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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-2271 | |||||||||
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Title | Cryo EM map of the Gaalphaq-PLCbeta3 complex. | |||||||||
![]() | Reconstruction of the PLCbeta3 distal CTD in contact with the PLCbeta3 catalytic core. | |||||||||
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![]() | GTP-binding protein alpha subunits / phospholipase C beta / coiled-coil domain / PH DOMAIN / EF HAND / C2 DOMAIN / TIM BARREL DOMAIN / phospholipase / GTP hydrolysis / G-protein signaling / membrane targeting / lipase / calcium binding / phospholipids / GTP-BINDING PROTEIN-HYDROLASE complex | |||||||||
Function / homology | Phosphatidylinositol-4, 5-bisphosphate phosphodiesterase beta / G-protein alpha subunit, group Q / : / G protein-coupled receptor signaling pathway![]() | |||||||||
Biological species | ![]() ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 21.0 Å | |||||||||
![]() | Dutta S / Lyon AM / Tesmer JJG / Skiniotis G | |||||||||
![]() | ![]() Title: Full-length Gα(q)-phospholipase C-β3 structure reveals interfaces of the C-terminal coiled-coil domain. Authors: Angeline M Lyon / Somnath Dutta / Cassandra A Boguth / Georgios Skiniotis / John J G Tesmer / ![]() Abstract: Phospholipase C-β (PLCβ) is directly activated by Gαq, but the molecular basis for how its distal C-terminal domain (CTD) contributes to maximal activity is poorly understood. Herein we present ...Phospholipase C-β (PLCβ) is directly activated by Gαq, but the molecular basis for how its distal C-terminal domain (CTD) contributes to maximal activity is poorly understood. Herein we present both the crystal structure and cryo-EM three-dimensional reconstructions of human full-length PLCβ3 in complex with mouse Gαq. The distal CTD forms an extended monomeric helical bundle consisting of three antiparallel segments with structural similarity to membrane-binding bin-amphiphysin-Rvs (BAR) domains. Sequence conservation of the distal CTD suggests putative membrane and protein interaction sites, the latter of which bind the N-terminal helix of Gαq in both the crystal structure and cryo-EM reconstructions. Functional analysis suggests that the distal CTD has roles in membrane targeting and in optimizing the orientation of the catalytic core at the membrane for maximal rates of lipid hydrolysis. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 953 KB | ![]() | |
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Header (meta data) | ![]() ![]() | 11.5 KB 11.5 KB | Display Display | ![]() |
Images | ![]() | 38.8 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 194.2 KB | Display | ![]() |
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Full document | ![]() | 193.3 KB | Display | |
Data in XML | ![]() | 5 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
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Annotation | Reconstruction of the PLCbeta3 distal CTD in contact with the PLCbeta3 catalytic core. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 4.48 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : Cryo EM reconstruction of the Galphaq-PLCbeta3 complex.
Entire | Name: Cryo EM reconstruction of the Galphaq-PLCbeta3 complex. |
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Components |
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-Supramolecule #1000: Cryo EM reconstruction of the Galphaq-PLCbeta3 complex.
Supramolecule | Name: Cryo EM reconstruction of the Galphaq-PLCbeta3 complex. type: sample / ID: 1000 Details: The sample was monodisperse and had a molecular weight consistent with the theoretical weight. Oligomeric state: Dimer / Number unique components: 2 |
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Molecular weight | Experimental: 1.8 MDa / Theoretical: 1.8 MDa Method: Size-exclusion chromatography and multi-angle light scattering |
-Macromolecule #1: G alpha q
Macromolecule | Name: G alpha q / type: protein_or_peptide / ID: 1 / Name.synonym: Gaq / Number of copies: 1 / Oligomeric state: monomer / Recombinant expression: Yes |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Experimental: 44 KDa / Theoretical: 44 KDa |
Recombinant expression | Organism: ![]() |
Sequence | GO: GO: 0007202 / InterPro: G-protein alpha subunit, group Q |
-Macromolecule #2: phospholipase C beta 3
Macromolecule | Name: phospholipase C beta 3 / type: protein_or_peptide / ID: 2 / Name.synonym: PLCbeta3 / Number of copies: 1 / Oligomeric state: monomer / Recombinant expression: Yes |
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Source (natural) | Organism: ![]() |
Molecular weight | Experimental: 139 KDa / Theoretical: 139 KDa |
Recombinant expression | Organism: ![]() |
Sequence | GO: G protein-coupled receptor signaling pathway InterPro: Phosphatidylinositol-4, 5-bisphosphate phosphodiesterase beta |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 1 mg/mL |
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Buffer | pH: 8 Details: 20 mM HEPES pH 8, 200 mM NaCl, 2 mM DTT, 0.9 mM CaCl2, 50 microM GDP, 30 microM AlCl3, 10 mM NaF, 5 mM MgCl2 |
Grid | Details: glow-discharged Quantifoil R2/200 mesh grid |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK IV Details: Vitrification carried out in liquid nitrogen atmosphere. Method: Blot for 2 seconds at 3 microliter sample. |
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Electron microscopy
Microscope | FEI TECNAI F20 |
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Temperature | Min: 89 K / Max: 95 K / Average: 89 K |
Alignment procedure | Legacy - Astigmatism: Objective lens astigmatism was corrected at 135,000 times magnification. |
Date | Oct 20, 2011 |
Image recording | Category: CCD / Film or detector model: GATAN ULTRASCAN 4000 (4k x 4k) / Number real images: 350 / Average electron dose: 20 e/Å2 |
Electron beam | Acceleration voltage: 120 kV / Electron source: ![]() |
Electron optics | Calibrated magnification: 71138 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2 mm / Nominal defocus max: 3.5 µm / Nominal defocus min: 1.5 µm / Nominal magnification: 50000 |
Sample stage | Specimen holder: Used two holders. / Specimen holder model: OTHER |
Experimental equipment | ![]() Model: Tecnai F20 / Image courtesy: FEI Company |
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Image processing
CTF correction | Details: each particle |
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Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 21.0 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: EMAN1 / Number images used: 12673 |
Final angle assignment | Details: EMAN1 |
-Atomic model buiding 1
Initial model | PDB ID: |
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Refinement | Space: REAL |