ArticleProceedings of the National Academy of Sciences of the United States of America2001
Three-dimensional reconstruction of the recombinant type 3 ryanodine receptor and localization of its amino terminus.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers, 1 of them a synthesis that pooled it.
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Who cites it
26 citing papers in PubMed, 1 synthesis or guideline pooled it, 69 citations in OpenAlex.
- Calcium and arrhythmogenesis.Physiological reviews · 2007Pooled it
- Cryo-EM investigation of ryanodine receptor type 3.Nature communications · 2024Article
- Article
- A guide to the 3D structure of the ryanodine receptor type 1 by cryoEM.Protein science : a publication of the Protein Society · 2017Review
- Single-particle cryo-EM of the ryanodine receptor channel in an aqueous environment.European journal of translational myology · 2015Article
- Structure of glutaraldehyde cross-linked ryanodine receptor.Journal of structural biology · 2013Article
- Calmodulin-binding locations on the skeletal and cardiac ryanodine receptors.The Journal of biological chemistry · 2012Article
- FRET-based localization of fluorescent protein insertions within the ryanodine receptor type 1.PloS one · 2012Article
- Article
- The amino-terminal disease hotspot of ryanodine receptors forms a cytoplasmic vestibule.Nature · 2010Article
- Ryanodine receptors: structure, expression, molecular details, and function in calcium release.Cold Spring Harbor perspectives in biology · 2010Review
- Three-dimensional structure of recombinant type 1 inositol 1,4,5-trisphosphate receptor.The Biochemical journal · 2010Article
- Förster resonance energy transfer measurements of ryanodine receptor type 1 structure using a novel site-specific labeling method.PloS one · 2009Article
- Ryanodine receptor-mediated arrhythmias and sudden cardiac death.Pharmacology & therapeutics · 2009Review
- CLIC2-RyR1 interaction and structural characterization by cryo-electron microscopy.Journal of molecular biology · 2009Article
- Structural and functional characterization of ryanodine receptor-natrin toxin interaction.Biophysical journal · 2008Article
- The architecture of the DNA replication origin recognition complex in Saccharomyces cerevisiae.Proceedings of the National Academy of Sciences of the United States of America · 2008Article
- Subnanometer-resolution electron cryomicroscopy-based domain models for the cytoplasmic region of skeletal muscle RyR channel.Proceedings of the National Academy of Sciences of the United States of America · 2008Article
- ARVC-related mutations in divergent region 3 alter functional properties of the cardiac ryanodine receptor.Biophysical journal · 2008Article
- Localization of PKA phosphorylation site, Ser(2030), in the three-dimensional structure of cardiac ryanodine receptor.The Biochemical journal · 2008Article
Corrections and comments
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Authors and funding
6 authors at 4 institutions in 2 countries.
Funding
Abstract
Recombinant type 3 ryanodine receptor (RyR3) has been purified in quantities sufficient for structural characterization by cryoelectron microscopy and three-dimensional (3D) reconstruction. Two cDNAs were prepared and expressed in HEK293 cells, one encoding the wild-type RyR3 and the other encoding RyR3 containing glutathione S-transferase (GST) fused to its amino terminus (GST-RyR3). RyR3 was purified from detergent-solubilized transfected cells by affinity chromatography using 12.6-kDa FK506-binding protein in the form of a GST fusion as the affinity ligand. Purification of GST-RyR3 was achieved by affinity chromatography by using glutathione-Sepharose. Purified recombinant RyR3 and GST-RyR3 proteins exhibited high-affinity [(3)H]ryanodine binding that was sensitive to activation by Ca(2+) and caffeine and to inhibition by Mg(2+). 3D reconstructions of both recombinant RyR3 and GST-RyR3 appeared very similar to that of the native RyR3 purified from bovine diaphragm. Comparison of the 3D reconstructions of RyR3 and GST-RyR3 revealed that the GST domains and, hence, the amino termini of the RyR3 subunits are located in the "clamp" structures that form the corners of the square-shaped cytoplasmic region of homotetrameric RyR3. This study describes the 3D reconstruction of a recombinant ryanodine receptor and it demonstrates the potential of this technology for characterizing functional and structural perturbations introduced by site-directed mutagenesis.
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