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- PDB-9d77: Crystal form of Netrin-1 mimics nanotubes -

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Basic information

Entry
Database: PDB / ID: 9d77
TitleCrystal form of Netrin-1 mimics nanotubes
ComponentsNetrin-1
KeywordsSIGNALING PROTEIN / APOPTOSIS / Axon guidance cue / nanotubes
Function / homology
Function and homology information


regulation of glial cell migration / chemorepulsion of axon / anterior/posterior axon guidance / Cdc42 protein signal transduction / motor neuron migration / tissue development / negative regulation of axon extension / substrate-dependent cell migration, cell extension / motor neuron axon guidance / nuclear migration ...regulation of glial cell migration / chemorepulsion of axon / anterior/posterior axon guidance / Cdc42 protein signal transduction / motor neuron migration / tissue development / negative regulation of axon extension / substrate-dependent cell migration, cell extension / motor neuron axon guidance / nuclear migration / positive regulation of cell motility / inner ear morphogenesis / regulation of synapse assembly / dendrite development / basement membrane / positive regulation of axon extension / glial cell proliferation / positive regulation of glial cell proliferation / cell periphery / animal organ morphogenesis / cell-cell adhesion / actin cytoskeleton / Ras protein signal transduction / extracellular region / nucleoplasm / cytosol
Similarity search - Function
Laminin, N-terminal / : / Laminin N-terminal (Domain VI) / Laminin N-terminal domain profile. / Laminin N-terminal domain (domain VI) / Laminin-type EGF-like (LE) domain profile. / Laminin-type EGF-like (LE) domain signature. / Laminin-type epidermal growth factor-like domai / Laminin EGF domain / Laminin-type EGF domain ...Laminin, N-terminal / : / Laminin N-terminal (Domain VI) / Laminin N-terminal domain profile. / Laminin N-terminal domain (domain VI) / Laminin-type EGF-like (LE) domain profile. / Laminin-type EGF-like (LE) domain signature. / Laminin-type epidermal growth factor-like domai / Laminin EGF domain / Laminin-type EGF domain / Netrin C-terminal Domain / Netrin module, non-TIMP type / UNC-6/NTR/C345C module / Netrin domain / NTR domain profile. / Tissue inhibitor of metalloproteinases-like, OB-fold / EGF-like domain signature 1. / Galactose-binding-like domain superfamily
Similarity search - Domain/homology
Biological speciesGallus gallus (chicken)
MethodX-RAY DIFFRACTION / MOLECULAR REPLACEMENT / Resolution: 3.4 Å
AuthorsMeier, M. / Krahn, N.J. / McDougall, M.D. / Rafiei, F. / Koch, M. / Stetefeld, J.
Funding support Canada, Germany, 2items
OrganizationGrant numberCountry
Canadian Institutes of Health Research (CIHR)CIHR-PJT 152935 Canada
German Research Foundation (DFG)DFG-FOR2722 Germany
CitationJournal: To Be Published
Title: Mechanistic insights in the higher-order protein assemblies of netrin-1
Authors: Rafiei, F. / Meier, M. / Koch, M. / Stetefeld, J.
History
DepositionAug 16, 2024Deposition site: RCSB / Processing site: RCSB
Revision 1.0Oct 9, 2024Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Netrin-1
hetero molecules


Theoretical massNumber of molelcules
Total (without water)52,9929
Polymers49,5801
Non-polymers3,4128
Water36020
1
A: Netrin-1
hetero molecules

A: Netrin-1
hetero molecules


  • defined by author&software
  • Evidence: SAXS, Experimental evidence for monomer-dimer equilibrium by SEC-SAXS, SEC-MALS and sedimentation velocity experiments.
  • 106 kDa, 2 polymers
Theoretical massNumber of molelcules
Total (without water)105,98318
Polymers99,1602
Non-polymers6,82316
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
crystal symmetry operation4_655-x+1,-y,z1
Buried area10680 Å2
ΔGint20 kcal/mol
Surface area48320 Å2
MethodPISA
Unit cell
Length a, b, c (Å)189.260, 189.260, 45.880
Angle α, β, γ (deg.)90.000, 90.000, 120.000
Int Tables number168
Space group name H-MP6

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Components

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Protein , 1 types, 1 molecules A

#1: Protein Netrin-1


Mass: 49579.926 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Gallus gallus (chicken) / Gene: NTN1 / Cell (production host): Embryonic kidney cell / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / Tissue (production host): Embryonic kidney / References: UniProt: Q90922

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Sugars , 3 types, 4 molecules

#2: Polysaccharide alpha-D-mannopyranose-(1-3)-[alpha-D-mannopyranose-(1-6)]beta-D-mannopyranose-(1-4)-2-acetamido-2- ...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


