Sensing and Stiffening of Tubular Membranes by the Syndapin 1 FBAR

Abstract

Bin-Amphiphysin-Rvs (BAR) domains are essential components of the cellular machinery responsible for membrane deformation and were found to be sensitive sensors of membrane curvature. One common feature of BAR domains is a curved shape which correlates with high membrane curvatures often found in cells. We focused on characterizing the FBAR domain of Syndapin-1 in vitro, given its medical relevance to neurological diseases, using tubular membranes pulled out of Giant Unilamellar Vesicles (GUVs). Specifically, GUVs were synthesized using either porcine brain lipid extracts or the more conventional, binary lipid mixtures to systematically test how lipid composition affects curvature sensing activity. Using optical trapping coupled with force-spectroscopy and confocal microscopy, we discovered an inverse relationship between the curvature sensing activity of the FBAR domain and its equilibrium concentration in solution. At high bulk concentrations of protein, we explicitly measured an increase in the tube's persistence length, which can be understood as mechanical stiffening of the tube. Lastly, we used force spectroscopy to accurately test the effect of the protein on membrane relaxation dynamics in real time.

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