+Open data
-Basic information
Entry | Database: PDB / ID: 6wth | |||||||||||||||
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Title | Full-length human ENaC ECD | |||||||||||||||
Components |
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Keywords | MEMBRANE PROTEIN / sodium channel / blood pressure / epithelial / salt transport | |||||||||||||||
Function / homology | Function and homology information sensory perception of salty taste / Sensory perception of salty taste / neutrophil-mediated killing of bacterium / sensory perception of sour taste / leukocyte activation involved in inflammatory response / aldosterone metabolic process / cellular response to vasopressin / sperm principal piece / sodium channel complex / ligand-gated sodium channel activity ...sensory perception of salty taste / Sensory perception of salty taste / neutrophil-mediated killing of bacterium / sensory perception of sour taste / leukocyte activation involved in inflammatory response / aldosterone metabolic process / cellular response to vasopressin / sperm principal piece / sodium channel complex / ligand-gated sodium channel activity / epithelial fluid transport / sodium ion homeostasis / mucus secretion / artery smooth muscle contraction / renal system process / neutrophil activation involved in immune response / cellular response to aldosterone / multicellular organismal-level water homeostasis / cellular response to acidic pH / sodium ion import across plasma membrane / potassium ion homeostasis / intracellular sodium ion homeostasis / motile cilium / sodium channel activity / erythrocyte homeostasis / response to food / ciliary membrane / WW domain binding / sodium ion transport / sodium ion transmembrane transport / acrosomal vesicle / monoatomic ion channel activity / cytoplasmic vesicle membrane / multicellular organism growth / Stimuli-sensing channels / regulation of blood pressure / gene expression / response to xenobiotic stimulus / apical plasma membrane / external side of plasma membrane / extracellular exosome / nucleoplasm / plasma membrane / cytosol / cytoplasm Similarity search - Function | |||||||||||||||
Biological species | Homo sapiens (human) Mus musculus (house mouse) | |||||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.06 Å | |||||||||||||||
Authors | Posert, R. / Baconguis, I. / Noreng, S. / Bharadwaj, A. / Houser, A. | |||||||||||||||
Funding support | United States, 4items
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Citation | Journal: Elife / Year: 2020 Title: Molecular principles of assembly, activation, and inhibition in epithelial sodium channel. Authors: Sigrid Noreng / Richard Posert / Arpita Bharadwaj / Alexandra Houser / Isabelle Baconguis / Abstract: The molecular bases of heteromeric assembly and link between Na self-inhibition and protease-sensitivity in epithelial sodium channels (ENaCs) are not fully understood. Previously, we demonstrated ...The molecular bases of heteromeric assembly and link between Na self-inhibition and protease-sensitivity in epithelial sodium channels (ENaCs) are not fully understood. Previously, we demonstrated that ENaC subunits - α, β, and γ - assemble in a counterclockwise configuration when viewed from outside the cell with the protease-sensitive GRIP domains in the periphery (Noreng et al., 2018). Here we describe the structure of ENaC resolved by cryo-electron microscopy at 3 Å. We find that a combination of precise domain arrangement and complementary hydrogen bonding network defines the subunit arrangement. Furthermore, we determined that the α subunit has a primary functional module consisting of the finger and GRIP domains. The module is bifurcated by the α2 helix dividing two distinct regulatory sites: Na and the inhibitory peptide. Removal of the inhibitory peptide perturbs the Na site via the α2 helix highlighting the critical role of the α2 helix in regulating ENaC function. | |||||||||||||||
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 | 6wth.cif.gz | 278.2 KB | Display | PDBx/mmCIF format |
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PDB format | pdb6wth.ent.gz | 221.2 KB | Display | PDB format |
PDBx/mmJSON format | 6wth.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 6wth_validation.pdf.gz | 1.3 MB | Display | wwPDB validaton report |
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Full document | 6wth_full_validation.pdf.gz | 1.4 MB | Display | |
Data in XML | 6wth_validation.xml.gz | 57 KB | Display | |
Data in CIF | 6wth_validation.cif.gz | 85.6 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/wt/6wth ftp://data.pdbj.org/pub/pdb/validation_reports/wt/6wth | HTTPS FTP |
-Related structure data
Related structure data | 21896MC 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
-Amiloride-sensitive sodium channel subunit ... , 3 types, 3 molecules ABC
#1: Protein | Mass: 75780.531 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: SCNN1A, SCNN1 / Production host: Homo sapiens (human) / References: UniProt: P37088 |
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#2: Protein | Mass: 72728.891 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: SCNN1B / Production host: Homo sapiens (human) / References: UniProt: P51168 |
#3: Protein | Mass: 74352.984 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: SCNN1G / Production host: Homo sapiens (human) / References: UniProt: P51170 |
-Antibody , 2 types, 4 molecules DEFG
#4: Antibody | Mass: 10060.393 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Mus musculus (house mouse) / Production host: Mus musculus (house mouse) #5: Antibody | Mass: 9805.078 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Mus musculus (house mouse) / Production host: Mus musculus (house mouse) |
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-Sugars , 2 types, 7 molecules
#6: Polysaccharide | Source method: isolated from a genetically manipulated source #8: Sugar | ChemComp-NAG / |
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-Non-polymers , 1 types, 1 molecules
#7: Chemical | ChemComp-NA / |
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-Details
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: Full-length human ENaC heterotrimer bound with two high-affinity Fabs Type: COMPLEX / Entity ID: #1-#5 / Source: RECOMBINANT |
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Molecular weight | Value: 0.32 MDa / Experimental value: NO |
Source (natural) | Organism: Homo sapiens (human) |
Source (recombinant) | Organism: Homo sapiens (human) / Cell: HEK293T/17 / Plasmid: pBacMam |
Buffer solution | pH: 7.4 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Specimen support | Details: 15 mA / Grid material: GOLD / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R1.2/1.3 |
Vitrification | Instrument: FEI VITROBOT MARK III / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 285 K Details: 3.5 uL applied, manual blot, fresh 3.5 uL applied, vitrobot blot and freeze |
-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 |
Image recording | Electron dose: 60 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
-Processing
Software | Name: PHENIX / Version: 1.18.1_3865: / Classification: refinement | ||||||||||||||||||||||||
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||
3D reconstruction | Resolution: 3.06 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 252071 / Symmetry type: POINT | ||||||||||||||||||||||||
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