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- EMDB-10897: Structure of ExbBD complex from pseudomonas savastonoi -

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Basic information

Entry
Database: EMDB / ID: EMD-10897
TitleStructure of ExbBD complex from pseudomonas savastonoi
Map datapostprocesed pseudomonas savastanoi ExbBD complex in LMNG
Sample
  • Complex: ExbB(5)D(2)
    • Protein or peptide: ExbB
    • Protein or peptide: ExbD
Function / homology
Function and homology information


protein import / transmembrane transporter activity / protein transport / plasma membrane
Similarity search - Function
TonB-system energizer ExbB type-1 / TonB system transport protein ExbD type-1 / : / Biopolymer transport protein ExbD/TolR / Biopolymer transport protein ExbD/TolR / MotA/TolQ/ExbB proton channel / MotA/TolQ/ExbB proton channel family
Similarity search - Domain/homology
Biopolymer transport protein ExbB / Biopolymer transport protein ExbD
Similarity search - Component
Biological speciesPseudomonas savastanoi (bacteria)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.8 Å
AuthorsLea SM / Deme JC / Johnson S
Funding support United Kingdom, 4 items
OrganizationGrant numberCountry
Wellcome Trust107929/Z/15/Z United Kingdom
Wellcome Trust100298/Z/12/Z United Kingdom
Wellcome Trust201536/Z/16/Z United Kingdom
Medical Research Council (MRC, United Kingdom)MR/M011984/1 United Kingdom
CitationJournal: Nat Microbiol / Year: 2020
Title: Structures of the stator complex that drives rotation of the bacterial flagellum.
Authors: Justin C Deme / Steven Johnson / Owen Vickery / Amy Aron / Holly Monkhouse / Thomas Griffiths / Rory Hennell James / Ben C Berks / James W Coulton / Phillip J Stansfeld / Susan M Lea /
Abstract: The bacterial flagellum is the prototypical protein nanomachine and comprises a rotating helical propeller attached to a membrane-embedded motor complex. The motor consists of a central rotor ...The bacterial flagellum is the prototypical protein nanomachine and comprises a rotating helical propeller attached to a membrane-embedded motor complex. The motor consists of a central rotor surrounded by stator units that couple ion flow across the cytoplasmic membrane to generate torque. Here, we present the structures of the stator complexes from Clostridium sporogenes, Bacillus subtilis and Vibrio mimicus, allowing interpretation of the extensive body of data on stator mechanism. The structures reveal an unexpected asymmetric AB subunit assembly where the five A subunits enclose the two B subunits. Comparison to structures of other ion-driven motors indicates that this AB architecture is fundamental to bacterial systems that couple energy from ion flow to generate mechanical work at a distance and suggests that such events involve rotation in the motor structures.
History
DepositionApr 22, 2020-
Header (metadata) releaseAug 12, 2020-
Map releaseAug 12, 2020-
UpdateMay 26, 2021-
Current statusMay 26, 2021Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.02
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by cylindrical radius
  • Surface level: 0.02
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_10897.png

Downloads & links

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Map

FileDownload / File: emd_10897.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotationpostprocesed pseudomonas savastanoi ExbBD complex in LMNG
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.82 Å/pix.
x 256 pix.
= 210.432 Å		0.82 Å/pix.
x 256 pix.
= 210.432 Å		0.82 Å/pix.
x 256 pix.
= 210.432 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.822 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.02 / Movie #1: 0.02
Minimum - Maximum-0.045799144 - 0.07278543
Average (Standard dev.)-2.5831278e-06 (±0.003214964)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions256256256
Spacing256256256
CellA=B=C: 210.432 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z0.8220.8220.822
M x/y/z256256256
origin x/y/z0.0000.0000.000
length x/y/z210.432210.432210.432
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS256256256
D min/max/mean-0.0460.073-0.000

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Supplemental data

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Mask #1

Fileemd_10897_msk_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Additional map: relion local resolution filtered pseudomonas savastanoi ExbBD complex...

Fileemd_10897_additional_1.map
Annotationrelion local resolution filtered pseudomonas savastanoi ExbBD complex in LMNG
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Additional map: refinement volume pseudomonas savastanoi ExbBD complex in LMNG

Fileemd_10897_additional_2.map
Annotationrefinement volume pseudomonas savastanoi ExbBD complex in LMNG
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: half map 1

Fileemd_10897_half_map_1.map
Annotationhalf map 1
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: half map 2

Fileemd_10897_half_map_2.map
Annotationhalf map 2
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : ExbB(5)D(2)

EntireName: ExbB(5)D(2)
Components
  • Complex: ExbB(5)D(2)
    • Protein or peptide: ExbB
    • Protein or peptide: ExbD

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Supramolecule #1: ExbB(5)D(2)

SupramoleculeName: ExbB(5)D(2) / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Pseudomonas savastanoi (bacteria)
Recombinant expressionOrganism: Escherichia coli (E. coli)
Molecular weightTheoretical: 195 KDa

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Macromolecule #1: ExbB

MacromoleculeName: ExbB / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO
SequenceString: MTRIQSIASP TRMPRLPRAW RGVAALVLSL MFVPMALADQ SAPTAASAAP AAXATAPVVP DAAAPVQAM EPAAEDNSLG MAHDLSPWGM YQNAEVVVKA VMIGLAIASI ITWTIWISKG F ELLGAKRR LRGEIVNLKK ARSLNEASST ASKEGTLAHL LVHDALEEMR ...String:
MTRIQSIASP TRMPRLPRAW RGVAALVLSL MFVPMALADQ SAPTAASAAP AAXATAPVVP DAAAPVQAM EPAAEDNSLG MAHDLSPWGM YQNAEVVVKA VMIGLAIASI ITWTIWISKG F ELLGAKRR LRGEIVNLKK ARSLNEASST ASKEGTLAHL LVHDALEEMR LSANSREREG IK ERVSFRL ERLVAACGRN MSMGTGVLAT IGSTAPFVGL FGTVWGIMNS FIGIAKTQTT NLA VVAPGI AEALLATALG LVAAIPAVVI YNVFARSIAG YKAQVSDASA HVLLLVSRDL DHLP EPTER NQQQPHMVKV G

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Macromolecule #2: ExbD

MacromoleculeName: ExbD / type: protein_or_peptide / ID: 2 / Enantiomer: LEVO
SequenceString:
MLVKPQRKHG PKAEMNVVPY IDVMLVLLVV FIVTAPLLTN AIPLDLPQTA ATAPLDDAKP VAISIDAKG GLFVDDQPLA EEQLPDALQN LHANDPDVAL TLRADTATDY GRVARVLADV Q RSGITRLA VITESL

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 7.5
Component:
ConcentrationName
100.0 mMTris
150.0 mMNaCl
0.02 %LMNG
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 48.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 0.0003 µm / Nominal defocus min: 0.0001 µm / Nominal magnification: 165000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 1342937
CTF correctionSoftware - Name: SIMPLE (ver. 3.0)
Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.8 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.1) / Number images used: 65617
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: SIMPLE (ver. 3.0)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.1)
Final 3D classificationSoftware - Name: RELION (ver. 3.1)
FSC plot (resolution estimation)

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