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Yorodumi- EMDB-12708: I-Layer C16 at 3.08A - Local refinement with C16 symmetry of I-la... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-12708 | |||||||||||||||
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Title | I-Layer C16 at 3.08A - Local refinement with C16 symmetry of I-layer of the outer membrane core complex from the fully-assembled R388 type IV secretion system. | |||||||||||||||
Map data | I-layer focus refinement C16 symmetry sharpened map | |||||||||||||||
Sample |
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Keywords | type IV secretion system / type 4 secretion system / T4SS / I-layer / core complex / outer membrane complex / R388 plasmid / conjugation / bacterial secretion / secretion / secretion system / protein complex / VirB10 / VirB9 / TrwE / TrwF / MEMBRANE PROTEIN | |||||||||||||||
Function / homology | Function and homology information | |||||||||||||||
Biological species | Escherichia coli (E. coli) | |||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.1 Å | |||||||||||||||
Authors | Mace K / Vadakkepat AK | |||||||||||||||
Funding support | United Kingdom, 4 items
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Citation | Journal: Nature / Year: 2022 Title: Cryo-EM structure of a type IV secretion system. Authors: Kévin Macé / Abhinav K Vadakkepat / Adam Redzej / Natalya Lukoyanova / Clasien Oomen / Nathalie Braun / Marta Ukleja / Fang Lu / Tiago R D Costa / Elena V Orlova / David Baker / Qian Cong ...Authors: Kévin Macé / Abhinav K Vadakkepat / Adam Redzej / Natalya Lukoyanova / Clasien Oomen / Nathalie Braun / Marta Ukleja / Fang Lu / Tiago R D Costa / Elena V Orlova / David Baker / Qian Cong / Gabriel Waksman / Abstract: Bacterial conjugation is the fundamental process of unidirectional transfer of DNAs, often plasmid DNAs, from a donor cell to a recipient cell. It is the primary means by which antibiotic resistance ...Bacterial conjugation is the fundamental process of unidirectional transfer of DNAs, often plasmid DNAs, from a donor cell to a recipient cell. It is the primary means by which antibiotic resistance genes spread among bacterial populations. In Gram-negative bacteria, conjugation is mediated by a large transport apparatus-the conjugative type IV secretion system (T4SS)-produced by the donor cell and embedded in both its outer and inner membranes. The T4SS also elaborates a long extracellular filament-the conjugative pilus-that is essential for DNA transfer. Here we present a high-resolution cryo-electron microscopy (cryo-EM) structure of a 2.8 megadalton T4SS complex composed of 92 polypeptides representing 8 of the 10 essential T4SS components involved in pilus biogenesis. We added the two remaining components to the structural model using co-evolution analysis of protein interfaces, to enable the reconstitution of the entire system including the pilus. This structure describes the exceptionally large protein-protein interaction network required to assemble the many components that constitute a T4SS and provides insights on the unique mechanism by which they elaborate pili. | |||||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_12708.map.gz | 16.2 MB | EMDB map data format | |
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Header (meta data) | emd-12708-v30.xml emd-12708.xml | 18.5 KB 18.5 KB | Display Display | EMDB header |
Images | emd_12708.png | 86.8 KB | ||
Masks | emd_12708_msk_1.map | 17.8 MB | Mask map | |
Filedesc metadata | emd-12708.cif.gz | 6 KB | ||
Others | emd_12708_additional_1.map.gz emd_12708_half_map_1.map.gz emd_12708_half_map_2.map.gz | 15.1 MB 93.7 MB 93.7 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-12708 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-12708 | HTTPS FTP |
-Validation report
Summary document | emd_12708_validation.pdf.gz | 713.9 KB | Display | EMDB validaton report |
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Full document | emd_12708_full_validation.pdf.gz | 713.5 KB | Display | |
Data in XML | emd_12708_validation.xml.gz | 12.2 KB | Display | |
Data in CIF | emd_12708_validation.cif.gz | 13.2 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12708 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12708 | HTTPS FTP |
-Related structure data
