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- EMDB-17769: Subtomogram average of the Campylobacter jejuni motor with basal ... -

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Basic information

Entry
Database: EMDB / ID: EMD-17769
TitleSubtomogram average of the Campylobacter jejuni motor with basal disk genes, flgPQ, expressed at very low level
Map dataCampylobacter jejuni motor with basal disk genes, flgPQ, expressed at very low level
Sample
  • Organelle or cellular component: Campylobacter flagellar motor
Keywordsflagellum / motor / Campylobacter jejuni / basal disk / MOTOR PROTEIN
Biological speciesCampylobacter jejuni (Campylobacter)
Methodsubtomogram averaging / cryo EM / Resolution: 40.0 Å
AuthorsCohen EJ / Beeby M
Funding support United Kingdom, 1 items
OrganizationGrant numberCountry
Medical Research Council (MRC, United Kingdom) United Kingdom
CitationJournal: Dev Cell / Year: 2024
Title: Evolution of a large periplasmic disk in Campylobacterota flagella enables both efficient motility and autoagglutination.
Authors: Eli J Cohen / Tina Drobnič / Deborah A Ribardo / Aoba Yoshioka / Trishant Umrekar / Xuefei Guo / Jose-Jesus Fernandez / Emma E Brock / Laurence Wilson / Daisuke Nakane / David R Hendrixson / Morgan Beeby /
Abstract: The flagellar motors of Campylobacter jejuni (C. jejuni) and related Campylobacterota (previously epsilonproteobacteria) feature 100-nm-wide periplasmic "basal disks" that have been implicated in ...The flagellar motors of Campylobacter jejuni (C. jejuni) and related Campylobacterota (previously epsilonproteobacteria) feature 100-nm-wide periplasmic "basal disks" that have been implicated in scaffolding a wider ring of additional motor proteins to increase torque, but the size of these disks is excessive for a role solely in scaffolding motor proteins. Here, we show that the basal disk is a flange that braces the flagellar motor during disentanglement of its flagellar filament from interactions with the cell body and other filaments. We show that motor output is unaffected when we shrink or displace the basal disk, and suppressor mutations of debilitated motors occur in flagellar-filament or cell-surface glycosylation pathways, thus sidestepping the need for a flange to overcome the interactions between two flagellar filaments and between flagellar filaments and the cell body. Our results identify unanticipated co-dependencies in the evolution of flagellar motor structure and cell-surface properties in the Campylobacterota.
History
DepositionJun 29, 2023-
Header (metadata) releaseOct 30, 2024-
Map releaseOct 30, 2024-
UpdateJan 1, 2025-
Current statusJan 1, 2025Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_17769.png

Downloads & links

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Map

FileDownload / File: emd_17769.map.gz / Format: CCP4 / Size: 30.5 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationCampylobacter jejuni motor with basal disk genes, flgPQ, expressed at very low level
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
6 Å/pix.
x 200 pix.
= 1200. Å		6 Å/pix.
x 200 pix.
= 1200. Å		6 Å/pix.
x 200 pix.
= 1200. Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 6 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.2
Minimum - Maximum-6.101424 - 4.194216
Average (Standard dev.)0.07755891 (±1.0288466)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions200200200
Spacing200200200
CellA=B=C: 1200.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #1

Fileemd_17769_half_map_1.map
Projections & Slices
AxesZYX

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Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: #2

Fileemd_17769_half_map_2.map
Projections & Slices
AxesZYX

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Histogram (log scale)

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Sample components

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Entire : Campylobacter flagellar motor

EntireName: Campylobacter flagellar motor
Components
  • Organelle or cellular component: Campylobacter flagellar motor

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Supramolecule #1: Campylobacter flagellar motor

SupramoleculeName: Campylobacter flagellar motor / type: organelle_or_cellular_component / ID: 1 / Parent: 0
Details: basal disk genes, flgPQ, expressed at very low level
Source (natural)Organism: Campylobacter jejuni (Campylobacter) / Strain: 81-176 / Organelle: flagellar motor / Location in cell: cell pole

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Experimental details

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Structure determination

Methodcryo EM
Processingsubtomogram averaging
Aggregation statecell

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Sample preparation

BufferpH: 7.2 / Details: Mueller-Hinton broth
GridModel: Quantifoil R2/2 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 60 sec.
VitrificationCryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeTFS GLACIOS
Image recordingFilm or detector model: FEI FALCON IV (4k x 4k) / Average electron dose: 4.0 e/Å2
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.0 µm / Nominal defocus min: 3.0 µm
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN

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Image processing

Final reconstructionAlgorithm: SIMULTANEOUS ITERATIVE (SIRT) / Resolution.type: BY AUTHOR / Resolution: 40.0 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: Dynamo / Number subtomograms used: 95
ExtractionNumber tomograms: 95 / Number images used: 95
Final angle assignmentType: NOT APPLICABLE

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