How to open a Ryanodine Receptor

Petegem, Filip Van

doi:10.1038/cr.2016.106

Cited by 10 publications

(8 citation statements)

References 11 publications

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“…Immediate future work involves screening larger (50,000-compound) libraries, followed by lead development. Structure-function correlations enabled by biochemical, cell physiology, and spectroscopic studies in combination with recent advances in high-resolution RyR cryo-EM 25,58,[61][62][63][64][65][66] are expected to resolve the molecular mechanism of action for RyR modulators emerging from reports such as this one.…”

Section: Fkbp and Cam Binding As A Readout Of Hit Interaction With Thmentioning

confidence: 99%

RyR1-targeted drug discovery pipeline integrating FRET-based high-throughput screening and human myofiber dynamic Ca2+ assays

Rebbeck

Singh

Janicek

et al. 2020

Sci Rep

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Elevated cytoplasmic [Ca 2+ ] is characteristic in severe skeletal and cardiac myopathies, diabetes, and neurodegeneration, and partly results from increased Ca 2+ leak from sarcoplasmic reticulum stores via dysregulated ryanodine receptor (RyR) channels. Consequently, RyR is recognized as a high-value target for drug discovery to treat such pathologies. Using a FRET-based high-throughput screening assay that we previously reported, we identified small-molecule compounds that modulate the skeletal muscle channel isoform (RyR1) interaction with calmodulin and FK506 binding protein 12.6. Two such compounds, chloroxine and myricetin, increase FRET and inhibit [ 3 H]ryanodine binding to RyR1 at nanomolar Ca 2+. Both compounds also decrease RyR1 Ca 2+ leak in human skinned skeletal muscle fibers. Furthermore, we identified compound concentrations that reduced leak by > 50% but only slightly affected Ca 2+ release in excitation-contraction coupling, which is essential for normal muscle contraction. This report demonstrates a pipeline that effectively filters small-molecule RyR1 modulators towards clinical relevance. In striated muscle, contraction requires an intracellular Ca 2+-release event mediated by ryanodine receptors (RyR) that are embedded in the sarcoplasmic reticulum (SR) membrane. Dysregulation of skeletal (RyR1) and cardiac (RyR2) isoforms, via mutations or excess posttranslational modification, has been linked to severe muscle pathologies, including malignant hyperthermia (MH), central core disease, muscular dystrophy (MD), sarcopenia, catecholaminergic polymorphic ventricular tachycardia, heart failure, and more recently RyR2 has been recognized as a potentially significant contributor to diabetes and Alzheimer's disease 1-9. In most of these clinical indications, pathogenesis can be fueled by excess SR Ca 2+ "leak" via RyR under resting cellular conditions, which leads to toxic intracellular basal [Ca 2+ ] and insufficient SR Ca 2+ load. As a result, RyR is intensely studied as a therapeutic target. Indeed, the therapeutic potential of pharmaceutically targeting RyR1-mediated SR Ca 2+ leak in skeletal muscle has been shown in animal models of Duchenne MD, limb-girdle MD, and sarcopenia 5,10,11. The therapeutic potential of targeting RyR2-mediated SR Ca 2+ leak for treating heart failure and arrhythmia is also very well documented 12-16. Additionally, targeting RyR2 (which is abundant in the brain 17,18) may have therapeutic potential for treating neurodegenerative diseases. To introduce a systematic and efficient approach for identifying novel small-molecule chemical scaffolds with potential to mitigate RyR1 dysfunction, we developed and implemented a high-throughput screening (HTS) assay that uses fluorescence lifetime (FLT) detection of FRET 19. This assay was designed to identify compounds that bind to the RyR1 channel complex to allosterically correct its pathologically leaky state (without affecting normal channel function) 19. This FRET-based method is based on monitoring RyR binding of fluorescent...

