|Entry||Database: EMDB / ID: EMD-13761|
|Title||P. carterae coccolith vesicle with mature crystals|
|Biological species||Chrysotila carterae (eukaryote)|
|Method||electron tomography / cryo EM|
|Authors||Kadan Y / Mahamid J / Gal A / Tollervey F|
|Funding support||European Union, 2 items |
|Citation||Journal: Proc Natl Acad Sci U S A / Year: 2021|
Title: Intracellular nanoscale architecture as a master regulator of calcium carbonate crystallization in marine microalgae.
Authors: Yuval Kadan / Fergus Tollervey / Neta Varsano / Julia Mahamid / Assaf Gal /
Abstract: Unicellular marine microalgae are responsible for one of the largest carbon sinks on Earth. This is in part due to intracellular formation of calcium carbonate scales termed coccoliths. ...Unicellular marine microalgae are responsible for one of the largest carbon sinks on Earth. This is in part due to intracellular formation of calcium carbonate scales termed coccoliths. Traditionally, the influence of changing environmental conditions on this process has been estimated using poorly constrained analogies to crystallization mechanisms in bulk solution, yielding ambiguous predictions. Here, we elucidated the intracellular nanoscale environment of coccolith formation in the model species using cryoelectron tomography. By visualizing cells at various stages of the crystallization process, we reconstructed a timeline of coccolith development. The three-dimensional data portray the native-state structural details of coccolith formation, uncovering the crystallization mechanism, and how it is spatially and temporally controlled. Most strikingly, the developing crystals are only tens of nanometers away from delimiting membranes, resulting in a highly confined volume for crystal growth. We calculate that the number of soluble ions that can be found in such a minute volume at any given time point is less than the number needed to allow the growth of a single atomic layer of the crystal and that the uptake of single protons can markedly affect nominal pH values. In such extreme confinement, the crystallization process is expected to depend primarily on the regulation of ion fluxes by the living cell, and nominal ion concentrations, such as pH, become the result, rather than a driver, of the crystallization process. These findings call for a new perspective on coccolith formation that does not rely exclusively on solution chemistry.
Downloads & links
|File||Download / File: emd_13761.map.gz / Format: CCP4 / Size: 821.3 MB / Type: IMAGE STORED AS SIGNED INTEGER (2 BYTES)|
|Projections & slices|
Images are generated by Spider.
generated in cubic-lattice coordinate
|Voxel size||X=Y=Z: 13.48 Å|
|Symmetry||Space group: 1|
CCP4 map header:
-Entire Pleurochrysis carterae
|Entire||Name: Pleurochrysis carterae / Number of Components: 1|
-Component #1: cellular-component, Pleurochrysis carterae
|Cellular-component||Name: Pleurochrysis carterae / Recombinant expression: No|
|Source||Species: Chrysotila carterae (eukaryote)|
|Specimen||Specimen State: Cell / Method: cryo EM|
|Sample solution||Buffer solution: Sterile artificial seawater, supplemented with an f/2 nutrient recipe|
|Support film||15 mA|
|Vitrification||Instrument: LEICA EM GP / Cryogen Name: ETHANE|
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Imaging||Microscope: FEI TITAN KRIOS|
|Electron gun||Electron Source: FIELD EMISSION GUN / Accelerating Voltage: 300 kV / Electron Dose: 2.24 e/Å2 / Illumination Mode: FLOOD BEAM|
|Lens||Magnification: 42000.0 X (nominal) / Cs: 2.7 mm / Imaging Mode: BRIGHT FIELD / Energy Filter: GIF Quantum LS|
|Specimen Holder||Model: FEI TITAN KRIOS AUTOGRID HOLDER|
|Camera||Detector: GATAN K2 SUMMIT (4k x 4k)|
|Processing||Method: electron tomography / Number of Sections: 49|
|3D reconstruction||Software: IMOD|
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