- EMDB-15394: Cell-cell contact between two PTK-1 cells -
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Basic information
Entry
Database: EMDB / ID: EMD-15394
Title
Cell-cell contact between two PTK-1 cells
Map data
Raw tomographic reconstruction of a cryo-FIB lamella through a cell-cell contact of two PTK-1 cells. "Contour level" is a placeholder. Use "Auto" button in IMOD.
Journal: Nat Commun / Year: 2022 Title: Morphological control enables nanometer-scale dissection of cell-cell signaling complexes. Authors: Liam P Dow / Guido Gaietta / Yair Kaufman / Mark F Swift / Moara Lemos / Kerry Lane / Matthew Hopcroft / Armel Bezault / Cécile Sauvanet / Niels Volkmann / Beth L Pruitt / Dorit Hanein / Abstract: Protein micropatterning enables robust control of cell positioning on electron-microscopy substrates for cryogenic electron tomography (cryo-ET). However, the combination of regulated cell boundaries ...Protein micropatterning enables robust control of cell positioning on electron-microscopy substrates for cryogenic electron tomography (cryo-ET). However, the combination of regulated cell boundaries and the underlying electron-microscopy substrate (EM-grids) provides a poorly understood microenvironment for cell biology. Because substrate stiffness and morphology affect cellular behavior, we devised protocols to characterize the nanometer-scale details of the protein micropatterns on EM-grids by combining cryo-ET, atomic force microscopy, and scanning electron microscopy. Measuring force displacement characteristics of holey carbon EM-grids, we found that their effective spring constant is similar to physiological values expected from skin tissues. Despite their apparent smoothness at light-microscopy resolution, spatial boundaries of the protein micropatterns are irregular at nanometer scale. Our protein micropatterning workflow provides the means to steer both positioning and morphology of cell doublets to determine nanometer details of punctate adherens junctions. Our workflow serves as the foundation for studying the fundamental structural changes governing cell-cell signaling.
Download / File: emd_15394.map.gz / Format: CCP4 / Size: 928 MB / Type: IMAGE STORED AS SIGNED BYTE
Annotation
Raw tomographic reconstruction of a cryo-FIB lamella through a cell-cell contact of two PTK-1 cells. "Contour level" is a placeholder. Use "Auto" button in IMOD.
Voxel size
X=Y=Z: 8.114 Å
Density
Minimum - Maximum
-128.0 - 127.0
Average (Standard dev.)
6.7312713 (±15.179936)
Symmetry
Space group: 1
Details
EMDB XML:
Map geometry
Axis order
X
Y
Z
Origin
0
0
89
Dimensions
2048
2048
232
Spacing
2048
2048
232
Cell
A: 16617.473 Å / B: 16617.473 Å / C: 1882.4481 Å α=β=γ: 90.0 °
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Supplemental data
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Sample components
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Entire : cell-cell contact
Entire
Name: cell-cell contact
Components
Cell: cell-cell contact
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Supramolecule #1: cell-cell contact
Supramolecule
Name: cell-cell contact / type: cell / ID: 1 / Parent: 0 / Details: cell-cell contact between two PTK-1 cells
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