Type: oligosaccharide / Mass: 1056.964 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
DescriptorTypeProgram
DManpa1-3[DManpa1-6]DManpb1-4DGlcpNAcb1-4[LFucpa1-6]DGlcpNAcb1-ROHGlycam Condensed SequenceGMML 1.0
WURCS=2.0/4,6,5/[a2122h-1b_1-5_2*NCC/3=O][a1122h-1b_1-5][a1122h-1a_1-5][a1221m-1a_1-5]/1-1-2-3-3-4/a4-b1_a6-f1_b4-c1_c3-d1_c6-e1WURCSPDB2Glycan 1.1.0
[][D-1-deoxy-GlcpNAc]{[(4+1)][b-D-GlcpNAc]{[(4+1)][b-D-Manp]{[(3+1)][a-D-Manp]{}[(6+1)][a-D-Manp]{}}}[(6+1)][a-L-Fucp]{}}LINUCSPDB-CARE
#3: Polysaccharide alpha-D-mannopyranose-(1-3)-[alpha-D-mannopyranose-(1-6)]beta-D-mannopyranose-(1-4)-2-acetamido-2- ...alpha-D-mannopyranose-(1-3)-[alpha-D-mannopyranose-(1-6)]beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose


Type: oligosaccharide / Mass: 910.823 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
DescriptorTypeProgram
DManpa1-3[DManpa1-6]DManpb1-4DGlcpNAcb1-4DGlcpNAcb1-ROHGlycam Condensed SequenceGMML 1.0
WURCS=2.0/3,5,4/[a2122h-1b_1-5_2*NCC/3=O][a1122h-1b_1-5][a1122h-1a_1-5]/1-1-2-3-3/a4-b1_b4-c1_c3-d1_c6-e1WURCSPDB2Glycan 1.1.0
[][D-1-deoxy-GlcpNAc]{[(4+1)][b-D-GlcpNAc]{[(4+1)][b-D-Manp]{[(3+1)][a-D-Manp]{}[(6+1)][a-D-Manp]{}}}}LINUCSPDB-CARE
#4: Sugar ChemComp-NAG / 2-acetamido-2-deoxy-beta-D-glucopyranose / N-acetyl-beta-D-glucosamine / 2-acetamido-2-deoxy-beta-D-glucose / 2-acetamido-2-deoxy-D-glucose / 2-acetamido-2-deoxy-glucose / N-ACETYL-D-GLUCOSAMINE


Type: D-saccharide, beta linking / Mass: 221.208 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Formula: C8H15NO6
IdentifierTypeProgram
DGlcpNAcbCONDENSED IUPAC CARBOHYDRATE SYMBOLGMML 1.0
N-acetyl-b-D-glucopyranosamineCOMMON NAMEGMML 1.0
b-D-GlcpNAcIUPAC CARBOHYDRATE SYMBOLPDB-CARE 1.0
GlcNAcSNFG CARBOHYDRATE SYMBOLGMML 1.0

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Non-polymers , 4 types, 24 molecules

#5: Chemical ChemComp-CA / CALCIUM ION


Mass: 40.078 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: Ca
#6: Chemical ChemComp-CL / CHLORIDE ION


Mass: 35.453 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: Cl
#7: Chemical ChemComp-MPD / (4S)-2-METHYL-2,4-PENTANEDIOL


Mass: 118.174 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C6H14O2 / Comment: precipitant*YM
#8: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 20 / Source method: isolated from a natural source / Formula: H2O

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Details

Has ligand of interestN
Has protein modificationY

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Experimental details

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Experiment

ExperimentMethod: X-RAY DIFFRACTION / Number of used crystals: 1

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Sample preparation

CrystalDensity Matthews: 4.53 Å3/Da / Density % sol: 72.86 %
Crystal growTemperature: 293 K / Method: vapor diffusion, hanging drop / pH: 6.5
Details: 40% v/v 2-methyl-2,4-pentanediol; 5% w/v polyethylene glycol 8000; 100 mM 2-(N-morpholino)ethanesulfonic acid; pH 6.5
Temp details: Crystallisation cabinet

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Data collection

DiffractionMean temperature: 100 K / Ambient temp details: X-stream 2000 / Serial crystal experiment: N
Diffraction sourceSource: ROTATING ANODE / Type: RIGAKU MICROMAX-007 HF / Wavelength: 1.54187 Å
DetectorType: RIGAKU RAXIS IV++ / Detector: IMAGE PLATE / Date: Jun 26, 2017 / Details: Rigaku Osmic Confocal Max-Flux multilayer mirror
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1.54187 Å / Relative weight: 1
ReflectionResolution: 3.4→35.77 Å / Num. obs: 9185 / % possible obs: 68.9 % / Redundancy: 9.6 % / Biso Wilson estimate: 83.7 Å2 / CC1/2: 0.994 / CC star: 0.999 / Rmerge(I) obs: 0.412 / Rpim(I) all: 0.14 / Rrim(I) all: 0.435 / Net I/av σ(I): 5.6 / Net I/σ(I): 4.9
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. unique obsCC1/2CC starRpim(I) allRrim(I) all% possible all
4.2-35.779.60.3385.871110.9950.9990.1160.35799.51
4.2-4.59.81.1662.413120.670.8960.3931.232100
3.4-4.29.81.4731.920740.5830.7290.4881.55333.53

