing gate controls Ca2+ waves and Ca2+-triggered arrhythmias. Nat Med 20, 184–192 (2014). https://doi.org/10.1038/nm.3440
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