Unraveling the pressure sensitivity of cytoskeletal microtubules

本提案的中心目标是使用高静水压力（HHP）来实现对重要和压力敏感的非共价细胞骨架聚合物的构象和能量景观的整体生物物理描述。在第一个资助期，我们建立了一个基于光谱、x射线散射和显微技术的hhp测量平台，该平台提供了对肌动蛋白聚合和捆绑过程的压力敏感性的深刻见解和详细的机制描述，同时考虑了细胞内可能遇到的因素（即大分子拥挤的影响、渗透物和辅助结合蛋白的存在）。在第二个资助期，我们将重点研究微管装配和动力学的压力摄动，以高时空分辨率研究其热力学、动力学和结构效应。结合我们从微丝压力研究中获得的结果，新的实验将对极端环境条件下生长的生物细胞动力学和运动的抗压机制策略产生新的见解。这些研究对于需要扩展温度-压力过程参数范围的生物技术应用也很有价值。

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.