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- EMDB-16792: Photorhabdus luminescens TcdA1 prepore-to-pore intermediate, K567... -

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Basic information

Entry
Database: EMDB / ID: EMD-16792
TitlePhotorhabdus luminescens TcdA1 prepore-to-pore intermediate, K567W K2008W mutant
Map dataThe local filtered map that was used for model building
Sample
  • Complex: TcdA1
    • Protein or peptide: TcdA1
KeywordsBacterial toxin / Tc toxin / TOXIN
Function / homology
Function and homology information


identical protein binding
Similarity search - Function
ABC toxin, N-terminal domain / ABC toxin N-terminal region / TcA receptor binding domain / TcA receptor binding domain / Insecticidal toxin complex/plasmid virulence protein / Tc toxin complex TcA, C-terminal TcB-binding domain / Neuraminidase-like domain / Salmonella virulence plasmid 28.1kDa A protein / Tc toxin complex TcA C-terminal TcB-binding domain / Neuraminidase-like domain
Similarity search - Domain/homology
Biological speciesPhotorhabdus luminescens (bacteria)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.2 Å
AuthorsNganga PN / Roderer D / Belyy A / Prumbaum D / Raunser S
Funding support Germany, 1 items
OrganizationGrant numberCountry
Max Planck Society Germany
CitationJournal: to be published
Title: Kinetics of the syringe-like injection mechanism of Tc toxins
Authors: Nganga PN / Folz J / Kucher S / Roderer D / Xu Y / Sitsel O / Belyy A / Prumbaum D / Assafa TE / Kuehnemuth R / Dong M / Seidel C / Bordignon E / Raunser S
History
DepositionMar 3, 2023-
Header (metadata) releaseMar 13, 2024-
Map releaseMar 13, 2024-
UpdateMar 13, 2024-
Current statusMar 13, 2024Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

Downloads & links

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Map

FileDownload / File: emd_16792.map.gz / Format: CCP4 / Size: 669.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationThe local filtered map that was used for model building
Voxel sizeX=Y=Z: 0.88 Å
Density
Contour LevelBy AUTHOR: 0.015
Minimum - Maximum-0.051121328 - 0.12087269
Average (Standard dev.)0.00024732033 (±0.0029014912)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions560560560
Spacing560560560
CellA=B=C: 492.8 Å
α=β=γ: 90.0 °

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Supplemental data

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Mask #1

Fileemd_16792_msk_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Half map: First half map

Fileemd_16792_half_map_1.map
AnnotationFirst half map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Half map: Second half map

Fileemd_16792_half_map_2.map
AnnotationSecond half map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Sample components

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Entire : TcdA1

EntireName: TcdA1
Components
  • Complex: TcdA1
    • Protein or peptide: TcdA1

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Supramolecule #1: TcdA1

SupramoleculeName: TcdA1 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Photorhabdus luminescens (bacteria)
Molecular weightTheoretical: 1.5 MDa

