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- EMDB-44396: Cross-linked Contactin 2 Ig1-Ig6 -

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Basic information

Entry
Database: EMDB / ID: EMD-44396
TitleCross-linked Contactin 2 Ig1-Ig6
Map data
Sample
  • Organelle or cellular component: Contactin 2 Ig1-Ig6
    • Protein or peptide: Contactin-2
  • Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose
Keywordscontactins / adhesion molecule / protein structure / conformational changes / homodimer / MEMBRANE PROTEIN
Function / homology
Function and homology information


establishment of protein localization to juxtaparanode region of axon / presynaptic membrane organization / regulation of axon diameter / positive regulation of adenosine receptor signaling pathway / L1CAM interactions / regulation of astrocyte differentiation / reduction of food intake in response to dietary excess / clustering of voltage-gated potassium channels / cerebral cortex GABAergic interneuron migration / dendrite self-avoidance ...establishment of protein localization to juxtaparanode region of axon / presynaptic membrane organization / regulation of axon diameter / positive regulation of adenosine receptor signaling pathway / L1CAM interactions / regulation of astrocyte differentiation / reduction of food intake in response to dietary excess / clustering of voltage-gated potassium channels / cerebral cortex GABAergic interneuron migration / dendrite self-avoidance / protein localization to juxtaparanode region of axon / NrCAM interactions / central nervous system myelination / cell-cell adhesion mediator activity / positive regulation of protein processing / G protein-coupled adenosine receptor signaling pathway / axon initial segment / NCAM1 interactions / juxtaparanode region of axon / node of Ranvier / regulation of cell morphogenesis / axonal fasciculation / adult walking behavior / negative regulation of neuron differentiation / fat cell differentiation / homophilic cell adhesion via plasma membrane adhesion molecules / regulation of neuronal synaptic plasticity / side of membrane / axon guidance / learning / establishment of localization in cell / synapse organization / protein processing / receptor internalization / microtubule cytoskeleton organization / myelin sheath / carbohydrate binding / postsynaptic membrane / cell adhesion / axon / neuronal cell body / synapse / cell surface / plasma membrane
Similarity search - Function
: / Immunoglobulin domain / Immunoglobulin I-set / Immunoglobulin I-set domain / Fibronectin type III domain / Fibronectin type 3 domain / Immunoglobulin subtype 2 / Immunoglobulin C-2 Type / Fibronectin type-III domain profile. / Fibronectin type III ...: / Immunoglobulin domain / Immunoglobulin I-set / Immunoglobulin I-set domain / Fibronectin type III domain / Fibronectin type 3 domain / Immunoglobulin subtype 2 / Immunoglobulin C-2 Type / Fibronectin type-III domain profile. / Fibronectin type III / Fibronectin type III superfamily / Immunoglobulin subtype / Immunoglobulin / Ig-like domain profile. / Immunoglobulin-like domain / Immunoglobulin-like domain superfamily / Immunoglobulin-like fold
Similarity search - Domain/homology
Biological speciesHomo sapiens (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.51 Å
AuthorsLiu JL / Fan SF / Ren GR / Rudenko GR
Funding support United States, 1 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of Mental Health (NIH/NIMH) United States
CitationJournal: Structure / Year: 2024
Title: Molecular mechanism of contactin 2 homophilic interaction.
Authors: Shanghua Fan / Jianfang Liu / Nicolas Chofflet / Aaron O Bailey / William K Russell / Ziqi Zhang / Hideto Takahashi / Gang Ren / Gabby Rudenko /
Abstract: Contactin 2 (CNTN2) is a cell adhesion molecule involved in axon guidance, neuronal migration, and fasciculation. The ectodomains of CNTN1-CNTN6 are composed of six Ig domains (Ig1-Ig6) and four FN ...Contactin 2 (CNTN2) is a cell adhesion molecule involved in axon guidance, neuronal migration, and fasciculation. The ectodomains of CNTN1-CNTN6 are composed of six Ig domains (Ig1-Ig6) and four FN domains. Here, we show that CNTN2 forms transient homophilic interactions (K ∼200 nM). Cryo-EM structures of full-length CNTN2 and CNTN2_Ig1-Ig6 reveal a T-shaped homodimer formed by intertwined, parallel monomers. Unexpectedly, the horseshoe-shaped Ig1-Ig4 headpieces extend their Ig2-Ig3 tips outwards on either side of the homodimer, while Ig4, Ig5, Ig6, and the FN domains form a central stalk. Cross-linking mass spectrometry and cell-based binding assays confirm the 3D assembly of the CNTN2 homodimer. The interface mediating homodimer formation differs between CNTNs, as do the homophilic versus heterophilic interaction mechanisms. The CNTN family thus encodes a versatile molecular platform that supports a very diverse portfolio of protein interactions and that can be leveraged to strategically guide neural circuit development.
History
DepositionApr 3, 2024-
Header (metadata) releaseJul 17, 2024-
Map releaseJul 17, 2024-
UpdateNov 20, 2024-
Current statusNov 20, 2024Processing site: RCSB / Status: Released

