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- PDB-9lj4: CryoEM Structures Uncover the Unexpected Hinges of IscB for Enhan... -

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Basic information

Entry
Database: PDB / ID: 9lj4
TitleCryoEM Structures Uncover the Unexpected Hinges of IscB for Enhanced Gene Editing
Components
  • DNA (5'-D(P*TP*AP*GP*GP*TP*GP*GP*TP*TP*GP*TP*CP*GP*GP*CP*CP*A)-3')
  • DNA (5'-D(P*TP*CP*AP*AP*CP*CP*AP*CP*CP*TP*AP*C)-3')
  • Iscb+twin-strep-tag
  • RNA (190-MER)
KeywordsDNA BINDING PROTEIN / IscB / Target DNA / HNH
Function / homologyDNA / DNA (> 10) / RNA / RNA (> 10) / RNA (> 100)
Function and homology information
Biological speciessynthetic construct (others)
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.09 Å
AuthorsHu, C.Y. / Wang, F.Z. / Ma, S.S. / Zhang, S.F.
Funding support Singapore, 1items
OrganizationGrant numberCountry
Ministry of Education (MoE, Singapore) Singapore
CitationJournal: To Be Published
Title: CryoEM Structures Uncover the Unexpected Hinges of IscB for Enhanced Gene Editing
Authors: Hu, C.Y. / Wang, F.Z. / Ma, S.S. / Zhang, S.F.
History
DepositionJan 14, 2025Deposition site: PDBJ / Processing site: PDBJ
Revision 1.0Jan 21, 2026Provider: repository / Type: Initial release
Revision 1.0Jan 21, 2026Data content type: EM metadata / Data content type: EM metadata / Provider: repository / Type: Initial release
Revision 1.0Jan 21, 2026Data content type: Half map / Part number: 1 / Data content type: Half map / Provider: repository / Type: Initial release
Revision 1.0Jan 21, 2026Data content type: Half map / Part number: 2 / Data content type: Half map / Provider: repository / Type: Initial release
Revision 1.0Jan 21, 2026Data content type: Image / Data content type: Image / Provider: repository / Type: Initial release
Revision 1.0Jan 21, 2026Data content type: Primary map / Data content type: Primary map / Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Iscb+twin-strep-tag
B: RNA (190-MER)
C: DNA (5'-D(P*TP*CP*AP*AP*CP*CP*AP*CP*CP*TP*AP*C)-3')
D: DNA (5'-D(P*TP*AP*GP*GP*TP*GP*GP*TP*TP*GP*TP*CP*GP*GP*CP*CP*A)-3')
hetero molecules


Theoretical massNumber of molelcules
Total (without water)132,4297
Polymers132,3154
Non-polymers1143
Water00
1


  • Idetical with deposited unit
  • defined by author&software
  • Evidence: electron microscopy, not applicable
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1

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Components

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DNA chain , 2 types, 2 molecules CD

#3: DNA chain DNA (5'-D(P*TP*CP*AP*AP*CP*CP*AP*CP*CP*TP*AP*C)-3')


Mass: 3551.346 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) synthetic construct (others) / Production host: synthetic construct (others)
#4: DNA chain DNA (5'-D(P*TP*AP*GP*GP*TP*GP*GP*TP*TP*GP*TP*CP*GP*GP*CP*CP*A)-3')


Mass: 5274.406 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) synthetic construct (others) / Production host: synthetic construct (others)

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Protein / RNA chain , 2 types, 2 molecules AB

#1: Protein Iscb+twin-strep-tag


Mass: 62165.832 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) synthetic construct (others) / Production host: Escherichia coli (E. coli)
#2: RNA chain RNA (190-MER)


Mass: 61323.492 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) synthetic construct (others) / Production host: Escherichia coli (E. coli)

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Non-polymers , 2 types, 3 molecules

#5: Chemical ChemComp-ZN / ZINC ION


Mass: 65.409 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: Zn / Feature type: SUBJECT OF INVESTIGATION
#6: Chemical ChemComp-MG / MAGNESIUM ION


Mass: 24.305 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: Mg / Feature type: SUBJECT OF INVESTIGATION

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Details

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: The ternary complex formed by IscB, wRNA, and Target DNA
Type: COMPLEX / Entity ID: #2-#4 / Source: RECOMBINANT
Source (natural)Organism: synthetic construct (others)
Source (recombinant)Organism: Escherichia coli (E. coli)
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: TFS KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM
Electron lensMode: DIFFRACTION / Nominal defocus max: 2500 nm / Nominal defocus min: 1000 nm
Image recordingElectron dose: 49.34 e/Å2 / Film or detector model: GATAN K3 (6k x 4k)

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Processing

EM software
IDNameVersionCategory
1cryoSPARCparticle selection
2PHENIX1.20.1_4487model refinement
13cryoSPARC3D reconstruction
CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
3D reconstructionResolution: 3.09 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 75033 / Symmetry type: POINT
RefinementHighest resolution: 3.09 Å
Stereochemistry target values: REAL-SPACE (WEIGHTED MAP SUM AT ATOM CENTERS)
Refine LS restraints
Refine-IDTypeDev idealNumber
ELECTRON MICROSCOPYf_bond_d0.0059271
ELECTRON MICROSCOPYf_angle_d0.82613559
ELECTRON MICROSCOPYf_dihedral_angle_d24.2853655
ELECTRON MICROSCOPYf_chiral_restr0.0421645
ELECTRON MICROSCOPYf_plane_restr0.006927

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