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TitleMorphological control enables nanometer-scale dissection of cell-cell signaling complexes.
Journal, issue, pagesNat Commun, Vol. 13, Issue 1, Page 7831, Year 2022
Publish dateDec 20, 2022
AuthorsLiam 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 /
PubMed AbstractProtein 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.
External linksNat Commun / PubMed:36539423 / PubMed Central
MethodsEM (tomography)
Structure data

EMDB-15394: Cell-cell contact between two PTK-1 cells
Method: EM (tomography)

Source
  • Potorous tridactylus (long-nosed potoroo)

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