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- EMDB-1787: Asymmetric reconstruction of doublecortin-stabilised microtubules... -

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Basic information

Database: EMDB / ID: 1787
TitleAsymmetric reconstruction of doublecortin-stabilised microtubules decorated with kinesin motor domain
Map dataThis is a C1 reconstruction of doublecortin-stabilised microtubule decorated with kinesin motor domain
SampleMicrotubules co-polymerised with doublecortin and bound with kinesin motor domain:
Alpha-beta tubulin dimer / Doublecortin / Kinesin motor domain
KeywordsTubulin / MAP / microtubule / stabilisation / doublecortin / kinesin
SourceBos taurus (cattle) / Homo sapiens (human) / Rattus norvegicus (Norway rat)
Methodsingle particle reconstruction / cryo EM / 13.5 Å resolution
AuthorsFourniol FJ / Sindelar CV / Amigues B / Clare D / Thomas G / Perderiset M / Francis F / Houdusse A / Moores CA
CitationJournal: J. Cell Biol. / Year: 2010
Title: Template-free 13-protofilament microtubule-MAP assembly visualized at 8 A resolution.
Authors: Franck J Fourniol / Charles V Sindelar / Béatrice Amigues / Daniel K Clare / Geraint Thomas / Mylène Perderiset / Fiona Francis / Anne Houdusse / Carolyn A Moores
DateDeposition: Sep 15, 2010 / Header (metadata) release: Sep 24, 2010 / Map release: Oct 29, 2010 / Last update: Sep 19, 2012

Structure visualization

  • Surface view with section colored by density value
  • Surface level: 3
  • Imaged by UCSF Chimera
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  • Surface view colored by cylindrical radius
  • Surface level: 3
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
Supplemental images

Downloads & links


Fileemd_1787.map.gz (map file in CCP4 format, 39367 KB)
Projections & slices

Image control

AxesZ (Sec.)Y (Row.)X (Col.)
216 pix
2.8 Å/pix.
= 604.8 Å
216 pix
2.8 Å/pix.
= 604.8 Å
216 pix
2.8 Å/pix.
= 604.8 Å



Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 2.8 Å
Contour Level:3 (by author), 3 (movie #1):
Minimum - Maximum-3.84556389 - 7.45892239
Average (Standard dev.)0E-8 (1)


Space Group Number1
Map Geometry
Axis orderXYZ
CellA=B=C: 604.8 Å
α=β=γ: 90 deg.

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z2.82.82.8
M x/y/z216216216
origin x/y/z0.0000.0000.000
length x/y/z604.800604.800604.800
start NX/NY/NZ-184-184-183
MAP C/R/S123
start NC/NR/NS000
D min/max/mean-3.8467.459-0.000

Supplemental data

Sample components

Entire Microtubules co-polymerised with doublecortin and bound with kine...

EntireName: Microtubules co-polymerised with doublecortin and bound with kinesin motor domain
Number of components: 3 / Oligomeric State: 13-protofilament microtubule

Component #1: protein, Alpha-beta tubulin dimer

ProteinName: Alpha-beta tubulin dimer / a.k.a: Tubulin dimer / Oligomeric Details: Heterodimer / Recombinant expression: No
SourceSpecies: Bos taurus (cattle)
Source (natural)Location in cell: Cytoplasm / Organ or tissue: Brain

Component #2: protein, Doublecortin

ProteinName: Doublecortin / a.k.a: DCX / Oligomeric Details: Monomer / Recombinant expression: Yes
SourceSpecies: Homo sapiens (human)
Source (engineered)Expression System: Spodoptera frugiperda (fall armyworm) / Vector: pFastBac
Source (natural)Location in cell: Cytoplasm

Component #3: protein, Kinesin motor domain

ProteinName: Kinesin motor domain / a.k.a: Kinesin head / Oligomeric Details: Monomer / Recombinant expression: Yes
SourceSpecies: Rattus norvegicus (Norway rat)
Source (engineered)Expression System: Escherichia coli (E. coli)
Source (natural)Location in cell: Cytoplasm

Experimental details

Sample preparation

SpecimenSpecimen state: particle / Method: cryo EM
Sample solutionBuffer solution: 80mM Pipes, 1mM EGTA, 3mM MgCl2, 1mM TCEP, 0.5mM GTP
pH: 6.8
Support film300 mesh lacey carbon grids
VitrificationInstrument: FEI VITROBOT / Cryogen name: ETHANE / Humidity: 100 % / Method: Chamber at 37 degrees C, blot 2s / Details: Vitrification instrument: Vitrobot (FEI)

Electron microscopy imaging

Experimental equipment
Model: Tecnai F20 / Image courtesy: FEI Company
ImagingMicroscope: FEI TECNAI F20
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 200 kV / Electron dose: 15 e/Å2 / Illumination mode: FLOOD BEAM
LensMagnification: 50000 X (nominal)
Astigmatism: Objective lens astigmatism was corrected at 150,000 times magnification
Cs: 2 mm / Imaging mode: BRIGHT FIELD / Defocus: 760 - 2900 nm
Specimen HolderHolder: Eucentric / Model: OTHER / Temperature: 93 K
CameraDetector: KODAK SO-163 FILM

Image acquisition

Image acquisitionNumber of digital images: 63 / Scanner: ZEISS SCAI / Sampling size: 7 microns / Bit depth: 8 / Details: Images were binned with a factor of 2

Image processing

ProcessingMethod: single particle reconstruction
3D reconstructionAlgorithm: Custom scripts, projection matching / Software: SPIDER, FREALIGN / CTF correction: FREALIGN
Details: C1 map calculated from approximately 13,000 one-dimer-long microtubule segments
Resolution: 13.5 Å / Resolution method: FSC 0.5

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