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- EMDB-6412: Structure of PhnGI complex from Escherichia coli by negative stain -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-6412 | |||||||||
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Title | Structure of PhnGI complex from Escherichia coli by negative stain | |||||||||
![]() | PhnGI complex from Escherichia coli | |||||||||
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![]() | PhnGI | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / negative staining / Resolution: 23.0 Å | |||||||||
![]() | Yang K / Ren Z / Raushel FM / Zhang J | |||||||||
![]() | ![]() Title: Structures of the Carbon-Phosphorus Lyase Complex Reveal the Binding Mode of the NBD-like PhnK. Authors: Kailu Yang / Zhongjie Ren / Frank M Raushel / Junjie Zhang / ![]() Abstract: The carbon-phosphorus (C-P) lyase complex is essential for the metabolism of unactivated phosphonates to phosphate in bacteria. Using single-particle cryo-electron microscopy, we determined two ...The carbon-phosphorus (C-P) lyase complex is essential for the metabolism of unactivated phosphonates to phosphate in bacteria. Using single-particle cryo-electron microscopy, we determined two structures of the C-P lyase core complex PhnG2H2I2J2, with or without PhnK. PhnG2H2I2J2 is a two-fold symmetric hetero-octamer. Its two PhnJ subunits provide two identical binding sites for PhnK. Only one PhnK binds to PhnG2H2I2J2 due to steric hindrance. PhnK is homologous to the nucleotide-binding domain (NBD) of ATP-binding cassette transporters. The α helices 3 and 4 of PhnK bind to α helix 6 and a loop (residues 227-230) of PhnJ, in a different mode from the binding of NBDs to their transmembrane partners. Moreover, binding of PhnK exposes the active site residue, Gly32 of PhnJ, located near the interface between PhnJ and PhnH. This structural information provides a basis for further deciphering of the reaction mechanism of the C-P lyase. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 30.7 KB | ![]() | |
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Header (meta data) | ![]() ![]() | 9.7 KB 9.7 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 1.2 KB | Display | ![]() |
Images | ![]() | 25.4 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 79.2 KB | Display | ![]() |
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Full document | ![]() | 78.3 KB | Display | |
Data in XML | ![]() | 494 B | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | PhnGI complex from Escherichia coli | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 5.3 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : PhnGI complex from Escherichia coli
Entire | Name: PhnGI complex from Escherichia coli |
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Components |
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-Supramolecule #1000: PhnGI complex from Escherichia coli
Supramolecule | Name: PhnGI complex from Escherichia coli / type: sample / ID: 1000 / Oligomeric state: Two PhnG, two PhnI / Number unique components: 2 |
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Molecular weight | Theoretical: 115 KDa |
-Macromolecule #1: PhnG
Macromolecule | Name: PhnG / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Recombinant expression: Yes |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 18.7 KDa |
Recombinant expression | Organism: ![]() ![]() |
-Macromolecule #2: PhnI
Macromolecule | Name: PhnI / type: protein_or_peptide / ID: 2 / Number of copies: 2 / Recombinant expression: Yes |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 38.9 KDa |
Recombinant expression | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | negative staining |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 0.01 mg/mL |
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Buffer | pH: 8.5 / Details: 50 mM HEPES, 150 mM NaCl, 2 mM TCEP |
Staining | Type: NEGATIVE / Details: sample was stained by 2% uranyl acetate |
Grid | Details: 200 mesh copper grid with carbon film |
Vitrification | Cryogen name: NONE / Instrument: OTHER |
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Electron microscopy
Microscope | FEI TECNAI F20 |
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Date | May 21, 2014 |
Image recording | Category: CCD / Film or detector model: GATAN ULTRASCAN 1000 (2k x 2k) / Number real images: 20 / Average electron dose: 40 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Calibrated magnification: 26285 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2 mm / Nominal defocus max: 4.1 µm / Nominal defocus min: 3.0 µm / Nominal magnification: 40000 |
Sample stage | Specimen holder model: SIDE ENTRY, EUCENTRIC |
Experimental equipment | ![]() Model: Tecnai F20 / Image courtesy: FEI Company |