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- PDB-9jsg: Clostridium perfringens iota toxin pore Ib in pore state -

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Basic information

Entry
Database: PDB / ID: 9jsg
TitleClostridium perfringens iota toxin pore Ib in pore state
ComponentsIota toxin component Ib
KeywordsTOXIN / Prepore / Pre-pore / Pore / Translocation
Function / homology
Function and homology information


protein homooligomerization / extracellular region
Similarity search - Function
Bacterial exotoxin B / Protective antigen, heptamerisation domain / Protective antigen, Ca-binding domain / Clostridial binary toxin B/anthrax toxin PA, domain 3 / Protective antigen, heptamerisation domain superfamily / Clostridial binary toxin B/anthrax toxin PA Ca-binding domain / Clostridial binary toxin B/anthrax toxin PA domain 2 / Clostridial binary toxin B/anthrax toxin PA domain 3 / PA14 / PA14 domain
Similarity search - Domain/homology
Iota toxin component Ib
Similarity search - Component
Biological speciesClostridium perfringens (bacteria)
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.47 Å
AuthorsYamada, T. / Sugita, Y. / Yoshida, T. / Noda, T. / Tsuge, H.
Funding support Japan, 4items
OrganizationGrant numberCountry
Japan Society for the Promotion of Science (JSPS)21J13410 Japan
Japan Society for the Promotion of Science (JSPS)21H02452 Japan
Japan Society for the Promotion of Science (JSPS)24K01993 Japan
Japan Agency for Medical Research and Development (AMED)2366 Japan
CitationJournal: To Be Published
Title: Step-by-Step Maturation Mechanism of Binary Toxin Pore Revealed by Cryo-EM Analysis
Authors: Yamada, T. / Sugita, Y. / Yoshida, T. / Noda, T. / Tsuge, H.
History
DepositionSep 30, 2024Deposition site: PDBJ / Processing site: PDBJ
Revision 1.0May 14, 2025Provider: repository / Type: Initial release
Revision 1.0May 14, 2025Data content type: EM metadata / Data content type: EM metadata / Provider: repository / Type: Initial release
Revision 1.0May 14, 2025Data content type: Additional map / Part number: 1 / Data content type: Additional map / Provider: repository / Type: Initial release
Revision 1.0May 14, 2025Data content type: Half map / Part number: 1 / Data content type: Half map / Provider: repository / Type: Initial release
Revision 1.0May 14, 2025Data content type: Half map / Part number: 2 / Data content type: Half map / Provider: repository / Type: Initial release
Revision 1.0May 14, 2025Data content type: Image / Data content type: Image / Provider: repository / Type: Initial release
Revision 1.0May 14, 2025Data content type: Primary map / Data content type: Primary map / Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: Iota toxin component Ib
hetero molecules


Theoretical massNumber of molelcules
Total (without water)76,7594
Polymers76,6381
Non-polymers1203
Water00
1
A: Iota toxin component Ib
hetero molecules

A: Iota toxin component Ib
hetero molecules

A: Iota toxin component Ib
hetero molecules

A: Iota toxin component Ib
hetero molecules

A: Iota toxin component Ib
hetero molecules

A: Iota toxin component Ib
hetero molecules

A: Iota toxin component Ib
hetero molecules


Theoretical massNumber of molelcules
Total (without water)537,31028
Polymers536,4687
Non-polymers84221
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
point symmetry operation6
Noncrystallographic symmetry (NCS)NCS oper:
IDCodeMatrixVector
1given(1), (1), (1)
2generate(0.62349, -0.781832), (0.781832, 0.62349), (1)135.57231, -47.4388
3generate(-0.222521, -0.974928), (0.974928, -0.222521), (1)257.18939, 28.97829
4generate(-0.900969, -0.433884), (0.433884, -0.900969), (1)273.27115, 171.70764
5generate(-0.900969, 0.433884), (-0.433884, -0.900969), (1)171.70764, 273.27115
6generate(-0.222521, 0.974928), (-0.974928, -0.222521), (1)28.97829, 257.18939
7generate(0.62349, 0.781832), (-0.781832, 0.62349), (1)-47.4388, 135.57231

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Components

#1: Protein Iota toxin component Ib


Mass: 76638.297 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Clostridium perfringens (bacteria) / Strain: NCIB 10748 / Plasmid: pET23a / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: Q46221
#2: Chemical ChemComp-CA / CALCIUM ION


Mass: 40.078 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: Ca / Feature type: SUBJECT OF INVESTIGATION
Has ligand of interestY
Has protein modificationN

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Experimental details

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Experiment

ExperimentMethod: ELECTRON MICROSCOPY
EM experimentAggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction

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Sample preparation

ComponentName: Clostridium perfringens iota toxin component Ib / Type: ORGANELLE OR CELLULAR COMPONENT / Entity ID: #1 / Source: RECOMBINANT
Molecular weightExperimental value: NO
Source (natural)Organism: Clostridium perfringens (bacteria) / Strain: NCIB 10748
Source (recombinant)Organism: Escherichia coli BL21(DE3) (bacteria) / Plasmid: pet23a
Buffer solutionpH: 8
Buffer component
IDConc.NameFormulaBuffer-ID
120 mMtris hydroxymethyl aminomethaneC4H11NO31
2100 mMSodium ChlorideNaCl1
31.0 mMCalcium ChlorideCaCl21
SpecimenEmbedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
Specimen supportGrid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R1.2/1.3
VitrificationInstrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277.15 K

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
MicroscopyModel: TFS KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM
Electron lensMode: BRIGHT FIELD / Nominal defocus max: 1600 nm / Nominal defocus min: 600 nm / Cs: 0.01 mm
Specimen holderCryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER
Image recordingElectron dose: 50 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) / Num. of grids imaged: 1 / Num. of real images: 11549
EM imaging opticsEnergyfilter name: GIF Bioquantum / Energyfilter slit width: 20 eV

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Processing

EM software
IDNameVersionCategoryDetails
1cryoSPARC3.3.2particle selectiontemplate picker
4cryoSPARC3.3.2CTF correctionpatch CTF estimation
7UCSF ChimeraX1.2.4model fitting
9PHENIX1.19.2model refinement
10Coot0.9.5model refinement
12cryoSPARC3.3.2final Euler assignment
13cryoSPARC3.3.2classification
14cryoSPARC3.3.23D reconstruction
CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
Particle selectionNum. of particles selected: 6223563
SymmetryPoint symmetry: C7 (7 fold cyclic)
3D reconstructionResolution: 2.47 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 161249 / Symmetry type: POINT
Atomic model buildingSpace: REAL
Atomic model buildingPDB-ID: 6klx

6klx


Pdb chain-ID: A / Accession code: 6klx / Source name: PDB / Type: experimental model

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