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- EMDB-53857: Cryo-EM structure of an extracellular contractile injection syste... -

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Basic information

Entry
Database: EMDB / ID: EMD-53857
TitleCryo-EM structure of an extracellular contractile injection system in Salmonella enterica subspecies salamae, the sheath and inner tube module in extended state.
Map dataSalCIS sheath in extended state helical reconstruction
Sample
  • Complex: Helical complex of the sheath and inner tube module of a contractile injection system
    • Protein or peptide: Phage tail sheath family protein
    • Protein or peptide: Phage tail protein
KeywordsContractile injection system / Salmonella enterica subspecies salamae / CONTRACTILE PROTEIN
Biological speciesSalmonella enterica subsp. salamae (bacteria)
Methodhelical reconstruction / cryo EM / Resolution: 2.28 Å
AuthorsEjaz RN / Tillmann HP / Sofos NH / Siborova M / Taylor NMI
Funding support Denmark, 1 items
OrganizationGrant numberCountry
Independent Research Fund Denmark - Medical Sciences30342400-1201051001 Denmark
CitationJournal: Nat Commun / Year: 2026
Title: Structure of a contractile injection system in Salmonella enterica subsp. salamae.
Authors: Rooshanie N Ejaz / Kristin Funke / Claudia S Kielkopf / Freddie J O Martin / Marta Šiborová / Ivo A Hendriks / Nicholas H Sofos / Tillmann Pape / Eva M Steiner-Rebrova / Michael L Nielsen ...Authors: Rooshanie N Ejaz / Kristin Funke / Claudia S Kielkopf / Freddie J O Martin / Marta Šiborová / Ivo A Hendriks / Nicholas H Sofos / Tillmann Pape / Eva M Steiner-Rebrova / Michael L Nielsen / Marc Erhardt / Nicholas M I Taylor /
Abstract: Extracellular contractile injection systems (eCISs) are phage-derived nanomachines used by bacteria to deliver effectors into target cells. Well-studied examples include the Photorhabdus asymbiotica ...Extracellular contractile injection systems (eCISs) are phage-derived nanomachines used by bacteria to deliver effectors into target cells. Well-studied examples include the Photorhabdus asymbiotica virulence cassettes and the antifeeding prophage from Serratia entomophila, which have been engineered for heterologous cargo delivery. Recent genomic analyses identified eCIS gene clusters in the opportunistic human pathogen Salmonella enterica subspecies salamae, but their structure, function, and biotechnological potential remain unexplored. Here, we report a high-resolution cryo-electron microscopy structure of the S. enterica eCIS. Our atomic models reveal a distinctive sheath architecture, an expansive cage-like shell around a central spike, and an associated integral membrane protein. We identify a putative effector encoded within the operon exhibiting mild periplasmic toxicity and provide evidence that the S. enterica eCIS deviates from canonical eCISs by interacting with the inner membrane. Guided by these structural features, we uncover, to the best of our knowledge, a previously unannotated cluster of contractile injection systems (CISs). Together, our findings expand the known diversity of CISs' structures and functions, and lay the groundwork for engineering customisable protein delivery platforms.
History
DepositionMay 20, 2025-
Header (metadata) releaseApr 22, 2026-
Map releaseApr 22, 2026-
UpdateMay 6, 2026-
Current statusMay 6, 2026Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

Downloads & links

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Map

FileDownload / File: emd_53857.map.gz / Format: CCP4 / Size: 2.7 GB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationSalCIS sheath in extended state helical reconstruction
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.73 Å/pix.
x 900 pix.
= 652.5 Å
0.73 Å/pix.
x 900 pix.
= 652.5 Å
0.73 Å/pix.
x 900 pix.
= 652.5 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.725 Å
Density
Contour LevelBy AUTHOR: 0.0805
Minimum - Maximum-0.29404506 - 0.7332398
Average (Standard dev.)0.0004988392 (±0.023324812)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions900900900
Spacing900900900
CellA=B=C: 652.5 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: SalCIS sheath in extended state half map B

Fileemd_53857_half_map_1.map
AnnotationSalCIS sheath in extended state half map B
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Half map: SalCIS sheath in extended state half map A

Fileemd_53857_half_map_2.map
AnnotationSalCIS sheath in extended state half map A
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Sample components

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Entire : Helical complex of the sheath and inner tube module of a contract...

