[English] 日本語
Yorodumi- EMDB-12709: Stalk C1 at 3.71A - Local refinement without symmetry of the Stal... -
+
Open data
-
Basic information
| Entry | ![]() | |||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Title | Stalk C1 at 3.71A - Local refinement without symmetry of the Stalk complex from the fully-assembled R388 type IV secretion system. | |||||||||||||||
Map data | Stalk local refinement C1 sharpened map | |||||||||||||||
Sample |
| |||||||||||||||
Keywords | type IV secretion system / type 4 secretion system / T4SS / stalk / stalk complex / inner membrane / R388 plasmid / conjugation / bacterial secretion / secretion / secretion system / protein complex / VirB5 / VirB6 / TrwJ / TrwI / MEMBRANE PROTEIN | |||||||||||||||
| Function / homology | Type IV secretion system, VirB5 / Type IV secretion system, VirB5-domain / Type IV secretion system proteins / Plasmid conjugal transfer TrbL/VirB6 / TrbL/VirB6 plasmid conjugal transfer protein / protein secretion by the type IV secretion system / membrane / TrwJ protein / TrwI protein Function and homology information | |||||||||||||||
| Biological species | ![]() | |||||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.7 Å | |||||||||||||||
Authors | Mace K / Vadakkepat AK | |||||||||||||||
| Funding support | United Kingdom, 4 items
| |||||||||||||||
Citation | Journal: Nature / Year: 2022Title: 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 |
|
-
Structure visualization
| Supplemental images |
|---|
-
Downloads & links
-EMDB archive
| Map data | emd_12709.map.gz | 10.1 MB | EMDB map data format | |
|---|---|---|---|---|
| Header (meta data) | emd-12709-v30.xml emd-12709.xml | 18.3 KB 18.3 KB | Display Display | EMDB header |
| Images | emd_12709.png | 48.9 KB | ||
| Masks | emd_12709_msk_1.map | 10.9 MB | Mask map | |
| Filedesc metadata | emd-12709.cif.gz | 5.7 KB | ||
| Others | emd_12709_additional_1.map.gz emd_12709_half_map_1.map.gz emd_12709_half_map_2.map.gz | 9.9 MB 10.1 MB 10.1 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-12709 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-12709 | HTTPS FTP |
-Validation report
| Summary document | emd_12709_validation.pdf.gz | 749.5 KB | Display | EMDB validaton report |
|---|---|---|---|---|
| Full document | emd_12709_full_validation.pdf.gz | 749.1 KB | Display | |
| Data in XML | emd_12709_validation.xml.gz | 7 KB | Display | |
| Data in CIF | emd_12709_validation.cif.gz | 8.8 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12709 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12709 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 7o3vMC ![]() 7o3jC ![]() 7o3tC ![]() 7o41C ![]() 7o42C ![]() 7o43C ![]() 7oiuC ![]() 7q1vC M: atomic model generated by this map C: citing same article ( |
|---|---|
| Similar structure data | Similarity search - Function & homology F&H Search |
-
Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
|---|
-
Map
| File | Download / File: emd_12709.map.gz / Format: CCP4 / Size: 10.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Annotation | Stalk local refinement C1 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 |
| ||||||||||||||||||||||||||||||||||||
| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
|
-Supplemental data
-Mask #1
| File | emd_12709_msk_1.map | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Projections & Slices |
| ||||||||||||
| Density Histograms |
-Additional map: Stalk local refinement C1 unsharpened map