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Structure visualization

Downloads & links

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Map

FileDownload / File: emd_44396.map.gz / Format: CCP4 / Size: 125 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Voxel sizeX=Y=Z: 1.05 Å
Density
Contour LevelBy AUTHOR: 0.006
Minimum - Maximum-0.047280334 - 1.8716528
Average (Standard dev.)0.00031520185 (±0.009479568)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions320320320
Spacing320320320
CellA=B=C: 336.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Contactin 2 Ig1-Ig6

EntireName: Contactin 2 Ig1-Ig6
Components
  • Organelle or cellular component: Contactin 2 Ig1-Ig6
    • Protein or peptide: Contactin-2
  • Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose

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Supramolecule #1: Contactin 2 Ig1-Ig6

SupramoleculeName: Contactin 2 Ig1-Ig6 / type: organelle_or_cellular_component / ID: 1 / Parent: 0 / Macromolecule list: #1
Source (natural)Organism: Homo sapiens (human)

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Macromolecule #1: Contactin-2

MacromoleculeName: Contactin-2 / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 65.394629 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: ELGTGLESQT TFGPVFEDQP LSVLFPEEST EEQVLLACRA RASPPATYRW KMNGTEMKLE PGSRHQLVGG NLVIMNPTKA QDAGVYQCL ASNPVGTVVS REAILRFGFL QEFSKEERDP VKAHEGWGVM LPCNPPAHYP GLSYRWLLNE FPNFIPTDGR H FVSQTTGN ...String:
ELGTGLESQT TFGPVFEDQP LSVLFPEEST EEQVLLACRA RASPPATYRW KMNGTEMKLE PGSRHQLVGG NLVIMNPTKA QDAGVYQCL ASNPVGTVVS REAILRFGFL QEFSKEERDP VKAHEGWGVM LPCNPPAHYP GLSYRWLLNE FPNFIPTDGR H FVSQTTGN LYIARTNASD LGNYSCLATS HMDFSTKSVF SKFAQLNLAA EDTRLFAPSI KARFPAETYA LVGQQVTLEC FA FGNPVPR IKWRKVDGSL SPQWTTAEPT LQIPSVSFED EGTYECEAEN SKGRDTVQGR IIVQAQPEWL KVISDTEADI GSN LRWGCA AAGKPRPTVR WLRNGEPLAS QNRVEVLAGD LRFSKLSLED SGMYQCVAEN KHGTIYASAE LAVQALAPDF RLNP VRRLI PAARGGEILI PCQPRAAPKA VVLWSKGTEI LVNSSRVTVT PDGTLIIRNI SRSDEGKYTC FAENFMGKAN STGIL SVRD ATKITLAPSS ADINLGDNLT LQCHASHDPT MDLTFTWTLD DFPIDFDKPG GHYRRTNVKE TIGDLTILNA QLRHGG KYT CMAQTVVDSA SKEATVLVRG SASTSHHHHH H

UniProtKB: Contactin-2

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Macromolecule #2: 2-acetamido-2-deoxy-beta-D-glucopyranose

MacromoleculeName: 2-acetamido-2-deoxy-beta-D-glucopyranose / type: ligand / ID: 2 / Number of copies: 10 / Formula: NAG
Molecular weightTheoretical: 221.208 Da
Chemical component information

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

Concentration0.025 mg/mL
BufferpH: 8
Component:
ConcentrationNameFormula
20.0 mMHEPES
50.0 mMSodium chlorideNaCl

Details: 20 mM HEPES pH 8.0, 50 mM NaCl
VitrificationCryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 281 K / Instrument: LEICA EM GP

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Electron microscopy

MicroscopeTFS KRIOS
Specialist opticsEnergy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 1 / Number real images: 4297 / Average exposure time: 7.39 sec. / Average electron dose: 50.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 1.6 µm / Nominal defocus min: 0.6 µm / Nominal magnification: 81000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: NONE
Details: The initial volume was generated by ab initio reconstruction.
Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.51 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 4.3.0) / Number images used: 98082
Initial angle assignmentType: RANDOM ASSIGNMENT / Software - Name: cryoSPARC (ver. 4.3.0)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 4.3.0)

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Atomic model buiding 1

RefinementProtocol: FLEXIBLE FIT
Output model

PDB-9ba5:
Cross-linked Contactin 2 Ig1-Ig6

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