Unraveling the pressure sensitivity of cytoskeletal microtubules

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The central goal of this proposal is to use high hydrostatic pressure (HHP) to achieve an overall biophysical description of the conformational and energy landscape of important and pressure-sensitive non-covalent cytoskeletal polymers. In the first funding period we established a HHP-measuring platform based on spectroscopic, X-ray scattering and microscopic techniques that provided deep insights and detailed mechanistic descriptions of the pressure sensitivity of actin polymerization and bundling processes, also considering factors that can be encountered inside the cell (i.e., the effect of macromolecular crowding, the presence of osmolytes and accessory binding proteins). In the second funding period we will focus on the pressure perturbation of microtubule assembly and dynamics by studying its effect thermo-dynamically, kinetically and structurally with high spatial and temporal resolution. Combined with the results obtained from our pressure studies on the microfilaments, the new experiments will yield new insights into the mechanistic strategies for pressure resistance of cellular dynamics and movements found in living beings thriving under extreme environmental conditions. Such studies will also be valuable for biotechnological applications requiring an extended temperature-pressure process parameter range.