| File | emd_12709_additional_1.map | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Annotation | Stalk local refinement C1 unsharpened map | ||||||||||||
| Projections & Slices |
| ||||||||||||
| Density Histograms |
-Half map: Stalk local refinement C1 - Half-B
| File | emd_12709_half_map_1.map | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Annotation | Stalk local refinement C1 - Half-B | ||||||||||||
| Projections & Slices |
| ||||||||||||
| Density Histograms |
-Half map: Stalk local refinement C1 - Half-A
| File | emd_12709_half_map_2.map | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Annotation | Stalk local refinement C1 - Half-A | ||||||||||||
| Projections & Slices |
| ||||||||||||
| Density Histograms |
-
Sample components
-Entire : Type IV secretion system complex
| Entire | Name: Type IV secretion system complex |
|---|---|
| Components |
|
-Supramolecule #1: Type IV secretion system complex
| Supramolecule | Name: Type IV secretion system complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
|---|---|
| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 2.808 MDa |
-Macromolecule #1: TrwJ protein
| Macromolecule | Name: TrwJ protein / type: protein_or_peptide / ID: 1 / Number of copies: 5 / Enantiomer: LEVO |
|---|---|
| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 25.190461 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: MKKLVMTAAV AAILGAASPV MAQGIPVFDG TRALDFVQQF ARMKEQLDTA KDQLAEAQRM YEAVTGGRGL GDLMRNAQLR EYLPDDLRT VYDSANGGGY SGISGSINDI LRDERLNGSV ADMRRSIEER SRTAAATDKA VGLRAYEGAQ QRLAQIEGLM D EISRTQDQ ...String: MKKLVMTAAV AAILGAASPV MAQGIPVFDG TRALDFVQQF ARMKEQLDTA KDQLAEAQRM YEAVTGGRGL GDLMRNAQLR EYLPDDLRT VYDSANGGGY SGISGSINDI LRDERLNGSV ADMRRSIEER SRTAAATDKA VGLRAYEGAQ QRLAQIEGLM D EISRTQDQ KAIEELQARI AGEQAAIQNE TTKLQMIAQL RQAEQALISE QRRERNMRIL SSGNQGMPTI Q UniProtKB: TrwJ protein |
-Macromolecule #2: TrwI protein
| Macromolecule | Name: TrwI protein / type: protein_or_peptide / ID: 2 / Number of copies: 5 / Enantiomer: LEVO |
|---|---|
| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 35.324172 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: MAFELFTPLF NKIDQTTATY VTDISSRAIA AITPVVSVGL TLGFITYGWL IIRGAVEMPV AEFLNRCLRI GIIVSIALAG GLYQGEIAN AITTVPDELA SALLGNPTQG ASAAALVDQS AQQGFDRASE AFEEAGFFSS DGLLYGLFGI IILLATGLLA A IGGAFLLL ...String: MAFELFTPLF NKIDQTTATY VTDISSRAIA AITPVVSVGL TLGFITYGWL IIRGAVEMPV AEFLNRCLRI GIIVSIALAG GLYQGEIAN AITTVPDELA SALLGNPTQG ASAAALVDQS AQQGFDRASE AFEEAGFFSS DGLLYGLFGI IILLATGLLA A IGGAFLLL AKIALALLAG LGPLFILALI WQPTHRFFDQ WAQQVLNYGL LIVLFAAVFG LLMQIFGSYM ADLRFDGAQN VA YAIGGSV ILSIVSIVLL MQLPSIASGL AGGIGLGYMW ELRSMRSGAG AAMRGGRAMA RGARAAPGAA RGAAVGAANM AKT VATGGA GVARAAAGYF RGRKAG UniProtKB: TrwI protein |
-Experimental details
-Structure determination
| Method | cryo EM |
|---|---|
Processing | single particle reconstruction |
| Aggregation state | particle |
-
Sample preparation
| Buffer | pH: 7.6 |
|---|---|
| Vitrification | Cryogen name: ETHANE |
-
Electron microscopy
| Microscope | FEI TITAN KRIOS |
|---|---|
| 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 |
|---|---|
| Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.7 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 566815 |
| Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
| Final angle assignment | Type: OTHER / Details: Stochastic gradient descent (SGD) |
Movie
Controller
About Yorodumi



Keywords
Authors
United Kingdom, 4 items
Citation



















Z (Sec.)
Y (Row.)
X (Col.)




















































FIELD EMISSION GUN
