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- PDB-6sfw: Cryo-EM Structure of the ClpX component of the ClpXP1/2 degradati... -
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Basic information
Entry | Database: PDB / ID: 6sfw | ||||||||||||
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Title | Cryo-EM Structure of the ClpX component of the ClpXP1/2 degradation machinery. | ||||||||||||
![]() | ATP-dependent Clp protease ATP-binding subunit ClpX | ||||||||||||
![]() | TRANSPORT PROTEIN / AAA+ / ATPase / Protease / Listeria / Motor protein / chaperone | ||||||||||||
Function / homology | ![]() protein catabolic process / unfolded protein binding / protein folding / peptidase activity / protein dimerization activity / cell division / ATP hydrolysis activity / proteolysis / zinc ion binding / ATP binding Similarity search - Function | ||||||||||||
Biological species | ![]() | ||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 6 Å | ||||||||||||
![]() | Gatsogiannis, C. / Merino, F. / Raunser, S. | ||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Cryo-EM structure of the ClpXP protein degradation machinery. Authors: Christos Gatsogiannis / Dora Balogh / Felipe Merino / Stephan A Sieber / Stefan Raunser / ![]() Abstract: The ClpXP machinery is a two-component protease complex that performs targeted protein degradation in bacteria and mitochondria. The complex consists of the AAA+ chaperone ClpX and the peptidase ClpP. ...The ClpXP machinery is a two-component protease complex that performs targeted protein degradation in bacteria and mitochondria. The complex consists of the AAA+ chaperone ClpX and the peptidase ClpP. The hexameric ClpX utilizes the energy of ATP binding and hydrolysis to engage, unfold and translocate substrates into the catalytic chamber of tetradecameric ClpP, where they are degraded. Formation of the complex involves a symmetry mismatch, because hexameric AAA+ rings bind axially to the opposing stacked heptameric rings of the tetradecameric ClpP. Here we present the cryo-EM structure of ClpXP from Listeria monocytogenes. We unravel the heptamer-hexamer binding interface and provide novel insight into the ClpX-ClpP cross-talk and activation mechanism. Comparison with available crystal structures of ClpP and ClpX in different states allows us to understand important aspects of the complex mode of action of ClpXP and provides a structural framework for future pharmacological applications. | ||||||||||||
History |
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Structure visualization
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Structure viewer | Molecule: ![]() ![]() |
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Downloads & links
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PDBx/mmCIF format | ![]() | 339.8 KB | Display | ![]() |
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PDB format | ![]() | 278.8 KB | Display | ![]() |
PDBx/mmJSON format | ![]() | Tree view | ![]() | |
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-Validation report
Summary document | ![]() | 1 MB | Display | ![]() |
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Full document | ![]() | 1.1 MB | Display | |
Data in XML | ![]() | 67.5 KB | Display | |
Data in CIF | ![]() | 98.2 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 10162MC ![]() 6sfxC M: map data used to model this data C: citing same article ( |
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Similar structure data |
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Links
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Assembly
Deposited unit | ![]()
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Components
#1: Protein | Mass: 46492.172 Da / Num. of mol.: 6 / Mutation: E183Q Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: clpX, ARJ20_08095, B4Y57_02215, D3B94_07125, DWE46_10580, DWE48_07885, FA029_07915, FORC68_1290, LmNIHS28_00678 Production host: ![]() ![]() |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
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Sample preparation
Component | Name: LmClpX / Type: COMPLEX Details: Hexameric AAA+ ATPase that unfolds the substrate to be degraded by ClpP1/2 Entity ID: all / Source: RECOMBINANT | ||||||||||||||||||||||||||||
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Molecular weight | Value: 1.7 / Experimental value: NO | ||||||||||||||||||||||||||||
Source (natural) | Organism: ![]() | ||||||||||||||||||||||||||||
Source (recombinant) | Organism: ![]() ![]() | ||||||||||||||||||||||||||||
Buffer solution | pH: 7.6 Details: The sample was crosslinked with 0.1% glutaraldehyde. The reaction was quenched after 30 s with 2 eq Tris-HCl. | ||||||||||||||||||||||||||||
Buffer component |
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Specimen | Conc.: 0.01 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | ||||||||||||||||||||||||||||
Specimen support | Grid material: COPPER / Grid mesh size: 400 divisions/in. / Grid type: Quantifoil R2/2 | ||||||||||||||||||||||||||||
Vitrification | Instrument: GATAN CRYOPLUNGE 3 / Cryogen name: ETHANE / Humidity: 90 % Details: The sample was blotted after 45 s of incubation time |
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Electron microscopy imaging
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: ![]() |
Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 59000 X / Calibrated magnification: 112807 X / Cs: 0 mm |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Average exposure time: 2 sec. / Electron dose: 114 e/Å2 / Detector mode: INTEGRATING / Film or detector model: FEI FALCON III (4k x 4k) / Num. of grids imaged: 1 / Num. of real images: 3200 Details: Images were collected in movie mode at a frame rate of 50 ms |
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Processing
EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Particle selection | Num. of particles selected: 613322 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
3D reconstruction | Resolution: 6 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 383927 Details: The overall resolution of the map is 4 A, but the ClpX region has significantly lower local resolution. Symmetry type: POINT | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Atomic model building | Protocol: FLEXIBLE FIT / Space: REAL |