Detection of Fluorescent Protein Mechanical Switching in Cellulo
<p>This paper describes the detection of fluorescent protein (FP) mechanical switching in genetically encoded Förster resonance energy transfer (FRET)-based tension sensors inside living cells. The authors developed a computational model of FP mechanical switching in the context of FRET-based tension sensors. The model predicts unique signatures in 3-channel FRET imaging data that indicate FP mechanical switching is occurring.</p>
<p>Using the model as a guide, the authors found strong evidence of reversible FP mechanical switching occurring in a synthetic actin-binding tension sensor (ABDTS) in cells. The switching was mainly in the acceptor FP mVenus. They also found weaker but detectable FP mechanical switching occurring in a vinculin tension sensor (VinTS) at focal adhesions in cells. The switching again mainly affected the acceptor mVenus. The FP mechanical switching in VinTS was reduced by disrupting vinculin’s catch bond with actin and eliminated by culturing cells on softer substrates.</p>
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