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- EMDB-5633: Cryo-electron tomography of in situ flagellar motor of Borrelia b... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-5633 | |||||||||
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Title | Cryo-electron tomography of in situ flagellar motor of Borrelia burgdorferi FlgE mutant | |||||||||
![]() | Flagellar motor of a FlgE mutant in B. burgdorferi | |||||||||
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![]() | Flagellar motor / assembly / Type III secretion / Lyme disease spirochete / molecular machine | |||||||||
Biological species | ![]() | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 38.0 Å | |||||||||
![]() | Zhao X / Zhang K / Boquoi T / Hu B / Motaleb MA / Miller KA / James ME / Charon NW / Manson MD / Norris SJ ...Zhao X / Zhang K / Boquoi T / Hu B / Motaleb MA / Miller KA / James ME / Charon NW / Manson MD / Norris SJ / Li C / Liu J | |||||||||
![]() | ![]() Title: Cryoelectron tomography reveals the sequential assembly of bacterial flagella in Borrelia burgdorferi. Authors: Xiaowei Zhao / Kai Zhang / Tristan Boquoi / Bo Hu / M A Motaleb / Kelly A Miller / Milinda E James / Nyles W Charon / Michael D Manson / Steven J Norris / Chunhao Li / Jun Liu / ![]() Abstract: Periplasmic flagella are essential for the distinctive morphology, motility, and infectious life cycle of the Lyme disease spirochete Borrelia burgdorferi. In this study, we genetically trapped ...Periplasmic flagella are essential for the distinctive morphology, motility, and infectious life cycle of the Lyme disease spirochete Borrelia burgdorferi. In this study, we genetically trapped intermediates in flagellar assembly and determined the 3D structures of the intermediates to 4-nm resolution by cryoelectron tomography. We provide structural evidence that secretion of rod substrates triggers remodeling of the central channel in the flagellar secretion apparatus from a closed to an open conformation. This open channel then serves as both a gateway and a template for flagellar rod assembly. The individual proteins assemble sequentially to form a modular rod. The hook cap initiates hook assembly on completion of the rod, and the filament cap facilitates filament assembly after formation of the mature hook. Cryoelectron tomography and mutational analysis thus combine synergistically to provide a unique structural blueprint of the assembly process of this intricate molecular machine in intact cells. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 20.8 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 10.9 KB 10.9 KB | Display Display | ![]() |
Images | ![]() | 227.4 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 79.4 KB | Display | ![]() |
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Full document | ![]() | 78.5 KB | Display | |
Data in XML | ![]() | 494 B | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 5627C ![]() 5628C ![]() 5629C ![]() 5630C ![]() 5631C ![]() 5632C C: citing same article ( |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Map
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Annotation | Flagellar motor of a FlgE mutant in B. burgdorferi | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 5.7 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : In situ flagellar motor from B. burgdorferi FlgE mutant cells
Entire | Name: In situ flagellar motor from B. burgdorferi FlgE mutant cells |
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Components |
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-Supramolecule #1000: In situ flagellar motor from B. burgdorferi FlgE mutant cells
Supramolecule | Name: In situ flagellar motor from B. burgdorferi FlgE mutant cells type: sample / ID: 1000 / Number unique components: 20 |
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-Supramolecule #1: flagellar motor
Supramolecule | Name: flagellar motor / type: organelle_or_cellular_component / ID: 1 / Recombinant expression: No / Database: NCBI |
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Source (natural) | Organism: ![]() |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | subtomogram averaging |
Aggregation state | cell |
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Sample preparation
Buffer | Details: BSK-II liquid medium supplemented with 6% rabbit serum |
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Grid | Details: freshly glow-discharged holey carbon grids |
Vitrification | Cryogen name: ETHANE / Instrument: HOMEMADE PLUNGER Method: Bacterial culture was deposited onto freshly glow-discharged, holey carbon grids for 1 min. Grids were blotted with filter paper and then rapidly frozen in liquid ethane, using a homemade ...Method: Bacterial culture was deposited onto freshly glow-discharged, holey carbon grids for 1 min. Grids were blotted with filter paper and then rapidly frozen in liquid ethane, using a homemade gravity-driven plunger apparatus. |
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Electron microscopy
Microscope | FEI POLARA 300 |
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Temperature | Min: 170 K |
Date | Nov 1, 2012 |
Image recording | Number real images: 9831 / Average electron dose: 100 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2 mm / Nominal defocus max: 8.0 µm / Nominal defocus min: 6.0 µm / Nominal magnification: 31000 |
Sample stage | Specimen holder model: OTHER / Tilt series - Axis1 - Min angle: -65 ° / Tilt series - Axis1 - Max angle: 65 ° |
Experimental equipment | ![]() Model: Tecnai Polara / Image courtesy: FEI Company |
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Image processing
Details | The subvolumes of the flagellar motors were extracted computationally from the tomograms and were further aligned in 3D. |
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Final reconstruction | Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 38.0 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: IMOD, Raptor, Protomo / Number subtomograms used: 999 |
Final 3D classification | Number classes: 4 |