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- EMDB-18651: GDNF/GFRa1 cell adhesion complex bridging between adhering liposomes. -
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Open data
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Basic information
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Title | GDNF/GFRa1 cell adhesion complex bridging between adhering liposomes. | |||||||||
![]() | Reconstructed tomogram of zGDNF-zGFRa1 complexes bridging between adhering liposomes. Tomogram reconstructions were generated with a SIRT-like filter with 5 iterations and binned by 4. | |||||||||
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![]() | Synaptic / Adhesion / Complex / LiCAM / CELL ADHESION | |||||||||
Function / homology | ![]() diencephalon development / positive regulation of ureteric bud formation / postganglionic parasympathetic fiber development / positive regulation of monooxygenase activity / regulation of dopaminergic neuron differentiation / glial cell-derived neurotrophic factor receptor binding / glial cell-derived neurotrophic factor receptor signaling pathway / regulation of morphogenesis of a branching structure / regulation of dopamine uptake involved in synaptic transmission / enteric nervous system development ...diencephalon development / positive regulation of ureteric bud formation / postganglionic parasympathetic fiber development / positive regulation of monooxygenase activity / regulation of dopaminergic neuron differentiation / glial cell-derived neurotrophic factor receptor binding / glial cell-derived neurotrophic factor receptor signaling pathway / regulation of morphogenesis of a branching structure / regulation of dopamine uptake involved in synaptic transmission / enteric nervous system development / peristalsis / positive regulation of branching involved in ureteric bud morphogenesis / sympathetic nervous system development / peripheral nervous system development / metanephros development / mRNA stabilization / neural crest cell migration / MAP kinase kinase kinase activity / branching involved in ureteric bud morphogenesis / growth factor activity / receptor tyrosine kinase binding / neuron projection development / signaling receptor activity / nervous system development / protein-containing complex assembly / negative regulation of neuron apoptotic process / receptor complex / external side of plasma membrane / positive regulation of transcription by RNA polymerase II / protein homodimerization activity / extracellular space / extracellular region Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | electron tomography / cryo EM | |||||||||
![]() | Houghton FM / Briggs DC / McDonald NQ | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Architecture and regulation of a GDNF-GFRα1 synaptic adhesion assembly. Authors: F M Houghton / S E Adams / A S Ríos / L Masino / A G Purkiss / D C Briggs / F Ledda / N Q McDonald / ![]() ![]() Abstract: Glial-cell line derived neurotrophic factor (GDNF) bound to its co-receptor GFRα1 stimulates the RET receptor tyrosine kinase, promoting neuronal survival and neuroprotection. The GDNF-GFRα1 ...Glial-cell line derived neurotrophic factor (GDNF) bound to its co-receptor GFRα1 stimulates the RET receptor tyrosine kinase, promoting neuronal survival and neuroprotection. The GDNF-GFRα1 complex also supports synaptic cell adhesion independently of RET. Here, we describe the structure of a decameric GDNF-GFRα1 assembly determined by crystallography and electron microscopy, revealing two GFRα1 pentamers bridged by five GDNF dimers. We reconsitituted the assembly between adhering liposomes and used cryo-electron tomography to visualize how the complex fulfils its membrane adhesion function. The GFRα1:GFRα1 pentameric interface was further validated both in vitro by native PAGE and in cellulo by cell-clustering and dendritic spine assays. Finally, we provide biochemical and cell-based evidence that RET and heparan sulfate cooperate to prevent assembly of the adhesion complex by competing for the adhesion interface. Our results provide a mechanistic framework to understand GDNF-driven cell adhesion, its relationship to trophic signalling, and the central role played by GFRα1. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 10.3 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 13.1 KB 13.1 KB | Display Display | ![]() |
Images | ![]() | 181.6 KB | ||
Filedesc metadata | ![]() | 5.1 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8os6C C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
File | ![]() | ||||||||||||||||||||
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Annotation | Reconstructed tomogram of zGDNF-zGFRa1 complexes bridging between adhering liposomes. Tomogram reconstructions were generated with a SIRT-like filter with 5 iterations and binned by 4. | ||||||||||||||||||||
Voxel size | X=Y=Z: 13.02 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
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Sample components
-Entire : GDNF/GFRa1 cell adhesion complex assembled onto liposomes.
