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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-13154 | |||||||||||||||
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Title | 2.29 A Mycobacterium tuberculosis EspB. | |||||||||||||||
![]() | Local B-factor sharpened map by Locspiral. | |||||||||||||||
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![]() | Cryo-EM / EspB / ESX-1 / Preferential orientation / PROTEIN TRANSPORT | |||||||||||||||
Function / homology | ESX-1 secretion-associated protein EspB, PE domain / ESX-1 secreted protein B PE domain / protein secretion by the type VII secretion system / PPE superfamily / biological process involved in interaction with host / extracellular region / identical protein binding / ESX-1 secretion-associated protein EspB![]() | |||||||||||||||
Biological species | ![]() | |||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.29 Å | |||||||||||||||
![]() | Gijsbers A / Zhang Y | |||||||||||||||
Funding support | ![]() ![]()
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![]() | ![]() Title: Priming mycobacterial ESX-secreted protein B to form a channel-like structure. Authors: Abril Gijsbers / Vanesa Vinciauskaite / Axel Siroy / Ye Gao / Giancarlo Tria / Anjusha Mathew / Nuria Sánchez-Puig / Carmen López-Iglesias / Peter J Peters / Raimond B G Ravelli / ![]() ![]() Abstract: ESX-1 is a major virulence factor of , a secretion machinery directly involved in the survival of the microorganism from the immune system defence. It disrupts the phagosome membrane of the host cell ...ESX-1 is a major virulence factor of , a secretion machinery directly involved in the survival of the microorganism from the immune system defence. It disrupts the phagosome membrane of the host cell through a contact-dependent mechanism. Recently, the structure of the inner-membrane core complex of the homologous ESX-3 and ESX-5 was resolved; however, the elements involved in the secretion through the outer membrane or those acting on the host cell membrane are unknown. Protein substrates might form this missing element. Here, we describe the oligomerisation process of the ESX-1 substrate EspB, which occurs upon cleavage of its C-terminal region and is favoured by an acidic environment. Cryo-electron microscopy data shows that quaternary structure of EspB is conserved across slow growing species, but not in the fast growing . EspB assembles into a channel with dimensions and characteristics suitable for the transit of ESX-1 substrates, as shown by the presence of another EspB trapped within. Our results provide insight into the structure and assembly of EspB, and suggests a possible function as a structural element of ESX-1. | |||||||||||||||
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 | ![]() | 11.3 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 21.5 KB 21.5 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 9.1 KB | Display | ![]() |
Images | ![]() | 212.5 KB | ||
Masks | ![]() | 64 MB | ![]() | |
Filedesc metadata | ![]() | 6.4 KB | ||
Others | ![]() ![]() ![]() | 7 MB 49.6 MB 49.6 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7p13MC ![]() 7p0zC M: atomic model generated by this map 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 | Local B-factor sharpened map by Locspiral. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.834 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
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Projections & Slices |
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Density Histograms |
-Additional map: Global B-factor sharpened map from Relion postprocessing.
File | emd_13154_additional_1.map | ||||||||||||
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Annotation | Global B-factor sharpened map from Relion postprocessing. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Unmasked halfmap 1.
File | emd_13154_half_map_1.map | ||||||||||||
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Annotation | Unmasked halfmap 1. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Unmasked halfmap 2.
File | emd_13154_half_map_2.map | ||||||||||||
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Annotation | Unmasked halfmap 2. | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : heptamer of EspB
Entire | Name: heptamer of EspB |
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Components |
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-Supramolecule #1: heptamer of EspB
Supramolecule | Name: heptamer of EspB / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 220 KDa |
-Macromolecule #1: ESX-1 secretion-associated protein EspB
Macromolecule | Name: ESX-1 secretion-associated protein EspB / type: protein_or_peptide / ID: 1 / Number of copies: 7 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 31.561842 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: SMHTQSQTVT VDQQEILNRA NEVEAPMADP PTDVPITPCE LTAAKNAAQQ LVLSADNMRE YLAAGAKERQ RLATSLRNAA KAYGEVDEE AATALDNDGE GTVQAESAGA VGGDSSAELT DTPRVATAGE PNFMDLKEAA RKLETGDQGA SLAHFADGWN T FNLTLQGD ...String: SMHTQSQTVT VDQQEILNRA NEVEAPMADP PTDVPITPCE LTAAKNAAQQ LVLSADNMRE YLAAGAKERQ RLATSLRNAA KAYGEVDEE AATALDNDGE GTVQAESAGA VGGDSSAELT DTPRVATAGE PNFMDLKEAA RKLETGDQGA SLAHFADGWN T FNLTLQGD VKRFRGFDNW EGDAATACEA SLDQQRQWIL HMAKLSAAMA KQAQYVAQLH VWARREHPTY EDIVGLERLY AE NPSARDQ ILPVYAEYQQ RSEKVLTEYN NKAALEPVNP PKPPPAIKID PP UniProtKB: ESX-1 secretion-associated protein EspB |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 6 mg/mL | |||||||||
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Buffer | pH: 5.5 Component:
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Grid | Model: UltrAuFoil R1.2/1.3 / Material: GOLD / Mesh: 300 / Support film - Material: GOLD / Support film - topology: HOLEY ARRAY / Support film - Film thickness: 2000 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 40 sec. / Pretreatment - Atmosphere: AIR / Pretreatment - Pressure: 0.01 kPa | |||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV | |||||||||
Details | This sample was monodisperse. |
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Electron microscopy
Microscope | TFS KRIOS |
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Temperature | Min: 90.0 K |
Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
Details | Basic direct alignments were done as well as astigmatism and coma alignment using AutoCTF |
Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Digitization - Dimensions - Width: 5760 pixel / Digitization - Dimensions - Height: 4092 pixel / Number grids imaged: 1 / Number real images: 2334 / Average exposure time: 1.8 sec. / Average electron dose: 40.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | C2 aperture diameter: 50.0 µm / Calibrated magnification: 105000 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal magnification: 105000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |