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- EMDB-52810: Salmonella typhimurium polynucleotide phosphorylase in complex wi... -
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Open data
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Basic information
Entry | ![]() | |||||||||
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Title | Salmonella typhimurium polynucleotide phosphorylase in complex with recognition site of RNase E | |||||||||
![]() | Sharpened map Salmonella PNPase in complex with RNase E | |||||||||
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![]() | polynucleotide phosphorylase / ribonuclease E / RNA degradosome / RNA BINDING PROTEIN | |||||||||
Function / homology | ![]() ribonuclease E / ribonuclease E activity / polyribonucleotide nucleotidyltransferase / polyribonucleotide nucleotidyltransferase activity / RNA catabolic process / tRNA processing / mRNA catabolic process / RNA processing / RNA endonuclease activity / cytoplasmic side of plasma membrane ...ribonuclease E / ribonuclease E activity / polyribonucleotide nucleotidyltransferase / polyribonucleotide nucleotidyltransferase activity / RNA catabolic process / tRNA processing / mRNA catabolic process / RNA processing / RNA endonuclease activity / cytoplasmic side of plasma membrane / rRNA processing / 3'-5'-RNA exonuclease activity / tRNA binding / rRNA binding / magnesium ion binding / RNA binding / zinc ion binding / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.5 Å | |||||||||
![]() | Paris G / Luisi BF | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Salmonella typhimurium polynucleotide phosphorylase in complex with recognition site of RNase E Authors: Paris G / Luisi BF | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 89.5 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 20.1 KB 20.1 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 13.4 KB | Display | ![]() |
Images | ![]() | 52.2 KB | ||
Filedesc metadata | ![]() | 6.8 KB | ||
Others | ![]() ![]() | 165.3 MB 165.3 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 1.2 MB | Display | ![]() |
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Full document | ![]() | 1.2 MB | Display | |
Data in XML | ![]() | 20.8 KB | Display | |
Data in CIF | ![]() | 26.8 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 9ibhMC 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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Annotation | Sharpened map Salmonella PNPase in complex with RNase E | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.73 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: Half map Salmonella PNPase in complex with RNase E
File | emd_52810_half_map_1.map | ||||||||||||
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Annotation | Half map Salmonella PNPase in complex with RNase E | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half map Salmonella PNPase in complex with RNase E
File | emd_52810_half_map_2.map | ||||||||||||
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Annotation | Half map Salmonella PNPase in complex with RNase E | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Polynucleotide phosphorylase in complex with recognition site fro...
Entire | Name: Polynucleotide phosphorylase in complex with recognition site from ribonuclease E |
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Components |
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-Supramolecule #1: Polynucleotide phosphorylase in complex with recognition site fro...
Supramolecule | Name: Polynucleotide phosphorylase in complex with recognition site from ribonuclease E type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#2 Details: Complex prepared by co-expression and chromatographic purification |
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Source (natural) | Organism: ![]() |
-Macromolecule #1: Polyribonucleotide nucleotidyltransferase
Macromolecule | Name: Polyribonucleotide nucleotidyltransferase / type: protein_or_peptide / ID: 1 / Details: Core of the enzyme without KH and S1 domains / Number of copies: 3 / Enantiomer: LEVO / EC number: polyribonucleotide nucleotidyltransferase |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 59.233332 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MLNPIVRKFQ YGQHTVTLET GMMARQATAA VMVSMDDTAV FVTVVGQKKA KPGQDFFPLT VNYQERTYAA GRIPGSFFRR EGRPSEGET LIARLIDRPV RPLFPEGFVN EVQVIATVVS VNPQVNPDIV AMIGASAALS LSGIPFNGPI GAARVGYIND Q YVLNPTQD ...String: MLNPIVRKFQ YGQHTVTLET GMMARQATAA VMVSMDDTAV FVTVVGQKKA KPGQDFFPLT VNYQERTYAA GRIPGSFFRR EGRPSEGET LIARLIDRPV RPLFPEGFVN EVQVIATVVS VNPQVNPDIV AMIGASAALS LSGIPFNGPI GAARVGYIND Q YVLNPTQD ELKESKLDLV VAGTEAAVLM VESEAELLSE DTMLGAVVFG HEQQQVVIQA INDLVKEAGK PRWDWQPEAV ND ALNARVA ALAESRLSDA YRITDKQERY AQVDVIKSET IEQLIAEDET LDANELGEIL HAIEKNVVRS RVLAGEPRID GRE KDMIRG LDVRTGVLPR THGSALFTRG ETQALVTATL GTARDAQVLD ELMGERTDSF LFHYNFPPYS VGETGMVGSP KRRE IGHGR LAKRGVLAVM PDMDKFPYTV RVVSEITESN GSSSMASVCG ASLALMDAGV PIKAAVAGIA MGLVKEGDNY VVLSD ILGD EDHLGDMDFK VAGSRDGISA LQMDIKIEGI TKEIMQVALN QAKGARLHIL GVMEQAINAP RG UniProtKB: Polyribonucleotide nucleotidyltransferase |
-Macromolecule #2: Ribonuclease E
Macromolecule | Name: Ribonuclease E / type: protein_or_peptide / ID: 2 / Details: C-terminal segment of ribonuclease E / Number of copies: 1 / Enantiomer: LEVO / EC number: ribonuclease E |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 6.232763 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: NHASAPMTRA PAPEYVPETP HHSDWQRPSF HFEGKGAAGG HSATRHASAP ATRPQPVE UniProtKB: Ribonuclease E |
-Macromolecule #3: water
Macromolecule | Name: water / type: ligand / ID: 3 / Number of copies: 47 / Formula: HOH |
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Molecular weight | Theoretical: 18.015 Da |
Chemical component information | ![]() ChemComp-HOH: |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 3.6 mg/mL |
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Buffer | pH: 8 Details: 20 mM Tris-HCl pH 8.0, 25 mM MgCl2, 150 mM KCl, 1 mM TCEP |
Grid | Model: UltrAuFoil R2/2 / Material: GOLD / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 60 sec. / Pretreatment - Atmosphere: AIR / Pretreatment - Pressure: 39.0 kPa / Details: 25 mA |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 277.2 K / Instrument: FEI VITROBOT MARK IV / Details: blotting force -4, 3 sec blot time. |
Details | Purified PNPase core and RNE-muGFP-CHis proteins were mixed in 1:2 ratio and their complex were separated on the Superdex 200 Increase 10/300 GL column (Cytiva) equilibrated with Cryo-EM buffer (20 mM Tris-HCl pH 8.0, 25 mM MgCl2, 150 mM KCl, 1 mM TCEP). Peak fractions were combined, the protein was concentrated to 15 microM using Amicon Ultra concentrator with 10 kDa cut-off (Millipore) and used to prepare Cryo-EM grids. The samples were mixed with CHAPSO (3-([3-cholamidopropyl]dimethylammonio)-2-hydroxy-1-propanesulfonate) at a final concentration of 8 mM |
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Electron microscopy
Microscope | TFS KRIOS |
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Specialist optics | Energy filter - Name: TFS Selectris |
Image recording | Film or detector model: FEI FALCON IV (4k x 4k) / Number grids imaged: 1 / Number real images: 12147 / Average exposure time: 4.39 sec. / Average electron dose: 50.47 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 1.8 µm / Nominal defocus min: 0.6 µm |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Initial model | Chain - Source name: AlphaFold / Chain - Initial model type: in silico model |
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Details | phenix refine and manual rebuilding using COOT |
Refinement | Space: REAL / Protocol: AB INITIO MODEL |
Output model | ![]() PDB-9ibh: |