Entire | Name: GDNF/GFRa1 cell adhesion complex assembled onto liposomes. |
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Components |
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-Supramolecule #1: GDNF/GFRa1 cell adhesion complex assembled onto liposomes.
Supramolecule | Name: GDNF/GFRa1 cell adhesion complex assembled onto liposomes. type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 700 KDa |
-Macromolecule #1: GDNF family receptor alpha
Macromolecule | Name: GDNF family receptor alpha / type: protein_or_peptide / ID: 1 Details: Vector-derived sequences and tag present at N- and C-terminus. Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Sequence | String: APLAEPALAK E NNCLNAAK AC NLNDTCK KYR SAYISP CTSR VSTAE VCNKR KCHK ALRQFF DKV PPKHSYG ML YCSCPLGD Q SACSERRRQ TIVPACSYED KERPNCLTL Q VSCKTNYI CR SRLADFF TNC QPEPLS LSGC LKENY ADCLL SYSG ...String: APLAEPALAK E NNCLNAAK AC NLNDTCK KYR SAYISP CTSR VSTAE VCNKR KCHK ALRQFF DKV PPKHSYG ML YCSCPLGD Q SACSERRRQ TIVPACSYED KERPNCLTL Q VSCKTNYI CR SRLADFF TNC QPEPLS LSGC LKENY ADCLL SYSG LIGTVM TPN YLRSPKI SV SPFCDCSS S GNSKEECDR FTEFFTDNAC LRNAIQAFG N GTDVSVWH PM PPVQTTT S AAAHHHHH H UniProtKB: GDNF family receptor alpha |
-Macromolecule #2: Glial cell line-derived neurotrophic factor
Macromolecule | Name: Glial cell line-derived neurotrophic factor / type: protein_or_peptide / ID: 2 / Details: Vector-derived sequence at N-terminus / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Recombinant expression | Organism: ![]() |
Sequence | String: APLA RVKGQ GR GCLLKEI HLN VTDLDL GYRT KEELI FRYCS GPCH DAETNY DKI LNNLTHN KK LDKDTPSR T CCRPIAFDD DISFLDDSLE YHTLKKHSA K KCACV UniProtKB: Glial cell line-derived neurotrophic factor |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | electron tomography |
Aggregation state | particle |
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Sample preparation
Concentration | 0.133 mg/mL | |||||||||
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Buffer | pH: 7.5 Component:
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Grid | Model: Quantifoil R2/2 / Material: COPPER / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 45 sec. / Pretreatment - Atmosphere: AIR | |||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 85 % / Chamber temperature: 295 K / Instrument: FEI VITROBOT MARK IV / Details: 3 s blot time. | |||||||||
Details | Uncrosslinked GDNF/GFRa1 complex assembled onto liposome surfaces. | |||||||||
Sectioning | Other: NO SECTIONING | |||||||||
Fiducial marker | Manufacturer: BBI Solutions / Diameter: 10 nm |
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Electron microscopy
Microscope | FEI TALOS ARCTICA |
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Image recording | Film or detector model: FEI FALCON III (4k x 4k) / Detector mode: COUNTING / Number grids imaged: 1 / Number real images: 38 / Average exposure time: 0.72 sec. / Average electron dose: 1.99 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 7.0 µm / Nominal defocus min: 4.0 µm / Nominal magnification: 45000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Talos Arctica / Image courtesy: FEI Company |
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Image processing
Final reconstruction | Algorithm: BACK PROJECTION / Software - Name: ![]() Details: The final alignment was binned by 4 and the final tomogram was generated using a back-projection algorithm with a SIRT-like filter equivalent to 5 iterations of the SIRT algorithm. Number images used: 38 |
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