This study evaluates the influence of vinculin in closed conformation on the mechanical properties of cells. We demonstrate that MEFvin(-/-) cells transfected with the eGFP-vinculin mutant A50I (talin-binding-deficient-vinculin in a constitutively closed conformation) show 2-fold lower stiffness and focal adhesion density compared to MEFvin(+/+) and MEF(Rescue) cells. MEF(A50I) cells are as stiff as MEFvin(-/-) cells with similar focal adhesion density. Further, 2D traction microscopy indicates that MEF(A50I) and MEFvin(-/-) cells generate 3- to 4-fold less strain energy than MEFvin(+/+) and MEF(Rescue) cells. These results demonstrate that vinculin's mechano-coupling function is dependent on its conformational state.
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Head/tail interaction of vinculin influences cell mechanical behavior.
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Center for Medical Physics and Technology, Biophysics Group, Friedrich-Alexander-University Erlangen-Nuremberg, Henkestrasse 91, 91052 Erlangen, Germany.
This study evaluates the influence of vinculin in closed conformation on the mechanical properties of cells. We demonstrate that MEFvin(-/-) cells transfected with the eGFP-vinculin mutant A50I (talin-binding-deficient-vinculin in a constitutively closed conformation) show 2-fold lower stiffness and focal adhesion density compared to MEFvin(+/+) and MEF(Rescue) cells. MEF(A50I) cells are as stiff as MEFvin(-/-) cells with similar focal adhesion density.
View Article and Find Full Text PDFAnchorage of vinculin to lipid membranes influences cell mechanical properties.
Biophys J
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Center for Medical Physics and Technology, Biophysics Group, Friedrich-Alexander-University of Erlangen-Nuremberg, Erlangen, Germany.
The focal adhesion protein vinculin (1066 residues) can be separated into a 95-kDa head and a 30-kDa tail domain. Vinculin's lipid binding sites localized on the tail, helix 3 (residues 944-978) and the unstructured C-terminal arm (residues 1052-1066, the so-called lipid anchor), influence focal adhesion turnover and are important for cell migration and adhesion. Using magnetic tweezers, we characterized the cell mechanical behavior in mouse embryonic fibroblast (MEF)-vin(-/-) cells transfected with EGFP-linked-vinculin deficient of the lipid anchor (vinDeltaC, residues 1-1051).
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