+Open data
-Basic information
Entry | Database: PDB / ID: 7amk | ||||||
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Title | Zebrafish RET Cadherin Like Domains 1 to 4. | ||||||
Components | Proto-oncogene tyrosine-protein kinase receptor Ret | ||||||
Keywords | SIGNALING PROTEIN / Ligand recognition / receptor tyrosine kinase / glycosylation | ||||||
Function / homology | Function and homology information branchiomeric skeletal muscle development / pronephros morphogenesis / RAF/MAP kinase cascade / : / enteric nervous system development / neural crest cell migration involved in autonomic nervous system development / axon extension / positive regulation of kinase activity / homophilic cell adhesion via plasma membrane adhesion molecules / transmembrane receptor protein tyrosine kinase activity ...branchiomeric skeletal muscle development / pronephros morphogenesis / RAF/MAP kinase cascade / : / enteric nervous system development / neural crest cell migration involved in autonomic nervous system development / axon extension / positive regulation of kinase activity / homophilic cell adhesion via plasma membrane adhesion molecules / transmembrane receptor protein tyrosine kinase activity / receptor protein-tyrosine kinase / cell surface receptor protein tyrosine kinase signaling pathway / protein-containing complex assembly / receptor complex / membrane raft / axon / calcium ion binding / protein homodimerization activity / ATP binding / plasma membrane Similarity search - Function | ||||||
Biological species | Danio rerio (zebrafish) | ||||||
Method | X-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.2 Å | ||||||
Authors | Purkiss, A.G. / McDonald, N.Q. / Goodman, K.M. / Narowtek, A. / Knowles, P.P. | ||||||
Citation | Journal: Structure / Year: 2021 Title: A two-site flexible clamp mechanism for RET-GDNF-GFRα1 assembly reveals both conformational adaptation and strict geometric spacing. Authors: Sarah E Adams / Andrew G Purkiss / Phillip P Knowles / Andrea Nans / David C Briggs / Annabel Borg / Christopher P Earl / Kerry M Goodman / Agata Nawrotek / Aaron J Borg / Pauline B McIntosh ...Authors: Sarah E Adams / Andrew G Purkiss / Phillip P Knowles / Andrea Nans / David C Briggs / Annabel Borg / Christopher P Earl / Kerry M Goodman / Agata Nawrotek / Aaron J Borg / Pauline B McIntosh / Francesca M Houghton / Svend Kjær / Neil Q McDonald / Abstract: RET receptor tyrosine kinase plays vital developmental and neuroprotective roles in metazoans. GDNF family ligands (GFLs) when bound to cognate GFRα co-receptors recognize and activate RET ...RET receptor tyrosine kinase plays vital developmental and neuroprotective roles in metazoans. GDNF family ligands (GFLs) when bound to cognate GFRα co-receptors recognize and activate RET stimulating its cytoplasmic kinase function. The principles for RET ligand-co-receptor recognition are incompletely understood. Here, we report a crystal structure of the cadherin-like module (CLD1-4) from zebrafish RET revealing interdomain flexibility between CLD2 and CLD3. Comparison with a cryo-electron microscopy structure of a ligand-engaged zebrafish RET-GDNF-GFRα1a complex indicates conformational changes within a clade-specific CLD3 loop adjacent to the co-receptor. Our observations indicate that RET is a molecular clamp with a flexible calcium-dependent arm that adapts to different GFRα co-receptors, while its rigid arm recognizes a GFL dimer to align both membrane-proximal cysteine-rich domains. We also visualize linear arrays of RET-GDNF-GFRα1a suggesting that a conserved contact stabilizes higher-order species. Our study reveals that ligand-co-receptor recognition by RET involves both receptor plasticity and strict spacing of receptor dimers by GFL ligands. | ||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 7amk.cif.gz | 495.6 KB | Display | PDBx/mmCIF format |
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PDB format | pdb7amk.ent.gz | 339.9 KB | Display | PDB format |
PDBx/mmJSON format | 7amk.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/am/7amk ftp://data.pdbj.org/pub/pdb/validation_reports/am/7amk | HTTPS FTP |
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-Related structure data
Related structure data | 7ab8C 7amlC 2x2uS C: citing same article (ref.) S: Starting model for refinement |
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Similar structure data |
-Links
-Assembly
Deposited unit |
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1 |
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Unit cell |
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Noncrystallographic symmetry (NCS) | NCS oper: (Code: givenMatrix: (0.430793954191, -0.900333557273, -0.0617742234261), (-0.902265964183, -0.431079226797, -0.00931826708919), (-0.0182400359145, 0.0597510323868, -0.998046649821)Vector: ...NCS oper: (Code: given Matrix: (0.430793954191, -0.900333557273, -0.0617742234261), Vector: |