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Processing

Software
NameVersionClassification
REFMAC5.8.0430refinement
HKL-3000data collection
XDSJun 30, 2023 BUILT=20230630data reduction
XSCALEJun 30, 2023 BUILT=20230630data scaling
PHASER2.8.3phasing
Coot0.9.8.95model building
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 3.4→35.77 Å / Cor.coef. Fo:Fc: 0.857 / Cor.coef. Fo:Fc free: 0.805 / SU B: 170.905 / SU ML: 1.034 / Cross valid method: THROUGHOUT / ESU R Free: 0.864
Details: Hydrogens have been added in their riding positions
RfactorNum. reflection% reflectionSelection details
Rfree0.3348 743 8.089 %Random selection
Rwork0.2967 8442 --
all0.3 ---
obs-9185 68.74 %-
Solvent computationIon probe radii: 0.7 Å / Shrinkage radii: 0.7 Å / VDW probe radii: 0.85 Å / Solvent model: MASK BULK SOLVENT
Displacement parametersBiso mean: 88.104 Å2
Baniso -1Baniso -2Baniso -3
1--4.198 Å2-2.099 Å2-0 Å2
2---4.198 Å20 Å2
3---13.618 Å2
Refinement stepCycle: LAST / Resolution: 3.4→35.77 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms3272 0 225 20 3517
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0070.0123679
X-RAY DIFFRACTIONr_bond_other_d0.0010.0163254
X-RAY DIFFRACTIONr_angle_refined_deg1.8471.8955023
X-RAY DIFFRACTIONr_angle_other_deg0.61.8457565
X-RAY DIFFRACTIONr_dihedral_angle_1_deg10.3524.693483
X-RAY DIFFRACTIONr_dihedral_angle_other_1_deg8.932108
X-RAY DIFFRACTIONr_dihedral_angle_2_deg7.406535
X-RAY DIFFRACTIONr_dihedral_angle_3_deg17.9499.98601
X-RAY DIFFRACTIONr_dihedral_angle_other_3_deg0.47841
X-RAY DIFFRACTIONr_dihedral_angle_6_deg14.04210170
X-RAY DIFFRACTIONr_chiral_restr0.0570.2570
X-RAY DIFFRACTIONr_gen_planes_refined0.0070.024266
X-RAY DIFFRACTIONr_gen_planes_other0.0020.02863
X-RAY DIFFRACTIONr_nbd_refined0.1810.2674
X-RAY DIFFRACTIONr_symmetry_nbd_other0.1950.23184
X-RAY DIFFRACTIONr_nbtor_refined0.1740.21739
X-RAY DIFFRACTIONr_symmetry_nbtor_other0.1070.21950
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.2810.2114
X-RAY DIFFRACTIONr_symmetry_xyhbond_nbd_other0.1620.24
X-RAY DIFFRACTIONr_symmetry_nbd_refined0.1050.228
X-RAY DIFFRACTIONr_nbd_other0.1620.2107
X-RAY DIFFRACTIONr_symmetry_xyhbond_nbd_refined0.1820.25
X-RAY DIFFRACTIONr_mcbond_it0.3380.4691697
X-RAY DIFFRACTIONr_mcbond_other0.3380.4691697
X-RAY DIFFRACTIONr_mcangle_it0.610.8392124
X-RAY DIFFRACTIONr_mcangle_other0.610.8392125
X-RAY DIFFRACTIONr_scbond_it0.8640.9581982
X-RAY DIFFRACTIONr_scbond_other0.8640.9571983
X-RAY DIFFRACTIONr_scangle_it1.5321.7792902
X-RAY DIFFRACTIONr_scangle_other1.5321.7792903
X-RAY DIFFRACTIONr_lrange_it2.8537.41414412
X-RAY DIFFRACTIONr_lrange_other2.8537.41414412
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
3.4-3.5820.35120.412281X-RAY DIFFRACTION15.3403
3.582-3.7970.371500.347454X-RAY DIFFRACTION27.8607
3.797-4.0560.378550.337692X-RAY DIFFRACTION43.761
4.056-4.3760.3751190.3651209X-RAY DIFFRACTION84.6939
4.376-4.7870.3391180.3391336X-RAY DIFFRACTION99.1814
4.787-5.340.36970.3081236X-RAY DIFFRACTION99.4034
5.34-6.1430.442980.311091X-RAY DIFFRACTION99.5812
6.143-7.4690.265820.26942X-RAY DIFFRACTION99.8051
7.469-10.340.278630.212744X-RAY DIFFRACTION99.6296
10.34-35.770.268490.209457X-RAY DIFFRACTION98.6355
Refinement TLS params.

