Roles of the NH2-terminal Domains of Cardiac Ryanodine Receptor in Ca2+ Release Activation and Termination

Liu, Yingjie; Sun, Bo; Xiao, Zhichao; Wang, Ruiwu; Guo, Wenting; Zhang, Joe Z.; Mi, Tao; Wang, Yundi; Jones, Peter P.; Petegem, Filip Van; Chen, S.R. Wayne

doi:10.1074/jbc.m114.618827

Cited by 17 publications

(21 citation statements)

References 44 publications

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“…The HEK293 cell endogenous SERCA2b operates at constant, maximum capacity under the perfusion conditions chosen, and CaM does not regulate SERCA2b as also evident from the a control experiment with RyR2‐ΔCaMBD (Fig. C) . The differences in steady‐state Ca 2+ load with no CaM, with CaM mutant and with CaM‐WT were evaluated using two‐way ANOVA (CaM presence vs. perfusion condition) with a Tukey's multiple comparisons correction, P < 0.05 considered significant.…”

Section: Methodsmentioning

confidence: 98%

“…Measurements of permeabilized HEK293 single cell steady‐state ER Ca 2+ load during perfusion with sustained 1 μ m [Ca 2+ ] free was done as previously described with modifications . Briefly, RyR2‐expressing HEK293 cells were permeabilized by perfusion with 0.25 g·L −1 saponin in Ca 2+ free intracellular‐like medium (ICM, 125 m m KCl, 19 m m NaCl, 10 m m HEPES, 2 m m ATP, 2 m m MgCl 2 , and 50 μ m EGTA at pH 7.4) for 1–2 min.…”

Section: Methodsmentioning

confidence: 99%

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Diminished inhibition and facilitated activation of RyR2‐mediated Ca²⁺ release is a common defect of arrhythmogenic calmodulin mutations

et al. 2019

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A number of calmodulin (CaM) mutations cause severe cardiac arrhythmias, but their arrhythmogenic mechanisms are unclear. While some of the arrhythmogenic CaM mutations have been shown to impair CaM‐dependent inhibition of intracellular Ca2+ release through the ryanodine receptor type 2 (RyR2), the impact of a majority of these mutations on RyR2 function is unknown. Here, we investigated the effect of 14 arrhythmogenic CaM mutations on the CaM‐dependent RyR2 inhibition. We found that all the arrhythmogenic CaM mutations tested diminished CaM‐dependent inhibition of RyR2‐mediated Ca2+ release and increased store‐overload induced Ca2+ release (SOICR) in HEK293 cells. Moreover, all the arrhythmogenic CaM mutations tested either failed to inhibit or even promoted RyR2‐mediated Ca2+ release in permeabilized HEK293 cells with elevated cytosolic Ca2+, which was markedly different from the inhibitory action of CaM wild‐type. The CaM mutations also altered the Ca2+‐dependency of CaM binding to the RyR2 CaM‐binding domain. These results demonstrate that diminished inhibition, and even facilitated activation, of RyR2–mediated Ca2+ release is a common defect of arrhythmogenic CaM mutations.

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Section: Methodsmentioning

confidence: 98%

Section: Methodsmentioning

confidence: 99%

Diminished inhibition and facilitated activation of RyR2‐mediated Ca²⁺ release is a common defect of arrhythmogenic calmodulin mutations

et al. 2019

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“…After washing for 5 min three times, the bound antibodies were detected using an enhanced chemiluminescence kit from Pierce. The intensity of each band was determined from its intensity profile obtained by ImageQuant LAS 4000 (GE Healthcare Life Sciences) and analyzed by using the ImageJ software (36).…”

Section: Generation Of Stable Inducible Cell Lines Expressing Wt Andmentioning

confidence: 99%

Enhanced Cytosolic Ca2+ Activation Underlies a Common Defect of Central Domain Cardiac Ryanodine Receptor Mutations Linked to Arrhythmias

Xiao

Guo

Sun

et al. 2016

Journal of Biological Chemistry

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Edited by Roger ColbranRecent three-dimensional structural studies reveal that the central domain of ryanodine receptor (RyR) serves as a transducer that converts long-range conformational changes into the gating of the channel pore. Interestingly, the central domain encompasses one of the mutation hotspots (corresponding to amino acid residues 3778 -4201) that contains a number of cardiac RyR (RyR2) mutations associated with catecholaminergic polymorphic ventricular tachycardia (CPVT) and atrial fibrillation (AF). However, the functional consequences of these central domain RyR2 mutations are not well understood. To gain insights into the impact of the mutation and the role of the central domain in channel function, we generated and characterized eight disease-associated RyR2 mutations in the central domain. We found that all eight central domain RyR2 mutations enhanced the Ca 2؉

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“…After washing for 10 min three times, the bound antibodies were detected using an enhanced chemiluminescence kit from Pierce. The intensity of each band was determined from its intensity profile obtained by ImageQuant LAS 4000 (GE Healthcare/Life Sciences), analyzed by ImageJ software, and normalized to that of ␤-actin (40).…”

Section: Caffeine-induced Ca 2ϩmentioning

confidence: 99%

The Cytoplasmic Region of Inner Helix S6 Is an Important Determinant of Cardiac Ryanodine Receptor Channel Gating

Sun

Guo

Tian

et al. 2016

Journal of Biological Chemistry

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Edited by Roger ColbranThe ryanodine receptor (RyR) channel pore is formed by four S6 inner helices, with its intracellular gate located at the S6 helix bundle crossing region. The cytoplasmic region of the extended S6 helix is held by the U motif of the central domain and is thought to control the opening and closing of the S6 helix bundle. However, the functional significance of the S6 cytoplasmic region in channel gating is unknown. Here we assessed the role of the S6 cytoplasmic region in the function of cardiac RyR (RyR2) via structure-guided site-directed mutagenesis.

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Roles of the NH2-terminal Domains of Cardiac Ryanodine Receptor in Ca2+ Release Activation and Termination

Cited by 17 publications

References 44 publications

Diminished inhibition and facilitated activation of RyR2‐mediated Ca²⁺ release is a common defect of arrhythmogenic calmodulin mutations

Diminished inhibition and facilitated activation of RyR2‐mediated Ca²⁺ release is a common defect of arrhythmogenic calmodulin mutations

Enhanced Cytosolic Ca2+ Activation Underlies a Common Defect of Central Domain Cardiac Ryanodine Receptor Mutations Linked to Arrhythmias

The Cytoplasmic Region of Inner Helix S6 Is an Important Determinant of Cardiac Ryanodine Receptor Channel Gating

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Roles of the NH2-terminal Domains of Cardiac Ryanodine Receptor in Ca2+ Release Activation and Termination

Cited by 17 publications

References 44 publications

Diminished inhibition and facilitated activation of RyR2‐mediated Ca2+ release is a common defect of arrhythmogenic calmodulin mutations

Diminished inhibition and facilitated activation of RyR2‐mediated Ca2+ release is a common defect of arrhythmogenic calmodulin mutations

Enhanced Cytosolic Ca2+ Activation Underlies a Common Defect of Central Domain Cardiac Ryanodine Receptor Mutations Linked to Arrhythmias

The Cytoplasmic Region of Inner Helix S6 Is an Important Determinant of Cardiac Ryanodine Receptor Channel Gating

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Diminished inhibition and facilitated activation of RyR2‐mediated Ca²⁺ release is a common defect of arrhythmogenic calmodulin mutations

Diminished inhibition and facilitated activation of RyR2‐mediated Ca²⁺ release is a common defect of arrhythmogenic calmodulin mutations