Related structure data | 7o3tMC 7o3jC 7o3vC 7o41C 7o42C 7o43C 7oiuC 7q1vC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_12708.map.gz / Format: CCP4 / Size: 17.8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | I-layer focus refinement C16 symmetry sharpened map | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. generated in cubic-lattice coordinate | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.067 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_12708_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Additional map: I-layer focus refinement C16 symmetry unsharpened map
File | emd_12708_additional_1.map | ||||||||||||
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Annotation | I-layer focus refinement C16 symmetry unsharpened map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: I-layer focus refinement C16 symmetry - Half-B
File | emd_12708_half_map_1.map | ||||||||||||
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Annotation | I-layer focus refinement C16 symmetry - Half-B | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: I-layer focus refinement C16 symmetry - Half-A
File | emd_12708_half_map_2.map | ||||||||||||
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Annotation | I-layer focus refinement C16 symmetry - Half-A | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : Type IV secretion system complex
Entire | Name: Type IV secretion system complex |
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Components |
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-Supramolecule #1: Type IV secretion system complex
Supramolecule | Name: Type IV secretion system complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Escherichia coli (E. coli) / Strain: R388 plasmid |
Molecular weight | Theoretical: 2.808 MDa |
-Macromolecule #1: TrwE protein
Macromolecule | Name: TrwE protein / type: protein_or_peptide / ID: 1 / Number of copies: 16 / Enantiomer: LEVO |
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Source (natural) | Organism: Escherichia coli (E. coli) |
Molecular weight | Theoretical: 42.443785 KDa |
Recombinant expression | Organism: Escherichia coli (E. coli) |
Sequence | String: MFGRKKGDVI DAGAELERAE QERIEGEYGA SELASERRPH TPGARTLLMV LLCVIAVVLV TLSYKAYKVR GVVEDDDAQP QQVVRQVIP GYTPRPIRPE PENVPEPPQP TTSVPAIQPA PVTQPVRPQP TGPREKTPYE LARERMLRSG LTAGSGGGED L PRPQGGDV ...String: MFGRKKGDVI DAGAELERAE QERIEGEYGA SELASERRPH TPGARTLLMV LLCVIAVVLV TLSYKAYKVR GVVEDDDAQP QQVVRQVIP GYTPRPIRPE PENVPEPPQP TTSVPAIQPA PVTQPVRPQP TGPREKTPYE LARERMLRSG LTAGSGGGED L PRPQGGDV PAGGLMGGGG GGGELAEKLQ PMRLSGSSAG RLGNRDMLIT QGTQLDCVLE TRLVTTQPGM TTCHLTRDVY ST SGRVVLL DRGSKVVGFY QGGLRQGQAR IFVQWSRIET PSGVVINLDS PGTGPLGEAG LGGWIDRHFW ERFGGAIMIS LIG DLGDWA SRQGSRQGDN SIQFSNTANG VESAAAEALR NSINIPPTLY KNQGERVNIL VARDLDFSDV YSLESIPTK UniProtKB: TrwE protein |
-Macromolecule #2: TrwF protein
Macromolecule | Name: TrwF protein / type: protein_or_peptide / ID: 2 / Number of copies: 16 / Enantiomer: LEVO |
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Source (natural) | Organism: Escherichia coli (E. coli) |
Molecular weight | Theoretical: 29.749586 KDa |
Recombinant expression | Organism: Escherichia coli (E. coli) |
Sequence | String: MKKLAIVALL ASLHAVPALA LDVPSSSRYD HRIRYVTYNP ADVVQVDTVL GVATHIMLEE GEQYLTHAFG DSEAYAFARK GRHIFIKPQ AELANTNLIV VTDRRSYKFR LQMRNDRNGA MYELAFRYPD TQARQTREAN ARAAVEAAFE QRVGAYYNLK Y MMSGDKDI ...String: MKKLAIVALL ASLHAVPALA LDVPSSSRYD HRIRYVTYNP ADVVQVDTVL GVATHIMLEE GEQYLTHAFG DSEAYAFARK GRHIFIKPQ AELANTNLIV VTDRRSYKFR LQMRNDRNGA MYELAFRYPD TQARQTREAN ARAAVEAAFE QRVGAYYNLK Y MMSGDKDI APVNAWDDGR FTYFKFSANA DLPSIYFVDA EGNESLVPRT TVGSSNNIIA VHKVNPKWMI RLGNRALAIF NE AYDPNGV PNDTGTASPA VRRVNKGGN UniProtKB: TrwF protein |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.6 |
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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 57.5 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: OTHER / Imaging mode: OTHER |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Startup model | Type of model: OTHER / Details: CryoSPARC ab-initio |
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Final reconstruction | Applied symmetry - Point group: C16 (16 fold cyclic) / Resolution.type: BY AUTHOR / Resolution: 3.1 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 1280606 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
Final angle assignment | Type: OTHER / Details: Stochastic gradient descent (SGD) |