FORCEBIND: Mechanochemical Regulation Of Focal And Fibrillar Adhesion Proteins

FORCEBIND：焦点和纤维粘附蛋白的机械化学调节

• 批准号: EP/Y036085/1
• 负责人: Rafael Tapia-Rojo
• 金额: $ 150.07万
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2024
• 资助国家: 英国
• 起止时间: 2024 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FY036085%2F1
• 关键词: FORCEBIND; Mechanochemical; Regulation; Focal; Fibrillar

Cells sense and adapt to mechanical forces imposed by their environment. Mechanical cues from the extracellular matrix are detected at focal adhesions, where they are converted into biochemical signals to elicit a cellular response. A primary force-transduction mechanism involves the mechanical and chemical regulation of protein-binding interactions. Understanding these processes from a molecular perspective requires evaluating how these proteins interact under force. However, protein biochemistry assays cannot apply force to molecules in bulk, while classic single-molecule techniques are generally limited to using high forces, which precludes the study of mechanosensing proteins given the low forces involved at cell-matrix adhesions. Our objective is to implement novel single-molecule approaches to unravel the molecular mechanisms underpinning mechanotransduction processes at cellular adhesions. To that aim, we will first develop a new single-molecule instrument combining fluorescence with ultra-stable magnetic tweezers, allowing us to capture binding events in proteins under physiologically relevant forces and timescales. This new technology will enable us to examine how key mechanosensing proteins respond conformationally to force and how these forces regulate the binding interactions and post-translational modifications that underpin their function. Specifically, we will determine how phosphorylation regulates protein nanomechanics, using the focal adhesion kinase as a model system, and delve into the elusive mechanosensing function of tensin, the master regulator of fibrillar adhesions. We anticipate that these efforts will unveil how protein dynamics, interactions, and chemistry are finely regulated by mechanical forces, identifying the key molecular mechanisms used by cells to convert mechanical cues into biochemical responses, of critical relevance for multiple biological fields, from development to cancer.

细胞感知并适应其环境施加的机械力。在局灶性粘附处检测到来自细胞外基质的机械提示，并将其转化为生化信号以引起细胞反应。主要的力转导机制涉及蛋白质结合相互作用的机械和化学调节。从分子的角度了解这些过程需要评估这些蛋白在效力下的相互作用。然而，蛋白质生物化学测定不能将力施加到批量分子上，而经典的单分子技术通常仅限于使用高力，这排除了鉴于细胞矩阵粘附涉及的低力的机械传感器蛋白的研究。我们的目标是实施新型的单分子方法，以阐明细胞粘附处的机械转导过程的分子机制。为此，我们将首先开发出一种新的单分子仪器，该仪器将荧光与超稳定的磁镊相结合，从而使我们能够在生理上相关的力和时间表下捕获蛋白质中的结合事件。这项新技术将使我们能够研究关键机械蛋白如何构成对力的响应，以及这些力如何调节其功能支撑其功能的结合相互作用和翻译后修饰。具体而言，我们将使用焦点粘附激酶作为模型系统来确定磷酸化如何调节蛋白质纳米力学，并深入研究原纤维粘附的主要调节剂Tensin的难以捉摸的机械感应函数。我们预计，这些努力将揭示蛋白质动力学，相互作用和化学如何受到机械力的细节调节，从而确定细胞用于将机械提示转化为生化反应的关键分子机制，这是从发育到癌症的多个生物学领域的关键相关性。