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Open data
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Basic information
Entry | ![]() | |||||||||
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Title | TadA/CpaF with ADP | |||||||||
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![]() | pilus / secretion / ATPase / MOTOR PROTEIN | |||||||||
Function / homology | : / Type II/IV secretion system protein / Type II/IV secretion system protein / P-loop containing nucleoside triphosphate hydrolase / Pilus assembly ATPase CpaF![]() | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.1 Å | |||||||||
![]() | Hohl M / Low H | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Bidirectional pilus processing in the Tad pilus system motor CpaF. Authors: Michael Hohl / Emma J Banks / Max P Manley / Tung B K Le / Harry H Low / ![]() Abstract: The bacterial tight adherence pilus system (TadPS) assembles surface pili essential for adhesion and colonisation in many human pathogens. Pilus dynamics are powered by the ATPase CpaF (TadA), which ...The bacterial tight adherence pilus system (TadPS) assembles surface pili essential for adhesion and colonisation in many human pathogens. Pilus dynamics are powered by the ATPase CpaF (TadA), which drives extension and retraction cycles in Caulobacter crescentus through an unknown mechanism. Here we use cryogenic electron microscopy and cell-based light microscopy to characterise CpaF mechanism. We show that CpaF assembles into a hexamer with C2 symmetry in different nucleotide states. Nucleotide cycling occurs through an intra-subunit clamp-like mechanism that promotes sequential conformational changes between subunits. Moreover, a comparison of the active sites with different nucleotides bound suggests a mechanism for bidirectional motion. Conserved CpaF residues, predicted to interact with platform proteins CpaG (TadB) and CpaH (TadC), are mutated in vivo to establish their role in pilus processing. Our findings provide a model for how CpaF drives TadPS pilus dynamics and have broad implications for how other ancient type 4 filament family members power pilus assembly. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 161.7 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 14.1 KB 14.1 KB | Display Display | ![]() |
Images | ![]() | 173 KB | ||
Filedesc metadata | ![]() | 5.6 KB | ||
Others | ![]() ![]() | 139.3 MB 138.9 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8rjfMC ![]() 19275 ![]() 19279 ![]() 8rkdC ![]() 8rklC M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Voxel size | X=Y=Z: 0.829 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
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Sample components
-Entire : Hexameric complex of TadA bound to ADP
Entire | Name: Hexameric complex of TadA bound to ADP |
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Components |
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-Supramolecule #1: Hexameric complex of TadA bound to ADP
Supramolecule | Name: Hexameric complex of TadA bound to ADP / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1 |
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Source (natural) | Organism: ![]() |
-Macromolecule #1: Pilus assembly ATPase CpaF
Macromolecule | Name: Pilus assembly ATPase CpaF / type: protein_or_peptide / ID: 1 / Number of copies: 6 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 46.578355 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: DYYHATKTTI FNALLNTIDL SQLAQLDLKQ AGEEIRDIVA ELVAIKNVSM SVAEQEHLVQ DIINDVLGYG PLEPLLARDD IADIMVNGA HRVFIEVGGK VQLTNVRFRD NLQLMNICQR IVSQVGRRVD ESSPICDARL PDGSRVNVIA PPLALDGPTL T IRKFKKDK ...String: DYYHATKTTI FNALLNTIDL SQLAQLDLKQ AGEEIRDIVA ELVAIKNVSM SVAEQEHLVQ DIINDVLGYG PLEPLLARDD IADIMVNGA HRVFIEVGGK VQLTNVRFRD NLQLMNICQR IVSQVGRRVD ESSPICDARL PDGSRVNVIA PPLALDGPTL T IRKFKKDK LTMKNLVEFA SISPEGARVL GVIGACRCNL VISGGTGSGK TTLLNTMTAF IDPTERVVTC EDAAELQLQQ PH VVRLETR PPNLEGSGAV TMRDLVKNCL RMRPERIIVG EVRGPEAFDL LQAMNTGHDG SMGTLHANSP REAISRIESM ITM GGYGLP SKTIKEMIVG SVDVIIQAAR LRDGSRRITH ITEVVGLEGD VIVTQDLFVY EITGEDEHGK VVGKHRSTGI ARPR FWDRA RYYGLERELA EALDAAEAL UniProtKB: Pilus assembly ATPase CpaF |
-Macromolecule #2: ADENOSINE-5'-DIPHOSPHATE
Macromolecule | Name: ADENOSINE-5'-DIPHOSPHATE / type: ligand / ID: 2 / Number of copies: 6 / Formula: ADP |
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Molecular weight | Theoretical: 427.201 Da |
Chemical component information | ![]() ChemComp-ADP: |
-Macromolecule #3: MAGNESIUM ION
Macromolecule | Name: MAGNESIUM ION / type: ligand / ID: 3 / Number of copies: 6 / Formula: MG |
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Molecular weight | Theoretical: 24.305 Da |
-Macromolecule #4: 4-(2-HYDROXYETHYL)-1-PIPERAZINE ETHANESULFONIC ACID
Macromolecule | Name: 4-(2-HYDROXYETHYL)-1-PIPERAZINE ETHANESULFONIC ACID / type: ligand / ID: 4 / Number of copies: 4 / Formula: EPE |
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Molecular weight | Theoretical: 238.305 Da |
Chemical component information | ![]() ChemComp-EPE: |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.4 |
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Vitrification | Cryogen name: ETHANE |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 40.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.0 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
Startup model | Type of model: INSILICO MODEL |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.1 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 114641 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |