EMD-7523

Helical reconstruction
4.8 Å
EMD-7523
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Deposition: 06/03/2018
Map released: 16/01/2019
Last modified: 15/05/2019
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Overview 3D View Sample Experiment Validation Volume Browser Additional data Links

EMD-7523
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EMICSS annotations (xml)

cryo-EM reconstruction of microtubule-bound 4R-tau

EMD-7523

Helical reconstruction
4.8 Å
EMD-7523
3D View Gallery
Deposition: 06/03/2018
Map released: 16/01/2019
Last modified: 15/05/2019
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links
Method: Helical reconstruction
Aggregation State: Filament
Specimen preparation [1]
Concentration: 0.5 mg/mL
Details: Tubulin concentration is 0.5 mg/mL, tau concentration is 1 mg/mL
Buffer
pH: 6.8
Buffer components [5]:
Name Formula Concentration ChEBI
PIPES C8H18N2O6S2 80.0 mM
EGTA C14H24N2O10 1.0 mM
Magnesium Chloride MgCl2 1.0 mM
DTT C4H10O2S2 1.0 mM
Guanine tri-phosphate GTP 1.0 mM
Grid
Mesh: 400
Model: C-flat-1.2/1.3 4C
Material: COPPER
Details: Unspecified
Pretreatment
Type: GLOW DISCHARGE
Atmosphere: AIR
Support Film [1]
Material Topology Thickness
CARBON HOLEY -
Vitrification
Cryogen name: ETHANE
Chamber humidity: 100%
Chamber temperature: 310.15 K
Instrument: FEI VITROBOT MARK IV
Details: Blot force 10 pN, 6 second blot time. used C-flat 1.2/1.3 holey grids. First 2 uL of microtubules were adhered to grid for 30 seconds, followed by 2 4 uL washes with 30 second incubation for each wash..
Microscopy [1]
Microscope: FEI TITAN
Illumination mode: FLOOD BEAM
Imaging mode: BRIGHT FIELD
Electron source: FIELD EMISSION GUN
Acceleration voltage: 300 kV
C2 aperture diameter: 100.0 µm
Nominal CS: 2.7 mm
Nominal defocus: 1.0 µm - 2.5 µm
Nominal magnification: 27500.0
Specimen holder model: GATAN 626 SINGLE TILT LIQUID NITROGEN CRYO TRANSFER HOLDER
Cooling holder cryogen: NITROGEN
Alignment procedure: COMA FREE
Image Recording [1]
Detector model: GATAN K2 SUMMIT (4k x 4k)
Detector mode: COUNTING
Number of grids: 1
Number of real images: 422
Average exposure time: 6.0 s
Average electron dose per image: 27.5 e/Å2
Image processing [1]
Details: Used motioncorr to correct for beam-induced motion
Final reconstruction
Resolution: 4.8 Å ( BY AUTHOR)
Resolution method: FSC 0.143 CUT-OFF
Number of images used: 24618
Algorithm: FOURIER SPACE
Details:
⌯ Applied Symmetry
ΔΖ: 8.6 Å
ΔΦ: -25.76°
Axial symmetry: C1
Software [1]
Name Version Details
FREALIGN 9.09 -
Startup model [1]
Type: EMDB MAP
EMDB model: EMD-5194
Details:Used a naked microtubule reconstruction as the initial model, low pass filtered to 20 Angstrom
⦩ Final angle assignment
Type: NOT APPLICABLE
Software [1]
Name Version Details
FREALIGN 9.09 -
CTF correction
Software [1]
Name Version Details
CTFFIND 4 -
Segment selection [1]
Number selected Segment length Segment overlap Total filament length Details
36777 - - - manually selected helical segments, then filled in with overlapping boxes spaced 80 Angstrom apart.
Map
Format: CCP4
Data type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotation details: Cryo-EM reconstruction of shortened tau construct (residues 197-400 of full-length tau)
⬡ Geometry
X Y Z
Dimensions 512 512 512
Origin 0 0 0
Spacing 512 512 512
Voxel size 1.32 Å 1.32 Å 1.32 Å
Contour list
Primary Level Source
True 4.1 AUTHOR