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- PDB-8wk0: Cryo-EM structure of a bacterial protein -

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Basic information

Entry
Database: PDB / ID: 8wk0
TitleCryo-EM structure of a bacterial protein
ComponentsHelicase HerA central domain-containing protein
KeywordsIMMUNE SYSTEM / bacterial immune proteins
Function / homologyPHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER
Function and homology information
Biological speciesPaenibacillus sp. 453mf (bacteria)
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.34 Å
AuthorsYu, G. / Liao, F. / Li, X. / Li, Z. / Zhang, H.
Funding support China, 1items
OrganizationGrant numberCountry
National Natural Science Foundation of China (NSFC)32200496 China
CitationJournal: To Be Published
Title: Cryo-EM structure of a bacterial protein
Authors: Yu, G. / Liao, F. / Li, X. / Li, Z. / Zhang, H.
History
DepositionSep 26, 2023Deposition site: PDBJ / Processing site: PDBJ
Revision 1.0Sep 11, 2024Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Helicase HerA central domain-containing protein
D: Helicase HerA central domain-containing protein
M: Helicase HerA central domain-containing protein
N: Helicase HerA central domain-containing protein
O: Helicase HerA central domain-containing protein
L: Helicase HerA central domain-containing protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)468,49616
Polymers465,7586
Non-polymers2,73810
Water00
1


  • Idetical with deposited unit
  • defined by author
  • Evidence: electron microscopy, not applicable
TypeNameSymmetry operationNumber
identity operation1_5551

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Components

#1: Protein
Helicase HerA central domain-containing protein


Mass: 77626.344 Da / Num. of mol.: 6
Source method: isolated from a genetically manipulated source
Details: WP_091160506.1 / Source: (gene. exp.) Paenibacillus sp. 453mf (bacteria) / Gene: SAMN04488601_104156 / Production host: Escherichia coli BL21(DE3) (bacteria)
#2: Chemical
ChemComp-AGS / PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER / ATP-GAMMA-S / ADENOSINE 5'-(3-THIOTRIPHOSPHATE) / ADENOSINE 5'-(GAMMA-THIOTRIPHOSPHATE) / ADENOSINE-5'-DIPHOSPHATE MONOTHIOPHOSPHATE


Mass: 523.247 Da / Num. of mol.: 5 / Source method: obtained synthetically / Formula: C10H16N5O12P3S / Feature type: SUBJECT OF INVESTIGATION / Comment: ATP-gamma-S, energy-carrying molecule analogue*YM
#3: Chemical
ChemComp-MG / MAGNESIUM ION


Mass: 24.305 Da / Num. of mol.: 5 / Source method: obtained synthetically / Formula: Mg / Feature type: SUBJECT OF INVESTIGATION
Has ligand of interestY

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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: Cryo-EM structure of a bacterial protein / Type: COMPLEX / Entity ID: #1 / Source: RECOMBINANT
Molecular weightValue: 0.4 MDa / Experimental value: YES
Source (natural)Organism: Paenibacillus sp. 453mf (bacteria)
Source (recombinant)Organism: Escherichia coli BL21(DE3) (bacteria)
Buffer solutionpH: 7.5
SpecimenEmbedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
VitrificationCryogen name: ETHANE

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
MicroscopyModel: FEI TITAN KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM
Electron lensMode: BRIGHT FIELD / Nominal defocus max: 2500 nm / Nominal defocus min: 500 nm
Image recordingElectron dose: 40 e/Å2 / Detector mode: SUPER-RESOLUTION / Film or detector model: GATAN K3 (6k x 4k)

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Processing

CTF correctionType: PHASE FLIPPING ONLY
3D reconstructionResolution: 3.34 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 117559 / Symmetry type: POINT
Refine LS restraints
Refine-IDTypeDev idealNumber
ELECTRON MICROSCOPYf_bond_d0.00327469
ELECTRON MICROSCOPYf_angle_d0.48737240
ELECTRON MICROSCOPYf_dihedral_angle_d8.6613825
ELECTRON MICROSCOPYf_chiral_restr0.0434284
ELECTRON MICROSCOPYf_plane_restr0.0044731

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