+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-4568 | |||||||||
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Title | Cryo-EM 3D structure of CI2eng toroidal dodecameric assembly. | |||||||||
Map data | 3D reconstruction of CI2eng assembly formed in solution. Resol 8.55A | |||||||||
Sample |
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Biological species | Hordeum vulgare (barley) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 8.55 Å | |||||||||
Authors | Valpuesta JM / Alvira S / Campos LA / Munoz V | |||||||||
Funding support | Spain, 2 items
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Citation | Journal: Nat Commun / Year: 2019 Title: Engineering protein assemblies with allosteric control via monomer fold-switching. Authors: Luis A Campos / Rajendra Sharma / Sara Alvira / Federico M Ruiz / Beatriz Ibarra-Molero / Mourad Sadqi / Carlos Alfonso / Germán Rivas / Jose M Sanchez-Ruiz / Antonio Romero Garrido / José ...Authors: Luis A Campos / Rajendra Sharma / Sara Alvira / Federico M Ruiz / Beatriz Ibarra-Molero / Mourad Sadqi / Carlos Alfonso / Germán Rivas / Jose M Sanchez-Ruiz / Antonio Romero Garrido / José M Valpuesta / Victor Muñoz / Abstract: The macromolecular machines of life use allosteric control to self-assemble, dissociate and change shape in response to signals. Despite enormous interest, the design of nanoscale allosteric ...The macromolecular machines of life use allosteric control to self-assemble, dissociate and change shape in response to signals. Despite enormous interest, the design of nanoscale allosteric assemblies has proven tremendously challenging. Here we present a proof of concept of allosteric assembly in which an engineered fold switch on the protein monomer triggers or blocks assembly. Our design is based on the hyper-stable, naturally monomeric protein CI2, a paradigm of simple two-state folding, and the toroidal arrangement with 6-fold symmetry that it only adopts in crystalline form. We engineer CI2 to enable a switch between the native and an alternate, latent fold that self-assembles onto hexagonal toroidal particles by exposing a favorable inter-monomer interface. The assembly is controlled on demand via the competing effects of temperature and a designed short peptide. These findings unveil a remarkable potential for structural metamorphosis in proteins and demonstrate key principles for engineering protein-based nanomachinery. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_4568.map.gz | 801.7 KB | EMDB map data format | |
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Header (meta data) | emd-4568-v30.xml emd-4568.xml | 9.9 KB 9.9 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_4568_fsc.xml | 2.3 KB | Display | FSC data file |
Images | emd_4568.png | 154.3 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-4568 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-4568 | HTTPS FTP |
-Validation report
Summary document | emd_4568_validation.pdf.gz | 246.9 KB | Display | EMDB validaton report |
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Full document | emd_4568_full_validation.pdf.gz | 246.1 KB | Display | |
Data in XML | emd_4568_validation.xml.gz | 6.3 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-4568 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-4568 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_4568.map.gz / Format: CCP4 / Size: 844.7 KB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | 3D reconstruction of CI2eng assembly formed in solution. Resol 8.55A | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.71 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : CI2eng modification of the chymotrypsin inhibitor 2
Entire | Name: CI2eng modification of the chymotrypsin inhibitor 2 |
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Components |
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-Supramolecule #1: CI2eng modification of the chymotrypsin inhibitor 2
Supramolecule | Name: CI2eng modification of the chymotrypsin inhibitor 2 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Hordeum vulgare (barley) / Organ: seeds |
Recombinant expression | Organism: Escherichia coli (E. coli) |
Molecular weight | Experimental: 80 KDa |
-Macromolecule #1: CI2eng - modification of the chymotrypsin inhibitor 2 (CI2)
Macromolecule | Name: CI2eng - modification of the chymotrypsin inhibitor 2 (CI2) type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Hordeum vulgare (barley) |
Recombinant expression | Organism: Escherichia coli (E. coli) |
Sequence | String: MDAKTEWPE LVGKSLEEAK KALLQDKPEA TIIVIPVGTI VTMEYRVDRV RIVVDKLDNI AEVPTVG |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 8.5 / Details: 20mM sodium borate buffer at pH 8.5 |
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Grid | Model: Quantifoil R1.2/1.3 / Mesh: 300 |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TECNAI ARCTICA |
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Image recording | Film or detector model: FEI FALCON III (4k x 4k) / Average electron dose: 2.5 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Talos Arctica / Image courtesy: FEI Company |