EntireName: Helical complex of the sheath and inner tube module of a contractile injection system
Components
  • Complex: Helical complex of the sheath and inner tube module of a contractile injection system
    • Protein or peptide: Phage tail sheath family protein
    • Protein or peptide: Phage tail protein

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Supramolecule #1: Helical complex of the sheath and inner tube module of a contract...

SupramoleculeName: Helical complex of the sheath and inner tube module of a contractile injection system
type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Salmonella enterica subsp. salamae (bacteria)

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Macromolecule #1: Phage tail sheath family protein

MacromoleculeName: Phage tail sheath family protein / type: protein_or_peptide / ID: 1 / Number of copies: 18 / Enantiomer: LEVO
Source (natural)Organism: Salmonella enterica subsp. salamae (bacteria)
Molecular weightTheoretical: 57.35193 KDa
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: MGVMKTPGVY IVEQNAFPNS VVEVATAVPA FIGYTEKADN GGKSLSNKGW RITSMSEYRQ YFGGEPQHLF EISEISTTSN ANIREAFKQ SGKTYQITQS NTRHHLYYSM LFFFQNGGGP CYIVSVGNYS DDIDAAVLKG GILPLIKEAE PTMLLIPEAI Q LAEDDCIN ...String:
MGVMKTPGVY IVEQNAFPNS VVEVATAVPA FIGYTEKADN GGKSLSNKGW RITSMSEYRQ YFGGEPQHLF EISEISTTSN ANIREAFKQ SGKTYQITQS NTRHHLYYSM LFFFQNGGGP CYIVSVGNYS DDIDAAVLKG GILPLIKEAE PTMLLIPEAI Q LAEDDCIN VEQAMLGHCG GKMKNRVAIL DVWNGYKDRQ HPDGDCVESF RSKLGTHYLD YAAAYYPWLN TSIVQDSDVS FL NISNIDK LAELLSGEVA LMFSDLEGLS EEELSTGGNK LRATRKQAML DEIAKLSAEI SRPDAVLLHK ILSNMSPLYQ TIM ADIKFQ QNILPPSSAM AGIYTMVDNS RGVWKAPANV SVNAVVSPTV NISDDEQEDL NVTTQGKSIN AIRPFIGEGT LVWG ARTLD GNSVDWRYIN VRRTMIMLEE SIKLASKAYV FEPNVANTWV SMESMLSNFL YGIWKRGGLA GSTPGEAYNV SVGLG KTMT SNDILEGILR ITVLVAMVRP AEFIEITFQQ KMQES

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Macromolecule #2: Phage tail protein

MacromoleculeName: Phage tail protein / type: protein_or_peptide / ID: 2 / Number of copies: 18 / Enantiomer: LEVO
Source (natural)Organism: Salmonella enterica subsp. salamae (bacteria)
Molecular weightTheoretical: 17.091271 KDa
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString:
MADDGSAQSD TVWPMPKFYF EVKWDGGAGA EMVSAFQEVS GLDSEAQPIE YRAGNSPVFS TIKMPGLIKS GNVTLKKGTF KGDNKFYEW YSKIKMNTIA RTAVTINLLD ESGAPVMSWK LKNAWPTKVT GTDLKSDSNE VAVETIELAH EGLEISV

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Experimental details

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Structure determination

Methodcryo EM
Processinghelical reconstruction
Aggregation statehelical array

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Sample preparation

Concentration20 mg/mL
BufferpH: 7.4
GridModel: Quantifoil R2/2 / Mesh: 200 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 2 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 10 sec. / Details: 5mA
VitrificationCryogen name: ETHANE / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeTFS KRIOS
Specialist opticsEnergy filter - Name: TFS Selectris X / Energy filter - Slit width: 10 eV
SoftwareName: EPU
Image recordingFilm or detector model: FEI FALCON IV (4k x 4k) / Average electron dose: 41.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 0.5 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionNumber classes used: 1
Applied symmetry - Helical parameters - Δz: 39.2 Å
Applied symmetry - Helical parameters - Δ&Phi: 21.6 °
Applied symmetry - Helical parameters - Axial symmetry: C6 (6 fold cyclic)
Resolution.type: BY AUTHOR / Resolution: 2.28 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 4) / Number images used: 136877
CTF correctionSoftware - Name: cryoSPARC (ver. 4) / Type: PHASE FLIPPING AND AMPLITUDE CORRECTION
Segment selectionNumber selected: 142404 / Software - Name: cryoSPARC (ver. 4)
Startup modelType of model: NONE
Final angle assignmentType: NOT APPLICABLE

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