+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-35344 | |||||||||||||||||||||||||||||||||||||||||||||||||||
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Title | Cryo-EM structure of KpFtsZ single filament | |||||||||||||||||||||||||||||||||||||||||||||||||||
Map data | ||||||||||||||||||||||||||||||||||||||||||||||||||||
Sample |
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Keywords | bacterial cell division / divisome / monobody / CELL CYCLE | |||||||||||||||||||||||||||||||||||||||||||||||||||
Function / homology | Function and homology information division septum assembly / FtsZ-dependent cytokinesis / cell division site / protein polymerization / GTPase activity / GTP binding / cytoplasm Similarity search - Function | |||||||||||||||||||||||||||||||||||||||||||||||||||
Biological species | Klebsiella pneumoniae (bacteria) | |||||||||||||||||||||||||||||||||||||||||||||||||||
Method | helical reconstruction / cryo EM / Resolution: 3.03 Å | |||||||||||||||||||||||||||||||||||||||||||||||||||
Authors | Fujita J / Amesaka H / Yoshizawa T / Kuroda N / Kamimura N / Hibino K / Konishi T / Kato Y / Hara M / Inoue T ...Fujita J / Amesaka H / Yoshizawa T / Kuroda N / Kamimura N / Hibino K / Konishi T / Kato Y / Hara M / Inoue T / Namba K / Tanaka S / Matsumura H | |||||||||||||||||||||||||||||||||||||||||||||||||||
Funding support | Japan, 16 items
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Citation | Journal: Nat Commun / Year: 2023 Title: Structures of a FtsZ single protofilament and a double-helical tube in complex with a monobody. Authors: Junso Fujita / Hiroshi Amesaka / Takuya Yoshizawa / Kota Hibino / Natsuki Kamimura / Natsuko Kuroda / Takamoto Konishi / Yuki Kato / Mizuho Hara / Tsuyoshi Inoue / Keiichi Namba / Shun-Ichi ...Authors: Junso Fujita / Hiroshi Amesaka / Takuya Yoshizawa / Kota Hibino / Natsuki Kamimura / Natsuko Kuroda / Takamoto Konishi / Yuki Kato / Mizuho Hara / Tsuyoshi Inoue / Keiichi Namba / Shun-Ichi Tanaka / Hiroyoshi Matsumura / Abstract: FtsZ polymerizes into protofilaments to form the Z-ring that acts as a scaffold for accessory proteins during cell division. Structures of FtsZ have been previously solved, but detailed mechanistic ...FtsZ polymerizes into protofilaments to form the Z-ring that acts as a scaffold for accessory proteins during cell division. Structures of FtsZ have been previously solved, but detailed mechanistic insights are lacking. Here, we determine the cryoEM structure of a single protofilament of FtsZ from Klebsiella pneumoniae (KpFtsZ) in a polymerization-preferred conformation. We also develop a monobody (Mb) that binds to KpFtsZ and FtsZ from Escherichia coli without affecting their GTPase activity. Crystal structures of the FtsZ-Mb complexes reveal the Mb binding mode, while addition of Mb in vivo inhibits cell division. A cryoEM structure of a double-helical tube of KpFtsZ-Mb at 2.7 Å resolution shows two parallel protofilaments. Our present study highlights the physiological roles of the conformational changes of FtsZ in treadmilling that regulate cell division. | |||||||||||||||||||||||||||||||||||||||||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_35344.map.gz | 97.3 MB | EMDB map data format | |
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Header (meta data) | emd-35344-v30.xml emd-35344.xml | 22.3 KB 22.3 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_35344_fsc.xml | 9.9 KB | Display | FSC data file |
Images | emd_35344.png | 22.7 KB | ||
Masks | emd_35344_msk_1.map | 103 MB | Mask map | |
Filedesc metadata | emd-35344.cif.gz | 6.8 KB | ||
Others | emd_35344_half_map_1.map.gz emd_35344_half_map_2.map.gz | 95.5 MB 95.5 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-35344 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-35344 | HTTPS FTP |
-Validation report
Summary document | emd_35344_validation.pdf.gz | 991.5 KB | Display | EMDB validaton report |
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Full document | emd_35344_full_validation.pdf.gz | 991 KB | Display | |
