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  • Title: Single-molecule dynamics of mechanical coiled-coil unzipping.
    Author: Bornschlögl T, Rief M.
    Journal: Langmuir; 2008 Feb 19; 24(4):1338-42. PubMed ID: 17973511.
    Abstract:
    We use atomic force microscopy (AFM) to mechanically unzip and rezip a double-stranded coiled-coil structure at varying pulling velocities. We find that force-extension traces exhibit hysteresis that grows with increasing pulling velocity. This shows that coiled-coil unzipping and rezipping do not occur in thermal equilibrium on our experimental time scale. We present a nonequilibrium simulation that fully reproduces the hysteresis effects, giving detailed insight into dynamics of coiled-coil folding. Using this model, we find that seed formation is responsible for the hysteresis. The seed consists of four alpha-helical turns on both strands of the coiled coil. To obtain equilibrium information from our nonequilibrium experiments, we used the Crooks fluctuation theorem (CFT) to calculate the equilibrium free energy of folding for all of the different pulling velocities. The paper presented here lays the groundwork for the study of self-assembly properties of many physiologically relevant coiled-coil structures at the single-molecule level.
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