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Open data
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Basic information
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Title | Structure of the autoinducer-2 exporter TqsA from E. coli | |||||||||
![]() | C5 symmetry imposed final map for TqsA | |||||||||
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![]() | cryo-EM / membrane protein / autoinducer-2 exporter / quorum sensing / pentamer / protein oligomerization / STRUCTURAL PROTEIN | |||||||||
Function / homology | autoinducer AI-2 transmembrane transport / Transmembrane protein TqsA-like / AI-2E family transporter / quorum sensing / plasma membrane => GO:0005886 / efflux transmembrane transporter activity / transmembrane transport / AI-2 transport protein TqsA![]() | |||||||||
Biological species | ![]() ![]() ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.3 Å | |||||||||
![]() | Khera R / Xie H | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Cryo-EM structures of pentameric autoinducer-2 exporter from Escherichia coli reveal its transport mechanism. Authors: Radhika Khera / Ahmad R Mehdipour / Jani R Bolla / Joerg Kahnt / Sonja Welsch / Ulrich Ermler / Cornelia Muenke / Carol V Robinson / Gerhard Hummer / Hao Xie / Hartmut Michel / ![]() ![]() ![]() Abstract: Bacteria utilize small extracellular molecules to communicate in order to collectively coordinate their behaviors in response to the population density. Autoinducer-2 (AI-2), a universal molecule for ...Bacteria utilize small extracellular molecules to communicate in order to collectively coordinate their behaviors in response to the population density. Autoinducer-2 (AI-2), a universal molecule for both intra- and inter-species communication, is involved in the regulation of biofilm formation, virulence, motility, chemotaxis, and antibiotic resistance. While many studies have been devoted to understanding the biosynthesis and sensing of AI-2, very little information is available on its export. The protein TqsA from Escherichia coli, which belongs to the AI-2 exporter superfamily, has been shown to export AI-2. Here, we report the cryogenic electron microscopic structures of two AI-2 exporters (TqsA and YdiK) from E. coli at 3.35 Å and 2.80 Å resolutions, respectively. Our structures suggest that the AI-2 exporter exists as a homo-pentameric complex. In silico molecular docking and native mass spectrometry experiments were employed to demonstrate the interaction between AI-2 and TqsA, and the results highlight the functional importance of two helical hairpins in substrate binding. We propose that each monomer works as an independent functional unit utilizing an elevator-type transport mechanism. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 11.7 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 19 KB 19 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 11.4 KB | Display | ![]() |
Images | ![]() | 107 KB | ||
Masks | ![]() | 125 MB | ![]() | |
Filedesc metadata | ![]() | 5.9 KB | ||
Others | ![]() ![]() ![]() | 13.5 MB 98.3 MB 98.3 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7nb6MC ![]() 7ot9C 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 | C5 symmetry imposed final map for TqsA | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.837 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: C1 symmetry map
File | emd_12256_additional_1.map | ||||||||||||
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Annotation | C1 symmetry map | ||||||||||||
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-Half map: Half map2
File | emd_12256_half_map_1.map | ||||||||||||
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Annotation | Half map2 | ||||||||||||
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-Half map: Half map1
File | emd_12256_half_map_2.map | ||||||||||||
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Annotation | Half map1 | ||||||||||||
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Sample components
-Entire : Pentameric TqsA
Entire | Name: Pentameric TqsA |
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Components |
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-Supramolecule #1: Pentameric TqsA
Supramolecule | Name: Pentameric TqsA / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 187 KDa |
-Macromolecule #1: AI-2 transport protein TqsA
Macromolecule | Name: AI-2 transport protein TqsA / type: protein_or_peptide / ID: 1 / Number of copies: 5 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 37.563543 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MAKPIITLNG LKIVIMLGML VIILCGIRFA AEIIVPFILA LFIAVILNPL VQHMVRWRVP RVLAVSILMT IIVMAMVLLL AYLGSALNE LTRTLPQYRN SIMTPLQALE PLLQRVGIDV SVDQLAHYID PNAAMTLLTN LLTQLSNAMS SIFLLLLTVL F MLLEVPQL ...String: MAKPIITLNG LKIVIMLGML VIILCGIRFA AEIIVPFILA LFIAVILNPL VQHMVRWRVP RVLAVSILMT IIVMAMVLLL AYLGSALNE LTRTLPQYRN SIMTPLQALE PLLQRVGIDV SVDQLAHYID PNAAMTLLTN LLTQLSNAMS SIFLLLLTVL F MLLEVPQL PGKFQQMMAR PVEGMAAIQR AIDSVSHYLV LKTAISIITG LVAWAMLAAL DVRFAFVWGL LAFALNYIPN IG SVLAAIP PIAQVLVFNG FYEALLVLAG YLLINLVFGN ILEPRIMGRG LGLSTLVVFL SLIFWGWLLG PVGMLLSVPL TII VKIALE QTAGGQSIAV LLSDLNKE UniProtKB: AI-2 transport protein TqsA |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 3.4 mg/mL |
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Buffer | pH: 7.5 / Details: 50 mM Tris (pH 7.5), 150 mM NaCl and 0.006% GDN |
Grid | Model: C-flat-1.2/1.3 / Material: COPPER / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK III / Details: blot time 4 s, blot force +20. |
Details | Its a membrane protein and for purification, glyco-diosgenin was used. |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Digitization - Dimensions - Width: 5760 pixel / Digitization - Dimensions - Height: 4092 pixel / Number grids imaged: 1 / Number real images: 5452 / Average electron dose: 80.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: OTHER / Imaging mode: OTHER / Cs: 2.7 mm |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |