Mechanism of alpha catenin tension sensing

1132116Leckband智力功绩。该项目扩展了该团队最近的创新发现，表明钙粘附素复合体--所有软组织中细胞对细胞连接处的关键蛋白质--能够感知细胞环境的机械特性，并按比例调节细胞功能。一个主要的目标是使用灵敏的力探测器来测试所提出的钙粘蛋白连接处的机械传感机制。一项新的创新将是开发一种基于荧光的细胞内生物传感器，以直接测试机械力是否会在细胞间粘连处诱导假设力传感器的构象变化。该传感器将能够直接可视化活细胞中钙粘附素粘连的动态、受力驱动的变化。这将改变研究细胞间连接的机械转导的能力，并使活细胞中机械转导的空间地图实时可视化。这项多学科的研究涉及敏感的机械测量、探针开发以及活细胞的动态亚细胞成像。这个项目是创新的，因为基于钙粘素的细胞间机械传感及其对更广泛的细胞力学的影响是一个几乎完全未被探索的研究领域。这项研究将产生广泛的影响，因为钙粘素对组织发生、有组织的组织重塑和调节组织屏障至关重要。钙粘附素感知力的能力将极大地影响它们调节细胞功能的方式，并扩大机械力对细胞间通讯的影响。由于钙粘附素对人类健康和组织生物学很重要，这项工作将影响从发育生物学到生物工程和临床的各个领域。其教育影响在于通过伊利诺伊大学的“数学和工程女孩冒险”(游戏)计划和少数族裔暑期研究机会计划(SROP)，在大学预科和本科阶段接触到妇女和少数族裔。这笔赠款将有助于在游戏讲习班和SROP研究项目中实施这项提议中描述的技术，并继续指导妇女和代表性不足的少数群体。

1132116LeckbandIntellectual merit. This program extends recent innovative findings by this team demonstrating that cadherin complexes--critical proteins at cell-to-cell junctions in all soft tissues--sense the mechanical properties of the cell environment and proportionally regulates cell functions. A major objective is to use sensitive force probes to test proposed mechanisms of mechano-sensing at cadherin junctions. A new innovation will be the development of a fluorescence-based intracellular biosensor to directly test whether mechanical force induces conformational changes in a putative force sensor at intercellular adhesions. This sensor will enable direct visualization of dynamic, force-actuated changes at cadherin-based adhesions in live cells. This will transform the ability to investigate mechanotransduction at intercellular junctions, and enable the real-time visualization of spatial maps of mechanotransduction, in a living cell. This multidisciplinary research involves sensitive mechanical measurements, probe development, and dynamic, subcellular imaging of live cells. This program is innovative because cadherin-based, intercellular mechanosensing and its impact on broader cell mechanics is an almost entirely unexplored research area.This research will have broad impact because cadherins are critical for tissue genesis, organized tissue remodeling, and regulating tissue barriers. Cadherins' capacity to sense force would dramatically impact how they regulate cell functions, and expand the impact of mechanical force on cell-cell communication. Because cadherins are important for human health and tissue biology, this work will impact fields ranging from developmental biology to bioengineering and the clinic. The educational impact lies in the outreach to women and minorities at pre-college and undergraduate levels through the "Girls Adventures in Mathematics and Engineering" (GAMES) program and with the Summer Research Opportunity Program for minorities (SROP) at the University of Illinois. This grant will enable the implementation of technologies described in this proposal in GAMES workshops and in SROP research projects, and the continued mentoring women and underrepresented minorities.