Data in XML | emd_35344_validation.xml.gz | 18.3 KB | Display | |
Data in CIF | emd_35344_validation.cif.gz | 23.6 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-35344 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-35344 | HTTPS FTP |
-Related structure data
Related structure data | 8ibnMC 8gzvC 8gzwC 8gzxC 8gzyC 8h1oC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_35344.map.gz / Format: CCP4 / Size: 103 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.0344 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_35344_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_35344_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_35344_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
-Entire : Cryo-EM structure of KpFtsZ single filament
Entire | Name: Cryo-EM structure of KpFtsZ single filament |
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Components |
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-Supramolecule #1: Cryo-EM structure of KpFtsZ single filament
Supramolecule | Name: Cryo-EM structure of KpFtsZ single filament / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1 |
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-Supramolecule #2: KpFtsZ
Supramolecule | Name: KpFtsZ / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1 |
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Source (natural) | Organism: Klebsiella pneumoniae (bacteria) |
-Macromolecule #1: Cell division protein FtsZ
Macromolecule | Name: Cell division protein FtsZ / type: protein_or_peptide / ID: 1 / Number of copies: 4 / Enantiomer: LEVO |
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Source (natural) | Organism: Klebsiella pneumoniae (bacteria) |
Molecular weight | Theoretical: 40.574926 KDa |
Recombinant expression | Organism: Escherichia coli (E. coli) |
Sequence | String: GHMFEPMELT NDAVIKVIGV GGGGGNAVEH MVRERIEGVE FFAVNTDAQA LRKTAVGQTI QIGSGITKGL GAGANPEVGR NAADEDREA LRAALDGADM VFIAAGMGGG TGTGAAPVVA EVAKDLGILT VAVVTKPFNF EGKKRMAFAE QGITELSKHV D SLITIPND ...String: GHMFEPMELT NDAVIKVIGV GGGGGNAVEH MVRERIEGVE FFAVNTDAQA LRKTAVGQTI QIGSGITKGL GAGANPEVGR NAADEDREA LRAALDGADM VFIAAGMGGG TGTGAAPVVA EVAKDLGILT VAVVTKPFNF EGKKRMAFAE QGITELSKHV D SLITIPND KLLKVLGRGI SLLDAFGAAN DVLKGAVQGI AELITRPGLM NVDFADVRTV MSEMGYAMMG SGVASGEDRA EE AAEMAIS SPLLEDIDLS GARGVLVNIT AGFDLRLDEF ETVGNTIRAF ASDNATVVIG TSLDPDMNDE LRVTVVATGI GMD KRPEIT LVTNKQVQQP VMDRYQQHGM SPLTQEQKPA AKVVNDNTPQ TAKEPDYLDI PAFLRKQAD UniProtKB: Cell division protein FtsZ |
-Macromolecule #2: PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER
Macromolecule | Name: PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER / type: ligand / ID: 2 / Number of copies: 4 / Formula: G2P |
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Molecular weight | Theoretical: 521.208 Da |
Chemical component information | ChemComp-G2P: |
-Macromolecule #3: POTASSIUM ION
Macromolecule | Name: POTASSIUM ION / type: ligand / ID: 3 / Number of copies: 4 / Formula: K |
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Molecular weight | Theoretical: 39.098 Da |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | helical reconstruction |
Aggregation state | filament |
-Sample preparation
Concentration | 0.5 mg/mL | |||||||||||||||
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Buffer | pH: 7.5 Component:
Details: 1 mM GMPCPP was supplemented. | |||||||||||||||
Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 200 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 20 sec. / Pretreatment - Atmosphere: AIR / Details: 20 mA | |||||||||||||||
Vitrification | Cryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | JEOL CRYO ARM 300 |
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Specialist optics | Energy filter - Name: In-column Omega Filter / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 1 / Average exposure time: 2.9 sec. / Average electron dose: 60.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.5 µm / Nominal magnification: 60000 |
Sample stage | Specimen holder model: JEOL CRYOSPECPORTER / Cooling holder cryogen: NITROGEN |