Chemical mapping of protein networks that sense cellular force

项目摘要/摘要 身体对机械力或机械感觉的感知和反应能力对各种 从调节血压和呼吸等宏观的生理过程到微小的- 在组织发育过程中，细胞迁移、细胞命运决定和细胞排列等规模化过程。尽管 机械感觉对人体健康的重要性，控制不同组织压力感觉的细节 器官仍然未知。机械敏感离子通道称为对拉伸有反应的压电波通道 或压缩质膜，现在已知在机械信号转导中起重要作用。 细胞，而是压电通道在不同的力的巨大动态范围内调整其反应的能力 细胞类型仍然是一个谜。有证据表明，在不同的地区存在不同的压电相互作用网络 细胞类型(例如，附着于细胞骨架、细胞外基质和/或与其他蛋白质的相互作用 辅助亚单位)，能够校准压电式机械换能器。可靠地识别这样的 由于研究离子通道的技术差距，相互作用面临着重大障碍。 这项提案旨在通过开发一种新的基于蛋白质组的质谱学来解决这些差距 绘制感知细胞力的压电蛋白网络图的方法。该项目的成功将为 洞察许多对人类健康和疾病知之甚少的机械调节过程 提供了一种可推广的新技术，用于定义细胞表面的离子通道相互作用网络。这个 该项目的研究培训将在帕塔普蒂安和克拉瓦特实验室的联合指导下进行，网址为 斯克里普斯研究所，两个设备齐全的实验室，拥有离子通道生理学和 化学生物学/蛋白质组质谱学。与主办方定期举行联席会议 和共同赞助商将提供适当的培训环境，学习新技术，促进职业发展 研究金期间的发展目标。

Project Summary/Abstract The body’s ability to sense and respond to mechanical forces, or mechanosensation, is critical to a variety of physiological processes from macroscopic processes like regulation of blood pressure and breathing to small- scale processes like cell migration, cell fate determination, and cell alignment during tissue development. Despite the importance of mechanosensation in human heath, the details governing pressure sensing in different tissues and organs remain unknown. Mechanosensitive ion channels called PIEZO channels that respond to stretching or compression of the plasma membrane are now known to play an important role in mechanical signaling within cells, but the ability of PIEZO channels to tune their response over a vast dynamic range of forces in different cell types is still a mystery. Evidence points to the existence of distinct PIEZO interaction networks in different cell types (e.g., attachments to the cytoskeleton, extracellular matrix, and/or interactions with other protein auxiliary subunits) that enable calibration of PIEZO mechanotransduction. Reliable identification of such interactions has faced significant obstacles due to gaps in technology for studying ion channels. This proposal will aim to address these gaps by developing a new proteomic mass spectrometry-based approach for mapping PIEZO protein networks that sense cellular force. Success of this project will provide insight into many mechanically mediated processes that are poorly understood in human health and disease and provide a generalizable new technique for defining of ion channel interaction networks on the cell surface. The research training for this project will be conducted with joint mentorship in the Patapoutian and Cravatt labs at the Scripps Research Institute, two well-equipped laboratories with expertise in ion channel physiology and chemical biology/proteomic mass spectrometry, respectively. Regularly occurring joint meetings with the sponsor and co-sponsor will provide an adequate training environment to learn new techniques and facilitate career development goals for the duration of the fellowship.