Calmodulin Binding and Inhibition of Cardiac Muscle Calcium Release Channel (Ryanodine Receptor)

Balshaw, David M.; Xu, Le; Yamaguchi, Naohiro; Pasek, Daniel A.; Meissner, Gerhard

doi:10.1074/jbc.m010771200

Cited by 202 publications

(275 citation statements)

References 33 publications

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“…Amino acids 3615 -3628 contact the carboxy lobe of CaM, whereas amino acids 3628 -3637 bind the amino lobe of CaM. In cardiac muscle, CaM shifts the Ca 2þ -dependence of RyR2 activation to higher Ca 2þ concentrations and hence decreases the RyR2 opening at all Ca 2þ concentrations (Balshaw et al 2001;Yamaguchi et al 2003). Recently reduced affinity for CaM binding to RyR2 with PKA phosphorylation was found in a CPVT-associated mouse model (Arg2474Ser), resulting in spontaneous local Ca 2þ release events leading to lethal arrythmias (Xu et al 2010).…”

Section: Calmodulinmentioning

confidence: 99%

Ryanodine Receptors: Structure, Expression, Molecular Details, and Function in Calcium Release

Lanner¹,

Georgiou²,

Joshi³

et al. 2010

Cold Spring Harbor Perspectives in Biology

658

572

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Ryanodine receptors (RyRs) are located in the sarcoplasmic/endoplasmic reticulum membrane and are responsible for the release of Ca 2þ from intracellular stores during excitation-contraction coupling in both cardiac and skeletal muscle. RyRs are the largest known ion channels (.2MDa) and exist as three mammalian isoforms (RyR 1 -3), all of which are homotetrameric proteins that interact with and are regulated by phosphorylation, redox modifications, and a variety of small proteins and ions. Most RyR channel modulators interact with the large cytoplasmic domain whereas the carboxy-terminal portion of the protein forms the ion-conducting pore. Mutations in RyR2 are associated with human disorders such as catecholaminergic polymorphic ventricular tachycardia whereas mutations in RyR1 underlie diseases such as central core disease and malignant hyperthermia. This chapter examines the current concepts of the structure, function and regulation of RyRs and assesses the current state of understanding of their roles in associated disorders.

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Section: Calmodulinmentioning

confidence: 99%

Ryanodine Receptors: Structure, Expression, Molecular Details, and Function in Calcium Release

Lanner¹,

Georgiou²,

Joshi³

et al. 2010

Cold Spring Harbor Perspectives in Biology

658

572

View full text Add to dashboard Cite

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“…RyR2 cDNA was kindly provided by Dr. Junichi Nakai of RIKEN Brain Science Institute (Wako, Japan). Unlabeled CaM and 35 S-CaM were prepared as described previously (10).…”

Section: Materials-[mentioning

confidence: 99%

“…Direct CaM binding inhibits all three RyR isoforms at [Ca 2ϩ ] Ͼ 1 M, whereas at [Ca 2ϩ ] Ͻ 1 M RyR1 and RyR3 are activated, but RyR2 is inhibited by CaM (6,7). 35 S-CaM binding to sarcoplasmic reticulum vesicles (8,9) and purified RyR1 and RyR2 (10) showed that the two receptor isoforms bind one molecule of CaM per RyR subunit in the absence and presence of Ca 2ϩ . CaM protection of trypsin digestion of RyR1 (11) and site-directed mutagenesis of RyR1 (12) and RyR2 (13) demonstrated that the two RyRs have a single CaM regulatory binding domain (CaMBD) (aa 3614 -3643 in RyR1) for the Ca 2ϩ -free and Ca 2ϩ -bound (Ca 2ϩ -CaM) forms of CaM.…”

mentioning

confidence: 99%

Different Regions in Skeletal and Cardiac Muscle Ryanodine Receptors Are Involved in Transducing the Functional Effects of Calmodulin