Method: refined / Refine-ID: X-RAY DIFFRACTION

IDL112)L122)L132)L222)L232)L332)S11 (Å °)S12 (Å °)S13 (Å °)S21 (Å °)S22 (Å °)S23 (Å °)S31 (Å °)S32 (Å °)S33 (Å °)T112)T122)T132)T222)T232)T332)Origin x (Å)Origin y (Å)Origin z (Å)
13.47492.68832.42875.20022.97213.86750.3762-0.50520.61250.5017-0.58760.46470.0391-1.00330.21131.0661-0.30910.04361.37770.00330.136443.11-50.852-26.957
24.80582.0352-0.79982.21510.79451.08970.3981-0.23260.74290.17-0.83260.9449-0.1637-0.60920.43451.81-0.1953-0.16581.2805-0.2580.451657.197-26.111-16.432
38.5333-1.02150.70640.1246-0.11231.62010.02050.7761-0.3155-0.0492-0.09410.04150.03-0.17870.07361.3255-0.0713-0.12430.6116-0.09910.037376.503-10.088-10.754
47.98650.0523-1.49830.0286-0.26813.030.1051-0.42390.9289-0.03490.03730.0026-0.3701-0.2348-0.14241.27580.05030.01860.7128-0.04870.108596.86414.224-5.994
51.6440.7734-6.67441.0539-4.70730.83330.5743-0.18310.1292-0.2544-0.0812-0.0065-0.9181.1163-0.49311.32040.0246-0.6611.31230.07441.021921.164-44.552-56.182
62.37-1.41791.878511.1903-14.345918.3979-0.7904-0.6456-0.4724-0.4649-0.2996-0.82970.53330.34421.08991.1061-0.02430.12070.8359-0.21120.683257.75-53.157-42.518
77.77-5.9445-9.89864.56167.578212.6125-0.58290.5578-0.6950.4575-0.29990.54830.7467-0.6650.88280.8204-0.2545-0.28931.35920.32840.614294.382-6.0334.948
8773.9279-373.2213-112.5057180.171454.128516.44042.31834.93664.9768-1.024-1.8452-2.7304-0.3719-1.0669-0.47311.2936-0.11330.18571.5418-0.94520.709780.55-31.034-13.896
911.29866.8435-31.18714.1512-18.894986.08981.69160.01130.5841.0429-0.0710.3591-4.72730.0248-1.62061.0317-0.11220.07280.8888-0.08210.0663103.1067.58-4.104
Refinement TLS group
IDRefine-IDRefine TLS-IDSelectionAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1ALLA37 - 286
2X-RAY DIFFRACTION1ALLA458
3X-RAY DIFFRACTION1ALLW1 - 20
4X-RAY DIFFRACTION1ALLA37 - 286
5X-RAY DIFFRACTION1ALLA458
6X-RAY DIFFRACTION1ALLW1 - 20
7X-RAY DIFFRACTION2ALLA287 - 341
8X-RAY DIFFRACTION2ALLW21 - 22
9X-RAY DIFFRACTION2ALLA287 - 341
10X-RAY DIFFRACTION2ALLW21 - 22
11X-RAY DIFFRACTION3ALLA342 - 404
12X-RAY DIFFRACTION3ALLC1
13X-RAY DIFFRACTION3ALLW23
14X-RAY DIFFRACTION3ALLA342 - 404
15X-RAY DIFFRACTION3ALLC1
16X-RAY DIFFRACTION3ALLW23
17X-RAY DIFFRACTION4ALLA405 - 456
18X-RAY DIFFRACTION4ALLW24 - 25
19X-RAY DIFFRACTION4ALLA405 - 456
20X-RAY DIFFRACTION4ALLW24 - 25
21X-RAY DIFFRACTION5ALLA470 - 474
22X-RAY DIFFRACTION5ALLA470 - 474
23X-RAY DIFFRACTION6ALLA459 - 464
24X-RAY DIFFRACTION6ALLA459 - 464
25X-RAY DIFFRACTION7ALLA465 - 469
26X-RAY DIFFRACTION7ALLA465 - 469
27X-RAY DIFFRACTION8ALLL2
28X-RAY DIFFRACTION8ALLL2
29X-RAY DIFFRACTION9ALLL3
30X-RAY DIFFRACTION9ALLL3

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