-Components
-Protein , 1 types, 2 molecules AB
#1: Protein | Mass: 54610.270 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Danio rerio (zebrafish) / Gene: ret, ret1 / Production host: unidentified baculovirus References: UniProt: A8E7C6, receptor protein-tyrosine kinase |
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-Sugars , 7 types, 12 molecules
#2: Polysaccharide | alpha-D-mannopyranose-(1-6)-alpha-D-mannopyranose-(1-3)-[alpha-D-mannopyranose-(1-6)]beta-D- ...alpha-D-mannopyranose-(1-6)-alpha-D-mannopyranose-(1-3)-[alpha-D-mannopyranose-(1-6)]beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-[alpha-L-fucopyranose-(1-6)]2-acetamido-2-deoxy-beta-D-glucopyranose Source method: isolated from a genetically manipulated source | ||||||||||
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#3: Polysaccharide | Source method: isolated from a genetically manipulated source #4: Polysaccharide | Source method: isolated from a genetically manipulated source #5: Polysaccharide | alpha-D-mannopyranose-(1-6)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1- ...alpha-D-mannopyranose-(1-6)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-[alpha-L-fucopyranose-(1-6)]2-acetamido-2-deoxy-beta-D-glucopyranose | Source method: isolated from a genetically manipulated source #6: Polysaccharide | beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-[alpha-L-fucopyranose-(1- ...beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-[alpha-L-fucopyranose-(1-6)]2-acetamido-2-deoxy-beta-D-glucopyranose | Source method: isolated from a genetically manipulated source #7: Polysaccharide | alpha-D-mannopyranose-(1-3)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1- ...alpha-D-mannopyranose-(1-3)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-[alpha-L-fucopyranose-(1-6)]2-acetamido-2-deoxy-beta-D-glucopyranose | Source method: isolated from a genetically manipulated source #9: Sugar | |
-Non-polymers , 6 types, 245 molecules
#8: Chemical | ChemComp-CA / #10: Chemical | ChemComp-GOL / #11: Chemical | ChemComp-PGE / #12: Chemical | ChemComp-PEG / #13: Chemical | ChemComp-MES / | #14: Water | ChemComp-HOH / | |
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-Details
Has ligand of interest | N |
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-Experimental details
-Experiment
Experiment | Method: X-RAY DIFFRACTION / Number of used crystals: 1 |
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-Sample preparation
Crystal | Density Matthews: 3.28 Å3/Da / Density % sol: 62.53 % |
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Crystal grow | Temperature: 293 K / Method: vapor diffusion, sitting drop / pH: 6.2 / Details: 50 mM MES (pH 6.2), 31.5 % PEG MME 350 (v/v) |
-Data collection
Diffraction | Mean temperature: 100 K / Serial crystal experiment: N |
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Diffraction source | Source: SYNCHROTRON / Site: Diamond / Beamline: I03 / Wavelength: 0.9787 Å |
Detector | Type: DECTRIS PILATUS 6M / Detector: PIXEL / Date: May 24, 2015 / Details: Mirrors |
Radiation | Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray |
Radiation wavelength | Wavelength: 0.9787 Å / Relative weight: 1 |
Reflection | Resolution: 2.08→65.96 Å / Num. obs: 79868 / % possible obs: 98.2 % / Redundancy: 3.4 % / Biso Wilson estimate: 28.87 Å2 / CC1/2: 0.996 / Rmerge(I) obs: 0.08 / Rpim(I) all: 0.051 / Rrim(I) all: 0.095 / Net I/σ(I): 6.2 |
Reflection shell | Resolution: 2.08→2.19 Å / Redundancy: 3.3 % / Rmerge(I) obs: 1.186 / Mean I/σ(I) obs: 1.4 / Num. unique obs: 11549 / CC1/2: 0.58 / Rpim(I) all: 0.765 / Rrim(I) all: 1.416 / % possible all: 97.2 |
-Processing
Software |
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Refinement | Method to determine structure: MOLECULAR REPLACEMENT Starting model: 2x2u Resolution: 2.2→65.96 Å / SU ML: 0.3521 / Cross valid method: FREE R-VALUE / σ(F): 1.96 / Phase error: 29.5782 Stereochemistry target values: GeoStd + Monomer Library + CDL v1.2
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Solvent computation | Shrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Displacement parameters | Biso mean: 39.39 Å2 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Refinement step | Cycle: LAST / Resolution: 2.2→65.96 Å
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Refine LS restraints |
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LS refinement shell |
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Refinement TLS params. | Method: refined / Refine-ID: X-RAY DIFFRACTION
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Refinement TLS group | Refine-ID: X-RAY DIFFRACTION
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