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Macromolecule #1: TcdA1

MacromoleculeName: TcdA1 / type: protein_or_peptide / ID: 1 / Number of copies: 5 / Enantiomer: LEVO
Source (natural)Organism: Photorhabdus luminescens (bacteria)
Molecular weightTheoretical: 285.489844 KDa
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: MAHHHHHHSS GLEVLFQGPM NESVKEIPDV LKSQCGFNCL TDISHSSFNE FRQQVSEHLS WSETHDLYHD AQQAQKDNRL YEARILKRA NPQLQNAVHL AILAPNAELI GYNNQFSGRA SQYVAPGTVS SMFSPAAYLT ELYREARNLH ASDSVYYLDT R RPDLKSMA ...String:
MAHHHHHHSS GLEVLFQGPM NESVKEIPDV LKSQCGFNCL TDISHSSFNE FRQQVSEHLS WSETHDLYHD AQQAQKDNRL YEARILKRA NPQLQNAVHL AILAPNAELI GYNNQFSGRA SQYVAPGTVS SMFSPAAYLT ELYREARNLH ASDSVYYLDT R RPDLKSMA LSQQNMDIEL STLSLSNELL LESIKTESKL ENYTKVMEML STFRPSGATP YHDAYENVRE VIQLQDPGLE QL NASPAIA GLMHQASLLG INASISPELF NILTEEITEG NAEELYKKNF GNIEPASLAM PEYLKRYYNL SDEELSQFIG KAS NFGQQE YSNNQLITPV VNSSDGTVKV YRITREYTTN AYQMDVELFP FGGENYRLDY KFKNFYNASY LSIKLNDKRE LVRT EGAPQ VNIEYSANIT LNTADISQPF EIGLTRVLPS GSWAYAAAKF TVEEYNQYSF LLKLNKAIRL SRATELSPTI LEGIV RSVN LQLDINTDVL GKVFLTKYYM QRYAIHAETA LILCNAPISQ RSYDNQPSQF DRLFNTPLLN GQYFSTGDEE IDLNSG STG DWRKTILKRA FNIDDVSLFR LLWITDHDNK DGKIKNNLKN LSNLYIGKLL ADIHQLTIDE LDLLLIAVGE GKTNLSA IS DKQLATLIRK LNTITSWLHT QKWSVFQLFI MTSTSYNKTL TPEIKNLLDT VYHGLQGFDK DKADLLHVMA PYIAATLQ L SSENVAHSVL LWADKLQPGD GAMTAEKFWD WLNTKYTPGS SEAVETQEHI VQYCQALAQL EMVYHSTGIN ENAFRLFVT KPEMFGAATG AAPAHDALSL IMLTRFADWV NALGEKASSV LAAFEANSLT AEQLADAMNL DANLLLQASI QAQNHQHLPP VTPENAFSC WTSINTILQW VNVAQQLNVA PQGVSALVGL DYIQSMKETP TYAQWENAAG VLTAGLNSQQ ANTLHAFLDE S RSAALSTY YIRQVAKAAA AIKSRDDLYQ YLLIDNQVSA AIKTTRIAEA IASIQLYVNR ALENVEENAN SGVISRQFFI DW DKYNKRY STWAGVSQLV YYPENYIDPT MRIGQTKMMD ALLQSVSQSQ LNADTVEDAF MSYLTSFEQV ANLKVISAYH DNI NNDQGL TYFIGLSETD AGEYYWRSVD HSKFNDGKFA ANAWSEWHKI DCPINPYKST IRPVIYKSRL YLLWLEQKEI TKQT GNSKD GYQTETDYRY ELKLAHIRYD GTWNTPITFD VNKKISELKL EKNRAPGLYC AGYQGEDTLL VMFYNQQDTL DSYKN ASMQ GLYIFADMAS KDMTPEQSNV YRDNSYQQFD TNNVRRVNNR YAEDYEIPSS VSSRKDYGWG DYYLSMVYNG DIPTIN YKA ASSDLKIYIS PKLRIIHNGY EGQKRNQCNL MNKYGKLGDK FIVYTSLGVN PNNSSNKLMF YPVYQYSGNT SGLNQGR LL FHRDTTYPSK VEAWIPGAKR SLTNQNAAIG DDYATDSLNK PDDLKQYIFM TDSKGTATDV SGPVEINTAI SPAKVQII V KAGGKEQTFT ADKDVSIQPS PSFDEMNYQF NALEIDGSGL NFINNSASID VTFTAFAEDG RKLGYESFSI PVTLKVSTD NALTLHHNEN GAQYMQWQSY RTRLNTLFAR QLVARATTGI DTILSMETQN IQEPQLGKGF YATFVIPPYN LSTHGDERWF KLYIKHVVD NNSHIIYSGQ LTDTNINITL FIPLDDVPLN QDYHAKVYMT FKKSPSDGTW WGPHFVRDDK GIVTINPKSI L THFESVNV LNNISSEPMD FSGANSLYFW ELFYYTPMLV AQRLLHEQNF DEANRWLKYV WSPSGYIVHG QIQNYQWNVR PL LEDTSWN SDPLDSVDPD AVAQHDPMHY KVSTFMRTLD LLIARGDHAY RQLERDTLNE AKMWYMQALH LLGDKPYLPL STT WSDPRL DRAADITTQN AHDSAIVALR QNIPTPAPLS LRSANTLTDL FLPQINEVMM NYWQTLAQRV YNLRHNLSID GQPL YLPIY ATPADPKALL SAAVATSQGG GWLPESFMSL WRFPHMLENA RGMVSQLTQF GSTLQNIIER QDAEALNALL QNQAA ELIL TNLSIQDKTI EELDAEKTVL EKSKAGAQSR FDSYGKLYDE NINAGENQAM TLRASAAGLT TAVQASRLAG AAADLV PNI FGFAGGGSRW GAIAEATGYV MEFSANVMNT EADKISQSET YRRRRQEWEI QRNNAEAELK QIDAQLKSLA VRREAAV LQ KTSLKTQQEQ TQSQLAFLQR KFSNQALYNW LRGRLAAIYF QFYDLAVARC LMAEQAYRWE LNDDSARFIK PGAWQGTY A GLLAGETLML SLAQMEDAHL KRDKRALEVE RTVSLAEVYA GLPKDNGPFS LAQEIDKLVS QGSGSAGSGN NNLAFGAGT DTKTSLQASV SFADLKIRED YPASLGKIRR IKQISVTLPA LLGPYQDVQA ILSYGDKAGL ANGCEALAVS HGMNDSGQFQ LDFNDGKFL PFEGIAIDQG TLTLSFPNAS MPEKGKQATM LKTLNDIILH IRYTIK

UniProtKB: TcdA1

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 11.2
Component:
ConcentrationFormulaName
100.0 mMC9H19NO3SCAPS
150.0 mMNaClSodium chlorideSodium Chloride
0.1 %Tween-20
GridModel: Quantifoil R2/1 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 2 / Pretreatment - Type: GLOW DISCHARGE
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 286 K / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.5 µm / Nominal magnification: 81000
Sample stageCooling holder cryogen: NITROGEN
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Detector mode: INTEGRATING / Number grids imaged: 2 / Number real images: 24611 / Average electron dose: 57.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 936131
Startup modelType of model: PDB ENTRY
PDB model - PDB ID:
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: SPHIRE
Final 3D classificationSoftware - Name: RELION
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION
Final reconstructionApplied symmetry - Point group: C5 (5 fold cyclic) / Algorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 3.2 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION / Number images used: 13356

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Atomic model buiding 1

RefinementSpace: REAL / Protocol: FLEXIBLE FIT
Output model

PDB-8cq0:
Photorhabdus luminescens TcdA1 prepore-to-pore intermediate, K567W K2008W mutant

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