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- EMDB-11306: Cryo-STET tomography of T. brucei FAZ filament -

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Basic information

Entry
Database: EMDB / ID: EMD-11306
TitleCryo-STET tomography of T. brucei FAZ filament
Map dataCryo-STET reconstruction of the anterior end of a T. brucei bloodstream cell.
Sample
  • Cell: FAZ filament T. brucei
Biological speciesTrypanosoma brucei brucei (eukaryote)
Methodelectron tomography / cryo EM
AuthorsTrepout S
Funding support France, 2 items
OrganizationGrant numberCountry
French National Research AgencyANR-11-BSV8-016 France
French National Research AgencyANR-15-CE11-0002 France
CitationJournal: J Struct Biol X / Year: 2020
Title: structural analysis of the flagellum attachment zone in using cryo-scanning transmission electron tomography.
Authors: Sylvain Trépout /
Abstract: The flagellum of is a 20 µm-long organelle responsible for locomotion and cell morphogenesis. The flagellum attachment zone (FAZ) is a multi-protein complex whose function is to attach the ...The flagellum of is a 20 µm-long organelle responsible for locomotion and cell morphogenesis. The flagellum attachment zone (FAZ) is a multi-protein complex whose function is to attach the flagellum to the cell body but also to guide cytokinesis. Cryo-transmission electron microscopy is a tool of choice to access the structure of the FAZ in a close-to-native state. However, because of the large dimension of the cell body, the whole FAZ cannot be structurally studied at the nanometre scale in 3D using classical transmission electron microscopy approaches. In the present work, cryo-scanning transmission electron tomography, a new method capable of investigating cryo-fixed thick biological samples, has been used to study whole cells at the bloodstream stage. The method has been used to visualise and characterise the structure and organisation of the FAZ filament. It is composed of an array of cytoplasmic stick-like structures. These sticks are heterogeneously distributed between the posterior part and the anterior tip of the cell. This cryo-STET investigation provides new insights into the structure of the FAZ filament. In combination with protein structure predictions, this work proposes a new model for the elongation of the FAZ.
History
DepositionJul 5, 2020-
Header (metadata) releaseJul 15, 2020-
Map releaseJul 15, 2020-
UpdateJul 28, 2021-
Current statusJul 28, 2021Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Solid view (volume rendering)
  • Imaged by UCSF Chimera
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  • Solid view (volume rendering)
  • Imaged by UCSF Chimera
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Supplemental images

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Map

FileDownload / File: emd_11306.map.gz / Format: CCP4 / Size: 259 MB / Type: IMAGE STORED AS SIGNED INTEGER (2 BYTES)
AnnotationCryo-STET reconstruction of the anterior end of a T. brucei bloodstream cell.
Voxel sizeX=Y=Z: 32 Å
Density
Minimum - Maximum0.0 - 30457.0
Average (Standard dev.)20725.426 (±582.2697)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions750425426
Spacing425750426
CellA: 13600.0 Å / B: 24000.0 Å / C: 13632.0 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Integer*27
Å/pix. X/Y/Z323232
M x/y/z425750426
origin x/y/z0.0000.0000.000
length x/y/z13600.00024000.00013632.000
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS425750426
D min/max/mean0.00030457.00020725.426

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Supplemental data

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Sample components

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Entire : FAZ filament T. brucei

EntireName: FAZ filament T. brucei
Components
  • Cell: FAZ filament T. brucei

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Supramolecule #1: FAZ filament T. brucei

SupramoleculeName: FAZ filament T. brucei / type: cell / ID: 1 / Parent: 0
Source (natural)Organism: Trypanosoma brucei brucei (eukaryote)

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Experimental details

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Structure determination

Methodcryo EM
Processingelectron tomography
Aggregation statecell

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Sample preparation

BufferpH: 7.4
VitrificationCryogen name: ETHANE
SectioningOther: NO SECTIONING
Fiducial markerManufacturer: Aurion / Diameter: 15 nm

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Electron microscopy

MicroscopeJEOL 2200FS
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: OTHER / Imaging mode: BRIGHT FIELDBright-field microscopy
Sample stageSpecimen holder model: GATAN 914 HIGH TILT LIQUID NITROGEN CRYO TRANSFER TOMOGRAPHY HOLDER
Image recordingFilm or detector model: OTHER / Average electron dose: 80.0 e/Å2

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Image processing

Final reconstructionNumber images used: 72

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