+Open data
-Basic information
Entry | Database: PDB / ID: 7m3j | ||||||
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Title | Asymmetric Activation of the Calcium Sensing Receptor Homodimer | ||||||
Components | Extracellular calcium-sensing receptor | ||||||
Keywords | MEMBRANE PROTEIN / GPCR / calcium sensing receptor / negative allosteric modulator / family C GPCR | ||||||
Function / homology | Function and homology information bile acid secretion / chemosensory behavior / cellular response to peptide / response to fibroblast growth factor / cellular response to vitamin D / phosphatidylinositol phospholipase C activity / Class C/3 (Metabotropic glutamate/pheromone receptors) / calcium ion import / positive regulation of positive chemotaxis / fat pad development ...bile acid secretion / chemosensory behavior / cellular response to peptide / response to fibroblast growth factor / cellular response to vitamin D / phosphatidylinositol phospholipase C activity / Class C/3 (Metabotropic glutamate/pheromone receptors) / calcium ion import / positive regulation of positive chemotaxis / fat pad development / amino acid binding / cellular response to hepatocyte growth factor stimulus / branching morphogenesis of an epithelial tube / positive regulation of calcium ion import / regulation of calcium ion transport / cellular response to low-density lipoprotein particle stimulus / detection of calcium ion / anatomical structure morphogenesis / axon terminus / JNK cascade / positive regulation of vasoconstriction / chloride transmembrane transport / adenylate cyclase-inhibiting G protein-coupled receptor signaling pathway / ossification / response to ischemia / G protein-coupled receptor activity / cellular response to glucose stimulus / positive regulation of insulin secretion / intracellular calcium ion homeostasis / vasodilation / integrin binding / phospholipase C-activating G protein-coupled receptor signaling pathway / G alpha (i) signalling events / cellular response to hypoxia / G alpha (q) signalling events / basolateral plasma membrane / transmembrane transporter binding / positive regulation of ERK1 and ERK2 cascade / G protein-coupled receptor signaling pathway / apical plasma membrane / neuronal cell body / calcium ion binding / positive regulation of cell population proliferation / positive regulation of gene expression / protein kinase binding / cell surface / protein homodimerization activity / identical protein binding / plasma membrane Similarity search - Function | ||||||
Biological species | Homo sapiens (human) | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 4.1 Å | ||||||
Authors | Gao, Y. / Robertson, M.J. / Zhang, C. / Meyerowitz, J.G. / Panova, O. / Skiniotis, G. | ||||||
Funding support | United States, 1items
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Citation | Journal: Nature / Year: 2021 Title: Asymmetric activation of the calcium-sensing receptor homodimer. Authors: Yang Gao / Michael J Robertson / Sabrina N Rahman / Alpay B Seven / Chensong Zhang / Justin G Meyerowitz / Ouliana Panova / Fadil M Hannan / Rajesh V Thakker / Hans Bräuner-Osborne / Jesper ...Authors: Yang Gao / Michael J Robertson / Sabrina N Rahman / Alpay B Seven / Chensong Zhang / Justin G Meyerowitz / Ouliana Panova / Fadil M Hannan / Rajesh V Thakker / Hans Bräuner-Osborne / Jesper M Mathiesen / Georgios Skiniotis / Abstract: The calcium-sensing receptor (CaSR), a cell-surface sensor for Ca, is the master regulator of calcium homeostasis in humans and is the target of calcimimetic drugs for the treatment of parathyroid ...The calcium-sensing receptor (CaSR), a cell-surface sensor for Ca, is the master regulator of calcium homeostasis in humans and is the target of calcimimetic drugs for the treatment of parathyroid disorders. CaSR is a family C G-protein-coupled receptor that functions as an obligate homodimer, with each protomer composed of a Ca-binding extracellular domain and a seven-transmembrane-helix domain (7TM) that activates heterotrimeric G proteins. Here we present cryo-electron microscopy structures of near-full-length human CaSR in inactive or active states bound to Ca and various calcilytic or calcimimetic drug molecules. We show that, upon activation, the CaSR homodimer adopts an asymmetric 7TM configuration that primes one protomer for G-protein coupling. This asymmetry is stabilized by 7TM-targeting calcimimetic drugs adopting distinctly different poses in the two protomers, whereas the binding of a calcilytic drug locks CaSR 7TMs in an inactive symmetric configuration. These results provide a detailed structural framework for CaSR activation and the rational design of therapeutics targeting this receptor. | ||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | Molecule: MolmilJmol/JSmol |
-Downloads & links
-Download
PDBx/mmCIF format | 7m3j.cif.gz | 268.7 KB | Display | PDBx/mmCIF format |
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PDB format | pdb7m3j.ent.gz | 211.1 KB | Display | PDB format |
PDBx/mmJSON format | 7m3j.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 7m3j_validation.pdf.gz | 852 KB | Display | wwPDB validaton report |
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Full document | 7m3j_full_validation.pdf.gz | 859.3 KB | Display | |
Data in XML | 7m3j_validation.xml.gz | 44.4 KB | Display | |
Data in CIF | 7m3j_validation.cif.gz | 68.3 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/m3/7m3j ftp://data.pdbj.org/pub/pdb/validation_reports/m3/7m3j | HTTPS FTP |
-Related structure data
Related structure data | 23655MC 7m3eC 7m3fC 7m3gC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
#1: Protein | Mass: 101745.445 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Details: 1-16 is signaling sequence, 17-24 is FLAG epitope tag, 25-27 is an 3-alanine linker, the receptor sequence starts at residue Y28 that should be re-numbered to 20, and ends at V902 that ...Details: 1-16 is signaling sequence, 17-24 is FLAG epitope tag, 25-27 is an 3-alanine linker, the receptor sequence starts at residue Y28 that should be re-numbered to 20, and ends at V902 that should be re-numbered to 894. Source: (gene. exp.) Homo sapiens (human) / Gene: CASR, GPRC2A, PCAR1 / Cell (production host): sf9 / Production host: Spodoptera frugiperda (fall armyworm) / References: UniProt: P41180 #2: Polysaccharide | 2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose | Source method: isolated from a genetically manipulated source #3: Chemical | #4: Sugar | ChemComp-NAG / #5: Chemical | Has ligand of interest | Y | |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: inactive-state human extracellular calcium-sensing receptor complexed with negative allosteric modulator NPS-2143 Type: COMPLEX / Entity ID: #1 / Source: RECOMBINANT |
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Molecular weight | Value: 200 kDa/nm / Experimental value: NO |
Source (natural) | Organism: Homo sapiens (human) |
Source (recombinant) | Organism: Spodoptera frugiperda (fall armyworm) / Cell: sf9 |
Buffer solution | pH: 7.5 |
Specimen | Conc.: 7 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277 K |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / C2 aperture diameter: 70 µm |
Image recording | Electron dose: 60.9 e/Å2 / Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) |
-Processing
Software | Name: PHENIX / Version: 1.18.2_3874: / Classification: refinement | ||||||||||||||||||||||||
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EM software | Name: SerialEM / Category: image acquisition | ||||||||||||||||||||||||
CTF correction | Type: NONE | ||||||||||||||||||||||||
3D reconstruction | Resolution: 4.1 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 329093 / Symmetry type: POINT | ||||||||||||||||||||||||
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