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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-12733 | |||||||||
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Title | PspA helical structure in presence of lipids | |||||||||
![]() | symmetrized map from helical refinement in CryoSparc v3.1.0 | |||||||||
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Function / homology | phage shock / PspA/IM30 / PspA/IM30 family / ![]() ![]() | |||||||||
Biological species | ![]() ![]() ![]() ![]() ![]() | |||||||||
Method | helical reconstruction / ![]() | |||||||||
![]() | Junglas B / Huber ST / Heidler T / Mann D / Henning R / Clarke M / Ortiz JO / Schneider D / Sachse C | |||||||||
![]() | ![]() Title: PspA adopts an ESCRT-III-like fold and remodels bacterial membranes. Authors: Benedikt Junglas / Stefan T Huber / Thomas Heidler / Lukas Schlösser / Daniel Mann / Raoul Hennig / Mairi Clarke / Nadja Hellmann / Dirk Schneider / Carsten Sachse / ![]() Abstract: PspA is the main effector of the phage shock protein (Psp) system and preserves the bacterial inner membrane integrity and function. Here, we present the 3.6 Å resolution cryoelectron microscopy ...PspA is the main effector of the phage shock protein (Psp) system and preserves the bacterial inner membrane integrity and function. Here, we present the 3.6 Å resolution cryoelectron microscopy (cryo-EM) structure of PspA assembled in helical rods. PspA monomers adopt a canonical ESCRT-III fold in an extended open conformation. PspA rods are capable of enclosing lipids and generating positive membrane curvature. Using cryo-EM, we visualized how PspA remodels membrane vesicles into μm-sized structures and how it mediates the formation of internalized vesicular structures. Hotspots of these activities are zones derived from PspA assemblies, serving as lipid transfer platforms and linking previously separated lipid structures. These membrane fusion and fission activities are in line with the described functional properties of bacterial PspA/IM30/LiaH proteins. Our structural and functional analyses reveal that bacterial PspA belongs to the evolutionary ancestry of ESCRT-III proteins involved in membrane remodeling. | |||||||||
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 | ![]() | 42.3 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 19.6 KB 19.6 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 12.2 KB | Display | ![]() |
Images | ![]() | 89.8 KB | ||
Masks | ![]() | 163.6 MB | ![]() | |
Others | ![]() ![]() ![]() | 42.9 MB 150.7 MB 150.7 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
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Annotation | symmetrized map from helical refinement in CryoSparc v3.1.0 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.362 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
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Projections & Slices |
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Density Histograms |
-Additional map: symmetrized and sharpened map from helical refinement in...
File | emd_12733_additional_1.map | ||||||||||||
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Annotation | symmetrized and sharpened map from helical refinement in CryoSparc v3.1.0 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: half map A
File | emd_12733_half_map_1.map | ||||||||||||
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Annotation | half map A | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: half map B
File | emd_12733_half_map_2.map | ||||||||||||
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Annotation | half map B | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : PspA helical structure in presence of lipids
Entire | Name: PspA helical structure in presence of lipids |
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Components |
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-Supramolecule #1: PspA helical structure in presence of lipids
Supramolecule | Name: PspA helical structure in presence of lipids / type: organelle_or_cellular_component / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() ![]() ![]() |
Molecular weight | Experimental: 25 KDa |
Recombinant expression | Organism: ![]() ![]() ![]() |
-Macromolecule #1: Chloroplast membrane-associated 30 kD protein
Macromolecule | Name: Chloroplast membrane-associated 30 kD protein / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() Strain: PCC 6803 / Kazusa |
Recombinant expression | Organism: ![]() ![]() ![]() |
Sequence | String: MGHHHHHHHH HSSGHIDDDD KHMELFNRVG RVLKSQLTHW QQQQEAPEDL LERLLGEMEL ELIELRRALA QTIATFKSTE RQRDAQQLI AQRWYEKAQA ALDRGNEQLA REALGQRQSY QSHTEALGKS LGEQRALVEQ VRGQLQKLER KYLELKSQKN L YLARLKSA ...String: MGHHHHHHHH HSSGHIDDDD KHMELFNRVG RVLKSQLTHW QQQQEAPEDL LERLLGEMEL ELIELRRALA QTIATFKSTE RQRDAQQLI AQRWYEKAQA ALDRGNEQLA REALGQRQSY QSHTEALGKS LGEQRALVEQ VRGQLQKLER KYLELKSQKN L YLARLKSA IAAQKIEEIA GNLDNASASS LFERIETKIL ELEAERELLN PPPSPLDKKF EQWEEQQAVE ATLAAMKARR SL PPPSS |
-Experimental details
-Structure determination
Method | ![]() |
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![]() | helical reconstruction |
Aggregation state | filament |
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Sample preparation
Concentration | 0.8 mg/mL | |||||||||
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Buffer | pH: 7.6 Component:
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Grid | Model: PELCO Ultrathin Carbon with Lacey Carbon / Material: COPPER / Mesh: 200 / Pretreatment - Type: GLOW DISCHARGE / Details: Pelco easiGlow | |||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 283.15 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | TFS TALOS |
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Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | C2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD![]() |
Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Number real images: 1531 / Average exposure time: 3.0 sec. / Average electron dose: 31.0 e/Å2 |
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Image processing
-Atomic model buiding 1
Refinement | Protocol: AB INITIO MODEL |
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