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Open data
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Basic information
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Title | E. coli MraY mutant-T23P | |||||||||
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Function / homology | ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() Similarity search - Function | |||||||||
Biological species | ![]() ![]() ![]() | |||||||||
Method | ![]() ![]() | |||||||||
![]() | Orta AK / Li YE / Clemons WM | |||||||||
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![]() | ![]() Title: Synthesis of lipid-linked precursors of the bacterial cell wall is governed by a feedback control mechanism in Pseudomonas aeruginosa. Authors: Lindsey S Marmont / Anna K Orta / Becca W A Baileeves / David Sychantha / Ana Fernández-Galliano / Yancheng E Li / Neil G Greene / Robin A Corey / Phillip J Stansfeld / William M Clemons / ...Authors: Lindsey S Marmont / Anna K Orta / Becca W A Baileeves / David Sychantha / Ana Fernández-Galliano / Yancheng E Li / Neil G Greene / Robin A Corey / Phillip J Stansfeld / William M Clemons / Thomas G Bernhardt / ![]() ![]() ![]() Abstract: Many bacterial surface glycans such as the peptidoglycan (PG) cell wall are built from monomeric units linked to a polyprenyl lipid carrier. How this limiting carrier is distributed among competing ...Many bacterial surface glycans such as the peptidoglycan (PG) cell wall are built from monomeric units linked to a polyprenyl lipid carrier. How this limiting carrier is distributed among competing pathways has remained unclear. Here we describe the isolation of hyperactive variants of Pseudomonas aeruginosa MraY, the enzyme that forms the first lipid-linked PG precursor. These variants result in the elevated production of the final PG precursor lipid II in cells and are hyperactive in vitro. The activated MraY variants have substitutions that map to a cavity on the extracellular side of the dimer interface, far from the active site. Our structural and molecular dynamics results suggest that this cavity is a binding site for externalized lipid II. Overall, our results support a model in which excess externalized lipid II allosterically inhibits MraY, providing a feedback mechanism that prevents the sequestration of lipid carrier in the PG biogenesis pathway. | |||||||||
History |
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Structure visualization
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Downloads & links
-EMDB archive
Map data | ![]() | 48.6 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 19.2 KB 19.2 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 11.1 KB | Display | ![]() |
Images | ![]() | 103.2 KB | ||
Masks | ![]() | 52.7 MB | ![]() | |
Filedesc metadata | ![]() | 6.1 KB | ||
Others | ![]() ![]() ![]() | 49.6 MB 49 MB 49 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8tluMC 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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Map
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Annotation | sharpened map | ||||||||||||||||||||
Voxel size | X=Y=Z: 0.832 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
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-Additional map: unsharpened map
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-Half map: #2
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-Half map: #1
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Sample components
-Entire : E. coli MraY T23P dimer
Entire | Name: E. coli MraY T23P dimer |
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Components |
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-Supramolecule #1: E. coli MraY T23P dimer
Supramolecule | Name: E. coli MraY T23P dimer / type: organelle_or_cellular_component / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() ![]() ![]() |
Molecular weight | Theoretical: 39.875 KDa |
-Macromolecule #1: Phospho-N-acetylmuramoyl-pentapeptide-transferase
Macromolecule | Name: Phospho-N-acetylmuramoyl-pentapeptide-transferase / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() ![]() |
Molecular weight | Theoretical: 39.905551 KDa |
Recombinant expression | Organism: ![]() ![]() ![]() |
Sequence | String: MLVWLAEHLV KYYSGFNVFS YLPFRAIVSL LTALFISLWM GPRMIAHLQK LSFGQVVRND GPESHFSKRG TPTMGGIMIL TAIVISVLL WAYPSNPYVW CVLVVLVGYG VIGFVDDYRK VVRKDTKGLI ARWKYFWMSV IALGVAFALY LAGKDTPATQ L VVPFFKDV ...String: MLVWLAEHLV KYYSGFNVFS YLPFRAIVSL LTALFISLWM GPRMIAHLQK LSFGQVVRND GPESHFSKRG TPTMGGIMIL TAIVISVLL WAYPSNPYVW CVLVVLVGYG VIGFVDDYRK VVRKDTKGLI ARWKYFWMSV IALGVAFALY LAGKDTPATQ L VVPFFKDV MPQLGLFYIL LAYFVIVGTG NAVNLTDGLD GLAIMPTVFV AGGFALVAWA TGNMNFASYL HIPYLRHAGE LV IVCTAIV GAGLGFLWFN TYPAQVFMGD VGSLALGGAL GIIAVLLRQE FLLVIMGGVF VVETLSVILQ VGSFKLRGQR IFR MAPIHH HYELKGWPEP RVIVRFWIIS LMLVLIGLAT LKVR UniProtKB: ![]() |
-Experimental details
-Structure determination
Method | ![]() |
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Aggregation state | particle |
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Sample preparation
Concentration | 5 mg/mL |
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Buffer | pH: 7.5 |
Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 60 sec. |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV |
Details | MraY was pulled down using the YES complex. |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: OTHER / Imaging mode: BRIGHT FIELD![]() |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 1 / Average electron dose: 60.0 e/Å2 |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |