RyR2 disease mutations at the C-terminal domain intersubunit interface alter closed-state stability and channel activation.
Ryanodine receptors (RyRs) are ion channels that mediate the release of Ca2+ from the sarcoplasmic reticulum/endoplasmic reticulum, mutations of which are implicated in a number of human diseases. The adjacent C-terminal domains (CTDs) of cardiac RyR (RyR2) interact with each other to form a ring-like tetrameric structure with the intersubunit interface undergoing dynamic changes during channel gating. This mobile CTD intersubunit interface harbors many disease-associated mutations. However, the mechanisms of action of these mutations and the role of CTD in channel function are not well understood. Here, we assessed the impact of CTD disease-associated mutations P4902S, P4902L, E4950K, and G4955E on Ca2+- and caffeine-mediated activation of RyR2. The G4955E mutation dramatically increased both the Ca2+-independent basal activity and Ca2+-dependent activation of [3H]ryanodine binding to RyR2. The P4902S and E4950K mutations also increased Ca2+ activation but had no effect on the basal activity of RyR2. All four disease mutations increased caffeine-mediated activation of RyR2 and reduced the threshold for activation and termination of spontaneous Ca2+ release. G4955D dramatically increased the basal activity of RyR2, whereas G4955K mutation markedly suppressed channel activity. Similarly, substitution of P4902 with a negatively charged residue (P4902D), but not a positively charged residue (P4902K), also dramatically increased the basal activity of RyR2. These data suggest that electrostatic interactions are involved in stabilizing the CTD intersubunit interface and that the G4955E disease mutation disrupts this interface, and thus the stability of the closed state. Our studies shed new insights into the mechanisms of action of RyR2 CTD disease mutations.
['Guo|Wenting|W|', 'Wei|Jinhong|J|', 'Estillore|John Paul|JP|', 'Zhang|Lin|L|', 'Wang|Ruiwu|R|', 'Sun|Bo|B|', 'Chen|S R Wayne|SRW|']
D000818:Animals D002110:Caffeine D002118:Calcium D004252:DNA Mutational Analysis D057809:HEK293 Cells D006801:Humans D015640:Ion Channel Gating D051379:Mice D009154:Mutation D011485:Protein Binding D000072417:Protein Domains D021122:Protein Subunits D012433:Ryanodine D019837:Ryanodine Receptor Calcium Release Channel D014316:Tritium
C-terminal domain ryanodine receptor channel gating Ca(2+) activation Ca(2+) release [(3)H]ryanodine binding cardiac arrythmias single-channel recordings