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Section: Fkbp and Cam Binding As A Readout Of Hit Interaction With Thmentioning

confidence: 99%

RyR1-targeted drug discovery pipeline integrating FRET-based high-throughput screening and human myofiber dynamic Ca2+ assays

Rebbeck

Singh

Janicek

et al. 2020

Sci Rep

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“…The vast increase of knowledge presented by the cryo-EM studies in the past 2 years will be illuminated in the following sections of this review, and its implications will be discussed in the context of human disease. In particular, the newest structures offer a first glimpse on the dynamic mechanism of channel activation and gating (see Van Petegem 2016; Clarke and Hendrickson 2016). …”

Section: 2 Early Attempts To Determine Ryr Structurementioning

confidence: 99%

“…When this method is applied to the data collected in the study of RyR1 bound with various combinations of ligands (des Georges et al 2016), we indeed see that the channel in each sample assumes an entire continuum of states – albeit with different distributions – exhibiting a combination of wing motions, motions of the activation domain, and channel openings and closings (Dashti et al 2017). This analysis promises to lead to more profound insights into the nature and dynamics of channel activation and gating than obtainable from single reconstructions in open and closed states (see Van Petegem 2016). It may also be a well-suited tool to study how the numerous regulatory factors shift the equilibrium of states and modulate gating.…”

Section: 6 Mapping Of Channel Dynamics By Cryo-emmentioning

confidence: 99%

Ryanodine Receptor Structure and Function in Health and Disease

Santulli

Lewis

Georges

et al. 2018

Subcellular Biochemistry

111

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Ryanodine receptors (RyRs) are ubiquitous intracellular calcium (Ca2+) release channels required for the function of many organs including heart and skeletal muscle, synaptic transmission in the brain, pancreatic beta cell function, and vascular tone. In disease, defective function of RyRs due either to stress (hyperadrenergic and/or oxidative overload) or genetic mutations can render the channels leaky to Ca2+ and promote defective disease-causing signals as observed in heat failure, muscular dystrophy, diabetes mellitus, and neurodegerative disease. RyRs are massive structures comprising the largest known ion channel-bearing macromolecular complex and exceeding 3 million Daltons in molecular weight. RyRs mediate the rapid release of Ca2+ from the endoplasmic/sarcoplasmic reticulum (ER/SR) to stimulate cellular functions through Ca2+-dependent processes. Recent advances in single-particle cryogenic electron microscopy (cryo-EM) have enabled the determination of atomic-level structures for RyR for the first time. These structures have illuminated the mechanisms by which these critical ion channels function and interact with regulatory ligands. In the present chapter we discuss the structure, functional elements, gating and activation mechanisms of RyRs in normal and disease states.

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“…Ryanodine receptor (RyR) is a widely distributed endoplasmic reticulum (ER)-and sarcoplasmic reticulum (SR)-resident Ca 2+ channel [4]. Three isoforms of RyRs have been described, RyR1, 2 and 3.…”

Section: Introductionmentioning

confidence: 99%

Decrease of RyR2 in the prion infected cell line and in the brains of the scrapie infected mice models and the patients of human prion diseases

Shi

et al. 2018

Prion

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The levels of ryanodine receptors (RyRs) are usually increased in the brains of human Alzheimer disease (AD) and AD animal models. To evaluate the underlying alteration of brain RyRs in prion disease, scrapie infected cell line SMB-S15 and its infected mice were tested. RyR2 specific Western blots revealed markedly decreased RyR2 levels both in the cells and in the brains of infected mice. Assays of the brain samples of other scrapie (agents 139A and ME7) infected mice collected at different time-points during incubation period showed time-dependent decreases of RyR2. Immunofluorescent assays (IFA) verified that the expression of RyR2 locates predominantly in cytoplasm of SMB cells and overlapped with the neurons in the brain slices of mice. Furthermore, significant down-regulation of RyR2 was also detected in the postmortem cortical brains of the patients of various types of human prion diseases, including sporadic Creutzfeldt-Jakob disease (sCJD), fatal familial insomnia (FFI) and G114V-genetic CJD. Our data here propose the evidences of remarkably decreased brain RyR2 at terminal stages of both human prion diseases and prion infected rodent models. It also highlights that the therapeutic strategy with antagonist of RyRs in AD may not be suitable for prion disease.

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How to open a Ryanodine Receptor

Cited by 10 publications

References 11 publications

RyR1-targeted drug discovery pipeline integrating FRET-based high-throughput screening and human myofiber dynamic Ca2+ assays

RyR1-targeted drug discovery pipeline integrating FRET-based high-throughput screening and human myofiber dynamic Ca2+ assays

Ryanodine Receptor Structure and Function in Health and Disease

Decrease of RyR2 in the prion infected cell line and in the brains of the scrapie infected mice models and the patients of human prion diseases

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