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Mosby, Lewis S.; Hundt, Nikolas; Young, Gavin; Fineberg, Adam; Polin, Marco; Mayor, Satyajit; Kukura, Philipp und Koster, Darius (2020): Myosin II Filament Dynamics in Actin Networks Revealed with Interferometric Scattering Microscopy. In: Biophysical Journal, Bd. 118, Nr. 8: S. 1946-1957

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Abstract

The plasma membrane and the underlying cytoskeletal cortex constitute active platforms for a variety of cellular processes. Recent work has shown that the remodeling acto-myosin network modifies local membrane organization, but the molecular details are only partly understood because of difficulties with experimentally accessing the relevant time and length scales. Here, we use interferometric scattering microscopy to investigate a minimal acto-myosin network linked to a supported lipid bilayer membrane. Using the magnitude of the interferometric contrast, which is proportional to molecular mass, and fast acquisition rates, we detect and image individual membrane-attached actin filaments diffusing within the acto-myosin network and follow individual myosin II filament dynamics. We quantify myosin II filament dwell times and processivity as functions of ATP concentration, providing experimental evidence for the predicted ensemble behavior of myosin head domains. Our results show how decreasing ATP concentrations lead to both increasing dwell times of individual myosin II filaments and a global change from a remodeling to a contractile state of the acto-myosin network.

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