Yamaguchi

Evans

et al. 2004

Journal of Biological Chemistry

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Calmodulin (CaM) inhibits the skeletal muscle ryanodine receptor-1 (RyR1) and cardiac muscle RyR2 at micromolar Ca 2؉ but activates RyR1 and inhibits RyR2 at submicromolar Ca 2؉ by binding to a single, highly conserved CaM-binding site. To identify regions responsible for the differential regulation of RyR1 and RyR2 by CaM, we generated chimeras encompassing and flanking the CaM-binding domain. We found that the ex- (8, 9) and purified RyR1 and RyR2 (10) showed that the two receptor isoforms bind one molecule of CaM per RyR subunit in the absence and presence of Ca 2ϩ . CaM protection of trypsin digestion of RyR1 (11) and site-directed mutagenesis of RyR1 (12) and RyR2 (13) demonstrated that the two RyRs have a single CaM regulatory binding domain (CaMBD) (aa 3614 -3643 in RyR1) for the Ca 2ϩ -free and Ca 2ϩ -bound (Ca 2ϩ -CaM) forms of CaM. The CaM-binding domain is highly conserved among the mammalian isoforms, yet corresponding mutations in the RyR1 and RyR2 CaM-binding domain resulted in a different response to CaM in 35 S-CaM binding and functional measurements (13). This result suggests that other isoform-specific regions are responsible for the differential modulation of RyR1 and RyR2 by CaM.To test the hypothesis of an involvement of isoform-specific regions in regulating RyR1 and RyR2 by CaM, we constructed a series of RyR1/RyR2 chimeras and determined their CaMbinding properties and modulation by CaM. The results suggest that multiple regions are involved in transducing the functional effects of CaM in RyR1 and RyR2. The present study focused on the role of one of these regions. We found that substitution of five amino acid residues in RyR2 with those of RyR1 eliminated CaM inhibition but not CaM binding at [Ca 2ϩ ] Ͻ 1 M.

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“…Recently, mutations in CaM have also been shown to cause CPVT 31, 32, 33. Considering CaM binding to the cytosolic surface of RyR2 has been reported to contribute to inactivation of RyR2,17, 18, 19, 20 we tested whether arrhythmogenic mutations of CaM also act by shortening RyR2 refractoriness. Ca imaging was performed in permeabilized cardiac myocytes (cytosolic Ca buffered at 120 nmol/L) supplemented with different CaM protein variants 34, 35.…”

Section: Resultsmentioning

confidence: 99%

“…In cardiac cells, most CaM is bound to RyR2 and inhibits its activity in a Ca‐dependent manner 17, 18, 19, 20. Such Ca dependent binding would be expected to inhibit RyR2 specifically after Ca release lasting late into the diastolic period.…”

mentioning

confidence: 99%

Gene Transfer of Engineered Calmodulin Alleviates Ventricular Arrhythmias in a Calsequestrin‐Associated Mouse Model of Catecholaminergic Polymorphic Ventricular Tachycardia

Liu

Walton

et al. 2018

JAHA

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BackgroundCatecholaminergic polymorphic ventricular tachycardia (CPVT) is a familial arrhythmogenic syndrome characterized by sudden death. There are several genetic forms of CPVT associated with mutations in genes encoding the cardiac ryanodine receptor (RyR2) and its auxiliary proteins including calsequestrin (CASQ2) and calmodulin (CaM). It has been suggested that impairment of the ability of RyR2 to stay closed (ie, refractory) during diastole may be a common mechanism for these diseases. Here, we explore the possibility of engineering CaM variants that normalize abbreviated RyR2 refractoriness for subsequent viral‐mediated delivery to alleviate arrhythmias in non–CaM‐related CPVT.Methods and ResultsTo that end, we have designed a CaM protein (GSH‐M37Q; dubbed as therapeutic CaM or T‐CaM) that exhibited a slowed N‐terminal Ca dissociation rate and prolonged RyR2 refractoriness in permeabilized myocytes derived from CPVT mice carrying the CASQ2 mutation R33Q. This T‐CaM was introduced to the heart of R33Q mice through recombinant adeno‐associated viral vector serotype 9. Eight weeks postinfection, we performed confocal microscopy to assess Ca handling and recorded surface ECGs to assess susceptibility to arrhythmias in vivo. During catecholamine stimulation with isoproterenol, T‐CaM reduced isoproterenol‐promoted diastolic Ca waves in isolated CPVT cardiomyocytes. Importantly, T‐CaM exposure abolished ventricular tachycardia in CPVT mice challenged with catecholamines.ConclusionsOur results suggest that gene transfer of T‐CaM by adeno‐associated viral vector serotype 9 improves myocyte Ca handling and alleviates arrhythmias in a calsequestrin‐associated CPVT model, thus supporting the potential of a CaM‐based antiarrhythmic approach as a therapeutic avenue for genetically distinct forms of CPVT.

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Calmodulin Binding and Inhibition of Cardiac Muscle Calcium Release Channel (Ryanodine Receptor)

Cited by 202 publications

References 33 publications

Ryanodine Receptors: Structure, Expression, Molecular Details, and Function in Calcium Release

Ryanodine Receptors: Structure, Expression, Molecular Details, and Function in Calcium Release

Different Regions in Skeletal and Cardiac Muscle Ryanodine Receptors Are Involved in Transducing the Functional Effects of Calmodulin

Gene Transfer of Engineered Calmodulin Alleviates Ventricular Arrhythmias in a Calsequestrin‐Associated Mouse Model of Catecholaminergic Polymorphic Ventricular Tachycardia

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