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- EMDB-30409: CryoEM structure of S.typhimurium flagellar hook-basal body (HBB)... -

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Basic information

Entry
Database: EMDB / ID: EMD-30409
TitleCryoEM structure of S.typhimurium flagellar hook-basal body (HBB) complex
Map dataCryoEM structure of S.typhimurium flagellar hook basal body (HBB) complex
Sample
  • Complex: Hook-basal body complex
Function / homology
Function and homology information


bacterial-type flagellum basal body, distal rod, L ring / bacterial-type flagellum basal body, distal rod, P ring / cytoskeletal motor activity / bacterial-type flagellum-dependent cell motility / cell outer membrane / outer membrane-bounded periplasmic space / structural molecule activity
Similarity search - Function
Flagellar L-ring protein / Flagellar L-ring protein / Flagellar P-ring protein / Flagellar P-ring protein
Similarity search - Domain/homology
Flagellar P-ring protein / Flagellar L-ring protein
Similarity search - Component
Biological speciesSalmonella enterica subsp. enterica serovar Typhimurium (bacteria)
Methodsingle particle reconstruction / cryo EM / Resolution: 6.9 Å
AuthorsYamaguchi T / Makino F / Miyata T / Minamino T / Kato T / Namba K
Funding support Japan, 2 items
OrganizationGrant numberCountry
Japan Society for the Promotion of Science (JSPS)JP25000013, JP18K06155, JP26293097, JP19H03182, JP15H01640 Japan
Japan Agency for Medical Research and Development (AMED)JP19am0101117, JP17pc0101020 Japan
CitationJournal: Nat Commun / Year: 2021
Title: Structure of the molecular bushing of the bacterial flagellar motor.
Authors: Tomoko Yamaguchi / Fumiaki Makino / Tomoko Miyata / Tohru Minamino / Takayuki Kato / Keiichi Namba /
Abstract: The basal body of the bacterial flagellum is a rotary motor that consists of several rings (C, MS and LP) and a rod. The LP ring acts as a bushing supporting the distal rod for its rapid and stable ...The basal body of the bacterial flagellum is a rotary motor that consists of several rings (C, MS and LP) and a rod. The LP ring acts as a bushing supporting the distal rod for its rapid and stable rotation without much friction. Here, we use electron cryomicroscopy to describe the LP ring structure around the rod, at 3.5 Å resolution, from Salmonella Typhimurium. The structure shows 26-fold rotational symmetry and intricate intersubunit interactions of each subunit with up to six partners, which explains the structural stability. The inner surface is charged both positively and negatively. Positive charges on the P ring (the part of the LP ring that is embedded within the peptidoglycan layer) presumably play important roles in its initial assembly around the rod with a negatively charged surface.
History
DepositionJul 28, 2020-
Header (metadata) releaseJun 2, 2021-
Map releaseJun 2, 2021-
UpdateDec 22, 2021-
Current statusDec 22, 2021Processing site: PDBj / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.0185
  • Imaged by UCSF Chimera
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  • Surface view colored by cylindrical radius
  • Surface level: 0.0185
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

Downloads & links

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Map

FileDownload / File: emd_30409.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationCryoEM structure of S.typhimurium flagellar hook basal body (HBB) complex
Voxel sizeX=Y=Z: 1.45 Å
Density
Contour LevelBy AUTHOR: 0.0185 / Movie #1: 0.0185
Minimum - Maximum-0.022580132 - 0.055458326
Average (Standard dev.)0.00079213927 (±0.003525868)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin-199-199-199
Dimensions400400400
Spacing400400400
CellA=B=C: 580.0 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.451.451.45
M x/y/z400400400
origin x/y/z0.0000.0000.000
length x/y/z580.000580.000580.000
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS-199-199-199
NC/NR/NS400400400
D min/max/mean-0.0230.0550.001

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Supplemental data

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Mask #1

Fileemd_30409_msk_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Sample components

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Entire : Hook-basal body complex

EntireName: Hook-basal body complex
Components
  • Complex: Hook-basal body complex

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Supramolecule #1: Hook-basal body complex

SupramoleculeName: Hook-basal body complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#2 / Details: S.typhimurium flagellar motor complex
Source (natural)Organism: Salmonella enterica subsp. enterica serovar Typhimurium (bacteria)
Strain: HK1002

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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: 8
Component:
ConcentrationFormulaName
50.0 mMC4H11NO3Tris-HClTris
50.0 mMNaClSodium chlorideSodium chloride
25.0 mMC3H4N2Imidazole
0.05 % (w/v)C47H88O22Lauryl Maltose Neopentyl Glycol(LMNG)
0.05 % (w/v)Triton X-100
Sugar embeddingMaterial: buffer
GridModel: Quantifoil R1.2/1.3 / Material: MOLYBDENUM / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Pretreatment - Type: GLOW DISCHARGE
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeJEOL CRYO ARM 200
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 100.0 µm / Calibrated defocus max: 8.23 µm / Calibrated defocus min: 0.275 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 1.4 mm / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.5 µm / Nominal magnification: 40000
Specialist opticsEnergy filter - Name: In-column Omega Filter
Sample stageSpecimen holder model: JEOL CRYOSPECPORTER / Cooling holder cryogen: NITROGEN
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Digitization - Frames/image: 1-50 / Number grids imaged: 14370 / Number real images: 12759 / Average exposure time: 10.0 sec. / Average electron dose: 45.0 e/Å2

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Image processing

Particle selectionNumber selected: 64418
Details: particles were picked up by YOLOPick.py (in-house python program)
CTF correctionSoftware - Name: Gctf (ver. 1.06)
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.0 beta-2)
Final 3D classificationNumber classes: 2 / Software - Name: RELION (ver. 3.0 beta-2)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.0 beta-2)
Final reconstructionNumber classes used: 1 / Applied symmetry - Point group: C1 (asymmetric) / Algorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 6.9 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.0 beta-2) / Number images used: 14370
FSC plot (resolution estimation)

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Atomic model buiding 1

RefinementSpace: REAL / Protocol: OTHER / Overall B value: 38

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