|Entry||Database: EMDB / ID: 5330|
|Title||Cryo-electron tomography reveals novel interactions and doublet-specific structures in the I1 dynein|
|Keywords||axoneme / molecular motor / motility regulation / flagella|
|Sample||Cryo-electron tomography and subtomographic average (750 axonemal repeats) of isolated axonemes of wild type Chlamydomonas (CC 125, 137c), I1 dynein complex (dynein f) is bound to the doublet microtubule and is connected to neighboring structures.|
|Source||Chlamydomonas reinhardtii / plant / unicellular green algae|
|Map data||This is a subtomogram average of the I1 inner dynein complex in wild type Chlamydomonas flagella|
|Method||subtomogram averaging, at 39 Å resolution|
|Authors||Heuser T / Barber CF / Lin J / Krell J / Rebesco M / Porter ME / Nicastro D|
|Citation||Proc. Natl. Acad. Sci. U.S.A., 2012, 109, E2067-E2076|
Proc. Natl. Acad. Sci. U.S.A., 2012, 109, E2067-E2076 Yorodumi Papers
|Date||Deposition: Aug 10, 2011 / Header (metadata) release: Aug 11, 2011 / Map release: Jun 21, 2012 / Last update: Apr 2, 2014|
Downloads & links
|File||emd_5330.map.gz (map file in CCP4 format, 345 KB)|
|Projections & slices|
Images are generated by Spider package.
|Voxel size||X=Y=Z: 9.5 Å|
CCP4 map header:
-Entire Cryo-electron tomography and subtomographic average (750 axonemal...
|Entire||Name: Cryo-electron tomography and subtomographic average (750 axonemal repeats) of isolated axonemes of wild type Chlamydomonas (CC 125, 137c), I1 dynein complex (dynein f) is bound to the doublet microtubule and is connected to neighboring structures.|
Number of components: 1
-Component #1: cellular-component, I1 dynein complex
|Cellular-component||Name: I1 dynein complex / a.k.a: dynein f / Recombinant expression: No / Number of Copies: 2|
|Source||Species: Chlamydomonas reinhardtii / plant / unicellular green algae|
Strain: CC-125, 137c
|Source (natural)||Organelle: eukaryotic flagella / Cell: Chlamydomonas reinhardtii|
|Sample solution||Specimen conc.: 1 mg/ml|
Buffer solution: 10 mM HEPES, pH 7.4, 25 mM NaCl, 4 mM MgSO4, 1 mM EGTA, 0.1 mM EDTA
|Support film||Quantifoil holey carbon grids Cu 200 mesh R2/2|
|Vitrification||Instrument: HOMEMADE PLUNGER / Cryogen name: ETHANE / Temperature: 100 K / Method: front-side blotting for 2-3 seconds|
-Electron microscopy imaging
Model: Tecnai F30 / Image courtesy: FEI Company
|Imaging||Microscope: FEI TECNAI F30 / Date: Feb 20, 2004|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 100 e/Å2 / Illumination mode: OTHER|
|Lens||Magnification: 13500 X (nominal) / Imaging mode: BRIGHT FIELD / Defocus: 6000 - 8000 nm / Energy filter: GATAN postcolumn filter GIF / Energy window: 0-20 eV|
|Specimen Holder||Holder: Eucentric / Model: GATAN LIQUID NITROGEN / Tilt Angle: -60 - 60 deg. / Temperature: 80 K|
|Camera||Detector: GENERIC GATAN (2k x 2k)|
|Processing||Method: subtomogram averaging|
Details: 750 axonemal repeats (96 nm long) from 5 tomograms (reconstructed using fiducial alignment and weighted backprojection, IMOD software, Kremer et al. 1996) were aligned and averaged using the PEET software (bio3d.colorado.edu, Nicastro et al. 2006). Average number of tilts used in the 3D reconstructions: 80. Average tomographic tilt angle increment: 1.5.
|3D reconstruction||Algorithm: fiducial alignment and weighted backprojection / Software: IMOD|
Details: Final maps were calculated by averaging 750 particles from 5 tomograms
Resolution: 39 Å / Resolution method: FSC 0.5
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