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Open data
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Basic information
Entry | ![]() | ||||||||||||
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Title | P. polymyxa GS(12)-Q-GlnR peptide | ||||||||||||
![]() | Sharpened (B factor 69.1 A^2) | ||||||||||||
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![]() | glutamine synthetase repressor dodecamer / BIOSYNTHETIC PROTEIN / LIGASE | ||||||||||||
Function / homology | ![]() polyamine catabolic process / glutamine synthetase / glutamine biosynthetic process / glutamine synthetase activity / ATP binding / metal ion binding / cytoplasm Similarity search - Function | ||||||||||||
Biological species | ![]() | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.07 Å | ||||||||||||
![]() | Travis BA / Peck J | ||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Molecular dissection of the glutamine synthetase-GlnR nitrogen regulatory circuitry in Gram-positive bacteria. Authors: Brady A Travis / Jared V Peck / Raul Salinas / Brandon Dopkins / Nicholas Lent / Viet D Nguyen / Mario J Borgnia / Richard G Brennan / Maria A Schumacher / ![]() Abstract: How bacteria sense and respond to nitrogen levels are central questions in microbial physiology. In Gram-positive bacteria, nitrogen homeostasis is controlled by an operon encoding glutamine ...How bacteria sense and respond to nitrogen levels are central questions in microbial physiology. In Gram-positive bacteria, nitrogen homeostasis is controlled by an operon encoding glutamine synthetase (GS), a dodecameric machine that assimilates ammonium into glutamine, and the GlnR repressor. GlnR detects nitrogen excess indirectly by binding glutamine-feedback-inhibited-GS (FBI-GS), which activates its transcription-repression function. The molecular mechanisms behind this regulatory circuitry, however, are unknown. Here we describe biochemical and structural analyses of GS and FBI-GS-GlnR complexes from pathogenic and non-pathogenic Gram-positive bacteria. The structures show FBI-GS binds the GlnR C-terminal domain within its active-site cavity, juxtaposing two GlnR monomers to form a DNA-binding-competent GlnR dimer. The FBI-GS-GlnR interaction stabilizes the inactive GS conformation. Strikingly, this interaction also favors a remarkable dodecamer to tetradecamer transition in some GS, breaking the paradigm that all bacterial GS are dodecamers. These data thus unveil unique structural mechanisms of transcription and enzymatic regulation. | ||||||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 306.9 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 14.7 KB 14.7 KB | Display Display | ![]() |
Images | ![]() | 115.4 KB | ||
Filedesc metadata | ![]() | 5.6 KB | ||
Others | ![]() | 163.5 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7tfaMC ![]() 7tdpC ![]() 7tdvC ![]() 7teaC ![]() 7tecC ![]() 7tenC ![]() 7tf6C ![]() 7tf7C ![]() 7tf9C ![]() 7tfbC ![]() 7tfcC ![]() 7tfdC ![]() 7tfeC 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 (B factor 69.1 A^2) | ||||||||||||||||||||
Voxel size | X=Y=Z: 0.88 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Additional map: Unsharpened
File | emd_25867_additional_1.map | ||||||||||||
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Annotation | Unsharpened | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Dodecameric P. polymyxa GS complex with glutamine and GlnR C-tail...
Entire | Name: Dodecameric P. polymyxa GS complex with glutamine and GlnR C-tail peptides |
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Components |
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-Supramolecule #1: Dodecameric P. polymyxa GS complex with glutamine and GlnR C-tail...
Supramolecule | Name: Dodecameric P. polymyxa GS complex with glutamine and GlnR C-tail peptides type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#2 |
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-Macromolecule #1: GlnR C-tail peptide
Macromolecule | Name: GlnR C-tail peptide / type: protein_or_peptide / ID: 1 / Number of copies: 12 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 1.210402 KDa |
Sequence | String: LIQGELSRFF |
-Macromolecule #2: Glutamine synthetase
Macromolecule | Name: Glutamine synthetase / type: protein_or_peptide / ID: 2 / Number of copies: 12 / Enantiomer: LEVO / EC number: glutamine synthetase |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 52.51034 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MGSSHHHHHH SSGLVPRGSH MSYTREDIIR IAEEENVRFI RLQFTDLLGT IKNVEIPVSQ LEKALDNKMM FDGSSIEGYV RIEESDMYL YPDLDTWVVF PWVTSDRVAR LICDIYKPDG SPFAGDPRGI LKRVLKEAEE LGYTSMNVGP EPEFFLFKTD E KGDPTTEL ...String: MGSSHHHHHH SSGLVPRGSH MSYTREDIIR IAEEENVRFI RLQFTDLLGT IKNVEIPVSQ LEKALDNKMM FDGSSIEGYV RIEESDMYL YPDLDTWVVF PWVTSDRVAR LICDIYKPDG SPFAGDPRGI LKRVLKEAEE LGYTSMNVGP EPEFFLFKTD E KGDPTTEL NDQGGYFDLA PMDLGENCRR EIVLKLEEMG FEIEASHHEV APGQHEIDFK YADAVKAADQ IQTFKLVVKT IA RQHGLHA TFMPKPLFGV NGSGMHCNQS LFKDNENVFY DETDELGLSQ TARHYMAGIL KHARAMAAIT NPTVNSYKRL VPG YEAPCY VAWSASNRSP MIRIPASRGL STRVEVRNPD PAANPYLALA VMLRAGLDGI KRQMALPAPI DRNIYVMSEE ERIE EGIPS LPADLKEALS ELIRSEVISD ALGDHALAYF YELKEIEWDM YRTQVHQWER DQYLTLY UniProtKB: Glutamine synthetase |
-Macromolecule #3: MAGNESIUM ION
Macromolecule | Name: MAGNESIUM ION / type: ligand / ID: 3 / Number of copies: 24 / Formula: MG |
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Molecular weight | Theoretical: 24.305 Da |
-Macromolecule #4: GLUTAMINE
Macromolecule | Name: GLUTAMINE / type: ligand / ID: 4 / Number of copies: 12 / Formula: GLN |
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Molecular weight | Theoretical: 146.144 Da |
Chemical component information | ![]() ChemComp-GLN: |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 0.8 mg/mL |
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Buffer | pH: 7.5 |
Vitrification | Cryogen name: ETHANE |
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Electron microscopy
Microscope | FEI TALOS ARCTICA |
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Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 46.27 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.1 µm / Nominal defocus min: 0.5 µm |
Experimental equipment | ![]() Model: Talos Arctica / Image courtesy: FEI Company |
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Image processing
Startup model | Type of model: NONE |
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Final reconstruction | Applied symmetry - Point group: D6 (2x6 fold dihedral) / Resolution.type: BY AUTHOR / Resolution: 2.07 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 3.2) / Number images used: 1743527 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 3.2) |
Final angle assignment | Type: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 3.2) |
-Atomic model buiding 1
Refinement | Space: REAL |
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Output model | ![]() PDB-7tfa: |